Pharmacology & Therapeutics 110 (2006) 503 – 524

www.elsevier.com/locate/pharmthera

Associate editor: O. Binah

The peripheral-type benzodiazepine receptor and the cardiovascular system.

Implications for drug development
Leo Veenman, Moshe Gavish *
Rappaport Family Institute for Research in the Medical Sciences, Technion-Israel Institute of Technology, Department of Pharmacology,
Ephron Street, P.O. Box 9649, Bat-Galim, Haifa 31096, Israel

Abstract

Peripheral-type benzodiazepine receptors (PBRs) are abundant in the cardiovascular system. In the cardiovascular lumen, PBRs are present
in platelets, erythrocytes, lymphocytes, and mononuclear cells. In the walls of the cardiovascular system, PBR can be found in the
endothelium, the striated cardiac muscle, the vascular smooth muscles, and the mast cells. The subcellular location of PBR is primarily in
mitochondria. The PBR complex includes the isoquinoline binding protein (IBP), voltage-dependent anion channel (VDAC), and adenine
nucleotide transporter (ANT). Putative endogenous ligands for PBR include protoporphyrin IX, diazepam binding inhibitor (DBI),
triakontatetraneuropeptide (TTN), and phospholipase A2 (PLA2). Classical synthetic ligands for PBR are the isoquinoline 1-(2-chlorophenyl)-
N-methyl-N-(1-methyl-propyl)-3-isoquinolinecarboxamide (PK 11195) and the benzodiazepine 7-chloro-5-(4-chlorophenyl)-1,3-dihydro-1-
methyl-2H-1,4-benzodiazepin-2-one (Ro5 4864). Novel PBR ligands include N,N-di-n-hexyl 2-(4-fluorophenyl)indole-3-acetamide
(FGIN-1-27) and 7-chloro-N,N,5-trimethyl-4-oxo-3-phenyl-3,5-dihydro-4H-pyridazino[4,5-b]indole-1-acetamide (SSR180575), both posses-
sing steroidogenic properties, but while FGIN-1-27 is pro-apoptotic, SSR180575 is anti-apoptotic. Putative PBR functions include regulation
of steroidogenesis, apoptosis, cell proliferation, the mitochondrial membrane potential, the mitochondrial respiratory chain, voltage-dependent
calcium channels, responses to stress, and microglial activation. PBRs in blood vessel walls appear to take part in responses to trauma such
as ischemia. The irreversible PBR antagonist, SSR180575, was found to reduce damage correlated with ischemia. Stress, anxiety disorders,
and neurological disorders, as well as their treatment, can affect PBR levels in blood cells. PBRs in blood cells appear to play roles in
several aspects of the immune response, such as phagocytosis and the secretion of interleukin-2, interleukin-3, and immunoglobulin A (IgA).
Thus, alterations in PBR density in blood cells may have immunological consequences in the affected person. In conclusion, PBR in the
cardiovascular system may represent a new target for drug development.
D 2005 Elsevier Inc. All rights reserved.

Keywords: Peripheral-type benzodiazepine receptor; Cardiac ischemia; Stress; Anxiety; Neurological disorder; Immune response

Abbreviations: 5HT, 5 hydroxytryptamine (serotonin); AHN 086, 1-(2-isothiocyanatoethyl)-7-chloro-1,3-dihydro-5-(4-chloro-phenyl)-2H-1,4-benzodiazepine-2-one

hydrochloride; ANT, adenine nucleotide transporter; B max, maximal binding density; CBR, central-type benzodiazepine receptor(s); CNS, central nervous system;
DAA1097, N-(4-chloro-2-phenoxyphenyl)-N-(2-isopropoxybenzyl)acetamide; DAA1106, N-(2,5-dimethoxybenzyl)-N-(4-fluoro-2-phenoxyphenyl)acetamide; DBI,
diazepam binding inhibitor; DBI-IR, DBI-like immunoreactivity; EGTA, ethylene glycol bis(2-aminoethyl ether)-N,N,NVNV-tetraacetic acid; FGIN-1, 2 aryl-3-
indoleacetamides; FGIN-1-27, N,N-di-n-hexyl 2-(4-fluorophenyl)indole-3-acetamide; Gy, gray; IBP, isoquinoline binding protein; IC50, 50% inhibitory
concentration; IgA, immunoglobulin A; K d, equilibrium dissociation constant; kDa, kiloDalton; K i, equilibrium inhibitory constant; kg, kilogram; LHRH,
luteinizing hormone releasing hormone; MAO, monoamine oxidase; MBq, mega becquerel; mg, milligram; MPTP, mitochondrial permeability transition pore; nM,
nanomolar; PAP7, PBR-associated protein 7; PBR, peripheral-type benzodiazepine receptor(s); PBRs, peripheral-type benzodiazepine receptors; PET, positron
emission tomography; pk10, protein of 10 kDa; PK 11195, 1-(2-chlorophenyl)-N-methyl-N-(1-methyl-propyl)-3-isoquinolinecarboxamide; PLA2, phospholipase
A2; pmol, picomol; p.o., per orally; PRAX-1, PBR-associated protein 1; Ro5 4864, 7-chloro-5-(4-chlorophenyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one;
RT-PCR, reverse transcriptase-polymerase chain reaction; SSR180575, 7-chloro-N,N,5-trimethyl-4-oxo-3-phenyl-3,5-dihydro-4H-pyridazino[4,5-b]indole-1-
acetamide; TP-18, transport protein 18; Trp, tryptophane; TTN, triakontatetraneuropeptide (DBI 17 – 50); VDAC, voltage-dependent anion channel; AM, micromolar

* Corresponding author. Tel.: +972 4 8295275; fax: +972 4 8295271.

E-mail address: mgavish@tx.technion.ac.il (M. Gavish).

0163-7258/$ - see front matter D 2005 Elsevier Inc. All rights reserved.
doi:10.1016/j.pharmthera.2005.09.007
504 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524

Contents

1. Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 504
2. Molecular structure, classical ligands, and functions of peripheral-type benzodiazepine receptors . . . . . . 504
3. Intracellular location and composition of peripheral-type benzodiazepine receptors . . . . . . . . . . . . . 505
4. Endogenous peripheral-type benzodiazepine receptor ligands . . . . . . . . . . . . . . . . . . . . . . . . 505
5. Synthetic peripheral-type benzodiazepine receptor ligands . . . . . . . . . . . . . . . . . . . . . . . . . . 506
6. Tissue specificity of peripheral-type benzodiazepine receptors . . . . . . . . . . . . . . . . . . . . . . . . 508
7. The ontogenic development of peripheral-type benzodiazepine receptor levels throughout the body . . . . 509
8. Peripheral-type benzodiazepine receptors in the heart . . . . . . . . . . . . . . . . . . . . . . . . . . . . 509
9. Peripheral-type benzodiazepine receptors, heart, and stress. . . . . . . . . . . . . . . . . . . . . . . . . . 510
10. Peripheral-type benzodiazepine receptors and ischemia . . . . . . . . . . . . . . . . . . . . . . . . . . . 510
11. Peripheral-type benzodiazepine receptors in blood vessels . . . . . . . . . . . . . . . . . . . . . . . . . . 511
12. Peripheral-type benzodiazepine receptors in human blood cells . . . . . . . . . . . . . . . . . . . . . . . 513
13. Mitochondrial disease and decreases of peripheral-type benzodiazepine receptor density in blood cells . . . 513
14. Modulations of peripheral-type benzodiazepine receptor density in blood cells due to stress
and anxiety disorders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 513
15. Peripheral-type benzodiazepine receptor density of blood cells in correlation with neurological disorders. . 515
16. Peripheral-type benzodiazepine receptor involvement in immune responses of blood cells . . . . . . . . . 516
17. Summary and conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 517
Acknowledgments. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 518
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 518

1. Introduction are also present in glial cells in the brain (Syapin & Skolnick,
1979; Schoemaker et al., 1983). Intracellularly, PBRs are
In contrast to central-type benzodiazepine receptors (CBRs), reported to be located primarily on mitochondrial membranes
which are located primarily in neurons in the central nervous (Anholt et al., 1986; Antkiewicz-Michaluk et al., 1988;
system (CNS), peripheral-type benzodiazepine receptors
(PBRs) were discovered in the 1970s as benzodiazepine TTN DBI
Protoporphyrin-IX PLA2
binding sites outside the CNS (Braestrup & Squires, 1977).
Thereafter, PBR were found to be abundant in the cardiovas-
IBP
cular system of rats (Le Fur et al., 1983a,1983b; Anholt et al., Bcl-2 pk10
1985). PBRs are found throughout the animal kingdom,
VDAC
including insects, mollusks, pisces, amphibians, aves, and creatine- PAP7
kinase
mammals (Peterson et al., 1988; Eshleman & Murray, 1989;
PRAX-1
Lesouhaitier et al., 1996; Snyder & Van Antwerpen, 1998; ANT

Betti et al., 2003). To date, PBRs have not been detected in

reptiles (Bolger et al., 1986; Giannaccini et al., 1993).
Cardiovascular systems of species other than rat were also
shown to express PBR (mice, Hashimoto et al., 1989; Dumont mitochondrial
apoptosis mitochondrial
permeability modulation of
et al., 1999; dog, Gildersleeve et al., 1996; human, Versijpt et respiration voltage dependent
al., 2000). Hence, the widespread expression of PBR through- calcium channels
steroidogenesis
ischemia microglial
out the animal kingdom, including the cardiovascular system, activation immune response
suggests it may have essential functions. cell proliferation

Fig. 1. The PBR complex consists of several IBP molecules typically in

2. Molecular structure, classical ligands, and
functions of peripheral-type benzodiazepine receptors
PBRs are heterotrimers composed of three protein compo-
nents (Fig. 1): an isoquinoline binding protein (IBP; 18 kDa), a
voltage-dependent anion channel (VDAC; 32 kDa); and an
adenine nucleotide transporter (ANT; 30 kDa) (McEnery et al.,
1992). These three components of the PBR show a high degree
of homology between various species (IBP, Gavish et al., 1999;
ANT, Santamaria et al., 2004; VDAC, Graham & Craigen,
2005). Apart from being abundant in peripheral tissues
(Braestrup & Squires, 1977; Verma & Snyder, 1989), PBRs
conjugation with VDAC and ANT. Note that IBP can function without
interactions with VDAC and ANT. Additional molecules, such as pk10, PRAX-
1, and PAP7, can be linked to the PBR. Furthermore, molecules of the Bcl-2
family and creatine kinase can be attached to VDAC and ANT. Endogenous
ligands that bind to PBR include the following: protoporphyrin IX, DBI and its
metabolite TTN, and PLA2. At the functional end, PBR and its ligands have
been shown to be involved in mitochondrial permeability, apoptosis,
steroidogenesis, cell proliferation, mitochondrial respiration, modulation of
voltage-dependent calcium channels, ischemia, microglial activation, and
immune response. Abbreviations: ANT, adenine nucleotide transporter; DBI,
diazepam binding inhibitor; IBP, isoquinoline binding protein; PAP7, PBR-
associated protein 7; PBR, peripheral-type benzodiazepine receptor; pk10,
protein of 10 kDa; PLA2, phospholipase A2; PRAX-1, PBR-associated protein
1; TTN, triakontatetraneuropeptide; VDAC, voltage-dependent anion channel.
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Mukherjee & Das, 1989). Topographic analysis of the receptor
distribution on the mitochondrial membrane has indicated that
several IBP molecules can be associated with one VDAC
molecule (Papadopoulos et al., 1994) and that this complex is
located at the contact sites between the outer and inner
mitochondrial membranes (Culty et al., 1999). In addition, it
appears that the ratio of IBP to VDAC and ANT is tissue and
treatment specific (Golani et al., 2001; Veenman et al., 2002). It
has also been suggested that IBP may be present without
interconnections with VDAC and ANT; that is, free IBP
(Veenman et al., 2002). Structurally, VDAC forms the core of
a protein complex, which in addition to IBP and ANT, may
include several proteins, such as creatine kinase and proteins of
the Bcl-2 family (Fig. 1) (Papadopoulos et al., 1999; Tsujimoto
& Shimizu, 2000; Dolder et al., 2001). Other proteins which are
found to interact with the PBR complex are the following (Fig.
1): a protein of 10 kDa (pk10) (Blahos et al., 1995); PBR-
associated protein 1 (PRAX-1) (Galiegue et al., 1999); and
PBR-associated protein 7 (PAP7) (Li et al., 2001; Liu et al.,
2003). The full name of the associated protein in PAP7 is protein
kinase A with regulatory subunit Ia (Steinberg et al., 1996).
Classical ligands for PBR include 1-(2-chlorophenyl)-N-
methyl-N-(1-methyl-propyl)-3-isoquinolinecarboxamide (PK
11195) and 7-chloro-5-(4-chlorophenyl)-1,3-dihydro-1-meth-
yl-2H-1,4-benzodiazepin-2-one (Ro5 4864) (File & Pellow,
1983; Le Fur et al., 1983a,1983b,1983c; Schoemaker et al.,
1983). The IBP component of the PBR is considered to be
primarily responsible for binding to PK 11195, while Ro5 4864
and other benzodiazepines may bind to all components of the
PBR complex; that is, VDAC, ANT, and IBP (McEnery et al.,
1992; Garnier et al., 1994; Veenman et al., 2002).
A broad spectrum of putative functions have been suggested
for the PBR (Fig. 1), such as regulation of steroid production
(Papadopoulos et al., 1990, 1992, 1997; Weizman et al.,
1997a,1997b; Kelly-Hershkovitz et al., 1998), involvement in
cell growth and differentiation (Wang et al., 1984; Landau et
al., 1998; Carmel et al., 1999), apoptosis (Bono et al., 1999;
Leducq et al., 2003; Veenman et al., 2004; Levin et al., 2005),
regulation of the mitochondrial membrane potential (Leducq et
al., 2003; Galiegue et al., 2003; Chelli et al., 2004), regulation
of the mitochondrial respiratory chain (Hirsch et al., 1989;
Zisterer et al., 1992), effects on the immune and phagocytic
host defense response (Ruff et al., 1985; Bessler et al., 1992,
1997a, 1997b), modulation of voltage-dependent calcium
channels (Bolger et al., 1990; Krueger, 1995), responses to
stress (Karp et al., 1989; Weizman et al., 1994; Avital et al.,
2001), microglial activation related to brain damage (Miya-
zawa et al., 1995; Veenman & Gavish, 2000; Pubill et al., 2001;
Veenman et al., 2002), and cancer cell proliferation (Kelly-
Hershkovitz et al., 1998; Landau et al., 1998; Carmel et al.,
1999; Gavish et al., 1999), including glial tumor cells
(Richfield et al., 1988; Ikezaki et al., 1990; Veenman &
Gavish, 2000; Veenman et al., 2004). It was also found that
PBR may be involved in ischemia (Myers et al., 1991; Chelli et
al., 2001; Faure et al., 2002). Yet the mechanisms whereby
PBRs take part in many of these functions need further
elucidation (Kletsas et al., 2004).
505
3. Intracellular location and composition
of peripheral-type benzodiazepine receptors
Several studies have suggested that PBR may be located in
other locations than mitochondria (for a review, see Woods &
Williams, 1996). Some of the various putative functions of
PBR have been related to its intracellular location and
composition. It has been suggested that mitochondrial PBR
composed of IBP, VDAC, and ANT take part in apoptotic
mechanisms (Veenman et al., 2004; Levin et al., 2005). VDAC
and ANT are the core components of the mitochondrial
permeability transition pore (MPTP), of which extended
opening may lead to mitochondrial swelling and subsequent
release of mitochondrial cytochrome c, followed by the
activation of a caspase cascade leading to apoptosis (Maaser
et al., 2001;Verrier et al., 2003; Chelli et al., 2004; Kunduzova
et al., 2004; Jorda et al., 2005). These studies mentioned above
suggest that IBP may modulate the function of VDAC and
ANT.
Mitochondrial IBP not conjugated with VDAC and ANT
(i.e., free IBP) is considered to be sufficient for PBR function
in steroidogenesis (Lacapere & Papadopoulos, 2003). In
particular, dimers of IBP are shown to possess enhanced
binding capacity to cholesterol and PK 11195, which in turn
leads to increased transport from cytosolic cholesterol into the
mitochondria, determining the speed of formation of pregnen-
olone, the precursor for steroids (Lacapere & Papadopoulos,
2003). On the basis of these observations, several other names
for IBP have been proposed, for example, transport protein 18
(TP-18) (at the meeting: ‘‘Renaming PBR’’, December 18,
2004, Washington, DC). Furthermore, the mitochondrial
location of PBR suggested that PBR may be involved in the
mitochondrial respiratory chain (Hirsch et al., 1989; Zisterer et
al., 1992; Klegeris et al., 2000). Nuclear/perinuclear-located
IBP is considered to play a role in cell division (Hardwick et
al., 1999; Brown et al., 2000).
It has been suggested that the intracellular location of PBR
in human lymphocytes may correlate with its capability to bind
to various endogenous ligands, such as diazepam binding
inhibitor (DBI) and protoporphyrin IX (Berkovich et al., 1993).
It is possible that this capability to bind ligands may be related
to the ratio of IBP to VDAC and ANT in such locations. ANT,
for example, is not found in the cellular plasma membrane
(Belzacq & Brenner, 2003) and VDAC is not found in the
cellular nucleus (Thinnes, 1992; Godlewski et al., 2002).
4. Endogenous peripheral-type
benzodiazepine receptor ligands
Verma and colleagues (Snyder et al., 1987; Verma et al.,
1987; Verma & Snyder, 1988) found that porphyrins can act as
endogenous PBR ligands, while protoporphyrin IX in partic-
ular displayed binding in the nanomolar range (Table 1).
Findings by Mantione et al. (1988) suggested that phospholi-
pase A2 (PLA2) (Naja naja) is also an endogenous PBR ligand
(Table 1). In addition, in 1988, in situ hybridizations in rats
revealed high levels of DBI mRNA in peripheral tissues that
506 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Table 1
Endogenous PBR ligands
PBR ligands Authors
Protoporphyrin IX Snyder et al., 1987; Verma et al., 1987;
Verma & Snyder, 1988
Phospholipase A2 Mantione et al., 1988
Diazepam binding inhibitor Alho et al., 1988
Triakontatetraneuropeptide Slobodyansky et al., 1989
are rich in PBR (Table 1) (Alho et al., 1988). In human
postmortem tissues, a specific radioimmunoassay was used to
describe the distribution of DBI-like immunoreactivity [DBI-
IR(51 – 70)] (Ball et al., 1989). This DBI-IR(51 – 70) was
unevenly distributed throughout the CNS (Ball et al., 1989).
In human peripheral tissues, highest concentrations of DBI-
IR(51 –70) were found in the liver and kidney (Ball et al.,
1989), reminiscent of the distribution of PBR in living humans,
where PBR density is also high in the liver (Versijpt et al.,
2000). Chromatographic analysis revealed several molecular
forms of DBI-IR(51 – 70), the major form being of greater
molecular weight and hydrophobicity than the synthetic
fragment peptide, DBI(51 –70) (Ball et al., 1989). In peripheral
tissues, but not in the CNS, a small peak of immunoreactivity
was indistinguishable from the synthetic peptide. It was
concluded that DBI-IR(51 –70) is widespread, but that tissue
processing may be different (Ball et al., 1989). Bovolin et al.
(1990) reported that DBI showed competitive inhibition of
binding of PK 11195 to the PBR. This may suggest that DBI is
an endogenous ligand for these receptors. Interestingly, in rats,
the highest DBI-like immunoreactivity content was found in
steroid-producing cells, such as the glomerulosa and fasciculata
cells of adrenal cortex and Leydig cells of testis (Bovolin et al.,
1990). In addition, high DBI expression in these tissues was
shown to correlate with high maximal binding density (B max) of
Table 2
Recently developed synthetic PBR ligands
PBR ligands
1-(2-isothiocyanatoethyl)-7-chloro-1,3-dihydro-5-(4-chloro-phenyl)-2H-1,4-benzodiazepine-2-one hydrochloride (AHN 086)
2-Aryl-3-indoleacetamides (FGIN-1-27)
Pyrrolo[2,1-d][1,5]benzothiazepine derivatives
Pyridopyrrolo- and pyrrolobenzoxazepine derivatives
Derivatives of PK 11195
N-(2,5-dimethoxybenzyl)-N-(4-fluoro-2-phenoxyphenyl)acetamide (DAA1106);
N-(4-chloro-2-phenoxyphenyl)-N-(2-isopropoxybenzyl)acetamide (DAA1097) (Ro5 4864 derivatives)
2-Phenylimidazo[1,2-a]pyridine derivatives
2-Arylpyrazolo[1,5-a]pyrimidin-3-yl acetamides
SSR180575 (7-chloro-N,N,5-trimethyl-4-oxo-3-phenyl-3,5-dihydro-4H-pyridazino[4,5-b]indole-1-acetamide)
STPHSTP ( from a group of peptides with the motif STXXXXP)
4-oxo-1-aryl-1,4-dihydro-indeno[1,2-c]pyrazole diethylamide derivative 14a
[11C]VC193M
[11C]VC195M
[11C]VC198M (carboxamide derivatives)
N,N-dialkyl-2-phenylindol-3-ylglyoxylamides
Tetracyclic compounds derived from FGIN-1-27
N-(5-fluoro-2-phenoxyphenyl)-N-(2-[18F]fluoroethyl-5 methoxybenzyl)acetamide;
N-(5-fluoro-2-phenoxyphenyl)-N-(2-[(18)F]fluoromethoxy-d(2)-5-methoxybenzyl)acetamide
Methyl pyropheophorbide-a analogues
2-phenylpyrazolo[1,5a]pyrimidin-3-yl acetamides
PBR (Bovolin et al., 1990). Triakontatetraneuropeptide (TTN,
DBI 17 – 50), a posttranslational product of DBI, was found to
be a specific PBR ligand (Table 1) (Slobodyansky et al., 1989).
These data suggest possible roles for DBI and its posttransla-
tional products as endogenous regulators of PBR functions.
5. Synthetic peripheral-type
benzodiazepine receptor ligands
Given the obvious importance of PBR for various essential
life functions, considerable effort has been invested to develop
novel ligands in addition to the classical synthetic ligands
specific for PBR, PK 11195, and Ro5 4864. Indeed, an
increasing number of novel PBR ligands is being developed
(Table 2). Some of these novel ligands show interesting
properties as outlined below.
In 1986, Lueddens et al. presented 1-(2-isothiocyanatoethyl)-
7-chloro-1,3-dihydro-5-(4-chloro-phenyl)-2H-1,4-benzodiaze-
pine-2-one hydrochloride (AHN 086), a derivative of Ro5 4864
containing an isothiocyanate moiety, which binds irreversibly to
PBR in the kidney, with a 50% inhibitory concentration (IC50)
for [3H]Ro5 4864 of ¨ 1.3 nM. In more detail, [3H]AHN-086
was found to label the VDAC and ANT components of the PBR
(McEnery et al., 1992). Subsequently, this PBR ligand was used
in a series of studies elucidating some aspects of PBR function,
some of which are mentioned in this review.
Subsequently, it was found that the 2 aryl-3-indoleaceta-
mides (FGIN-1) form a class of compounds that potently (in
the low nM range) and selectively bind to PBRs, and increase
mitochondrial steroidogenesis (Romeo et al., 1992). N,N-di-n-
hexyl 2-(4-fluorophenyl)indole-3-acetamide (FGIN-1-27)
crosses the blood –brain barrier, and for this reason this one
FGIN-1 compound was thought to be potent and efficacious
behaviorally. For example, FGIN-1-27 inhibits neophobia in a
Authors
Lueddens et al., 1986
Romeo et al., 1992
Campiani et al., 1996a
Campiani et al., 1996b, 2002
Cappelli et al., 1997, 2003
Okuyama et al., 1999;
Zhang et al., 2003;
Maeda et al., 2004
Trapani et al., 1999
Selleri et al., 2001
Ferzaz et al., 2002
Gazouli et al., 2002
Campagna et al., 2004
Belloli et al., 2004
Primofiore et al., 2004
Okubo et al., 2004
Zhang et al., 2004, 2005
Chen et al., 2005
Selleri et al., 2005
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
manner that is antagonized by PK 11195 (Romeo et al.,
1992). FGIN-1-27 also appears to possess apoptosis-inducing
properties (Maaser et al., 2005). Following the success of
FGIN-1-27, several derivatives from this molecule were
developed. For example, N,N-dialkyl-2-phenylindol-3-yl-
glyoxylamide derivatives III were designed as conformation-
ally constrained analogues of 2-phenylindole-3-acetamides II
(Primofiore et al., 2004). Most of these FGIN-1-27 deriva-
tives showed a high specificity and affinity for PBR, with an
equilibrium inhibitory constant (K i) in the nanomolar to
subnanomolar range (Primofiore et al., 2004). In addition,
some of these FGIN-1-27 derivatives stimulated steroid
biosynthesis in rat C6 glioma cells with a potency similar
to, or higher than, that of classical ligands (Primofiore et al.,
2004). Some newly developed tetracyclic compounds derived
from FGIN-1-27 also showed high affinity for PBR in the nM
range (Okubo et al., 2004).
The synthesis and cardiovascular characterization of a series
of novel pyrrolo[2,1-d][1,5]-benzothiazepine derivatives was
described by Campiani et al. (1996a). Apart from being potent
and selective PBR ligands, some of these compounds also
showed affinity for L-type calcium channels, displaying potent
depressant activity on the myocardial inotropic parameter with
selectivity for cardiac over vascular tissue (Campiani et al.,
1996a). Pyrrolobenzoxazepine derivatives exhibited IC50 and
K i values in the low nanomolar or subnanomolar range, as
measured by the displacement of [3H]PK 11195 binding
(Campiani et al., 1996b). These ligands were also found to
stimulate steroidogenesis (Campiani et al., 1996b). Since the
dimensions of these ligands were known (i.e., their form and
size), these ligands could be used as ‘‘molecular yardsticks’’ to
estimate the form and size of the ‘‘key hole’’ of PBR for its
ligands (Campiani et al., 1996b). Later, other pyridopyrrolo-
and pyrrolobenzoxazepine derivatives were developed, which
bound to PBR from rat brain and testis with picomolar affinity,
representing the most potent ligands that have been identified
to date (Campiani et al., 2002). In addition, they affected
steroidogenesis in MA10 Leydig cells, having similar potency
and effect as PK 11195 (Campiani et al., 2002). Several of
these pyridopyrrolo- and pyrrolobenzoxazepine derivatives
could also be used as molecular yardsticks to probe the spatial
dimension of the lipophilic pocket L4 in the PBR binding site
(Campiani et al., 2002).
Additionally, several conformationally restrained derivatives
of PK 11195 were designed with the aim of estimating the
spatial dimensions of the PBR binding site systematically, and
some of these new ligands also showed enhanced affinity and
steroidogenic activity with respect to reference PK 11195
(Cappelli et al., 1997, 2003).
Several 2-phenylimidazo[1,2-a]pyridineacetamides also
showed high binding affinity and selectivity toward PBRs. In
addition, they markedly increased the levels of neuroactive
steroids in plasma and cerebral cortex (Trapani et al., 1999). It
was also found that N,N-diethyl-(2-arylpyrazolo[1,5-a]pyrimi-
din-3-yl)acetamides typically were highly selective ligands for
PBR, and some of these were found to increase pregnenolone
formation with a potency similar to Ro5 4864 and PK 11195
507
(Selleri et al., 2001). However, the binding affinities of these
compounds were not particularly high (Selleri et al., 2001).
The 4-oxo-1-aryl-1,4-dihydro-indeno[1,2-c]pyrazole diethy-
lamide derivative, named 14a, was also identified as a selective
ligand for PBR, which displayed an IC50 of 124 nM using
[3H]PK 11195 as a radioligand (Campagna et al., 2004). In
addition, it was suggested that two derivatives of the 2-
phenylpyrazolo[1,5a]pyrimidin-3-yl acetamides could be
added to the classical PBR ligands as tools in the investigation
of PBR function, including steroidogenesis (Selleri et al.,
2005).
The Ro5 4864 derivatives, N-(2,5-dimethoxybenzyl)-N-(4-
fluoro-2-phenoxyphenyl)acetamide (DAA1106) and N-(4-
chloro-2-phenoxyphenyl)-N-(2-isopropoxybenzyl)acetamide
(DAA1097), were found to be PBR selective ligands with high
affinity and potent anxiolytic-like properties in laboratory
animals (Okuyama et al., 1999). The Ro5 4864 derivative,
[11C]DAA1106, was developed as a potent and selective
positron emitting radioligand for PBR (Zhang et al., 2003;
Maeda et al., 2004). In another study, three new carboxamide
derivatives were developed and labeled with carbon-11:
[11C]VC193M, [11C]VC195, and [11C]VC198M (Belloli et
al., 2004). The results of this study indicated that [11C]VC195
is a promising candidate for in vivo positron emission
tomography (PET) monitoring of neurodegenerative processes,
with an in vivo behavior that overlaps that of [11C]PK 11195
(Belloli et al., 2004).
One N-(5-fluoro-2-phenoxyphenyl)-N-(2-[18F]fluoroethyl-
5-methoxybenzyl)acetamide derivative appeared to be a useful
PET ligand for PBR and has been used for imaging PBR in
human brain (Zhang et al., 2004). However, in vivo evaluation
in rodents and primates showed that this ligand was unstable
and rapidly metabolized (Zhang et al., 2005). Therefore, a
deuterium-substituted analogue, N-(5-fluoro-2-phenoxyphe-
nyl)-N-(2-[(18)F]fluoromethoxy-d(2)-5-methoxybenzyl)aceta-
mide ([(18)F]5), as a radioligand for PBR was designed (Zhang
et al., 2005). The deuterium substitution decreased the
radioactivity level of [(18)F]5 in the bone of mouse and
showed specific binding to PBR in the rat brain as determined
by ex vivo autoradiography. However, the PET image of
[(18)F]5 for monkey brain showed high radioactivity in the
brain and skull, suggesting a possible species difference
between rodents and primates (Zhang et al., 2005).
Pyropheophorbides and their metal complexes were synthe-
sized to investigate their applications as nonradioactive PBR
labeling compounds and photosensitizers for use in photody-
namic therapy (Chen et al., 2005). They were found to be
localized in mitochondria and showed significant binding to
PBR (Chen et al., 2005). In some cases, their PBR binding
values were similar to that for PK 11195 (Chen et al., 2005).
However, no direct correlation between PK 11195 displace-
ment ability and photosensitizing efficacy of photosensitizers
was observed (Chen et al., 2005).
Ferzaz et al. (2002) first studied the potential neuroprotec-
tive effects of the novel PBR ligand, 7-chloro-N,N,5-trimethyl-
4-oxo-3-phenyl-3,5-dihydro-4H-pyridazino[4,5-b]indole-1-
acetamide (SSR180575). SSR180575 showed high affinity and
508 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
selectivity for the rat and human PBR. SSR180575 also
appeared to promote steroidogenesis, as well as neuronal
survival and repair in axotomy and neuropathy models (Ferzaz
et al., 2002). In addition, SSR180575 appears to possess anti-
apoptotic properties (Leducq et al., 2003). Moreover, binding
to PBR appeared to be irreversible (Vin et al., 2003). These
properties suggested SSR180575 to be a very useful compound
to study PBR functions.
By screening phage display libraries, peptides were identi-
fied that displaced Ro5 4864 (Gazouli et al., 2002). All of the
isolated peptides showed a conserved motif STXXXXP.
Among these peptides, STPHSTP was the most potent in
displacing Ro5 4864 (IC50 = 10 AM). In addition, the peptide
TAT-STPHSTP inhibited Ro5 4864 stimulated steroid produc-
tion in a dose-dependent manner.
6. Tissue specificity of
peripheral-type benzodiazepine receptors
PBR distribution throughout the body has been studied in
various species. For example, in vitro studies showed the rank
order of binding density in rat to be adrenalHkid-
ney¨heart¨testis¨varyHliver¨brain (Awad & Gavish,
1987; Gavish et al., 1999). PBRs in several rat tissues were
also labeled by intravenous injections of [3H]PK 11195 and
[3H]Ro5 4864 (Benavides et al., 1984a). Binding was saturable
in all tissues studied and regional distribution paralleled the in
vitro binding (Benavides et al., 1984a, Awad & Gavish, 1987;
Gavish et al., 1999). The potency order of compounds able to
displace [3H]PK 11195 was similar in vivo and in vitro in these
tissues (i.e., PK 11195 > PK 11211 > Ro5 4864 > diazepam > di-
pyridamole > clonazepam) (Awad & Gavish, 1987; Benavides
et al., 1984a). Similarly, a radioactive-iodinated analog of PK
11195, [123I]-Iodo-PK 11195, used for tissue distribution
studies in rats, indicated a high uptake of radioactivity in
adrenal glands, heart, lung, and kidneys, which was blocked
63 –87% by preadministration of excess unlabeled PK 11195
(Gildersleeve et al., 1996).
Autoradiographic localization of PBR, using [3H]Ro5 4864
in whole-body sections of neonatal rats, showed that PBRs
were most concentrated in the adrenal cortex and the skin
(Anholt et al., 1985). Substantial levels of PBR were also
evident in the heart, the salivary glands, discrete regions of the
kidney, the epithelium of the lung, the nasal and lingual
epithelia, the lining of the pulmonary arteries, the thymus, the
hair follicles of the vibrissae, the tooth buds, and the bone
marrow (Anholt et al., 1985). Considerable binding of [3H]Ro5
4864 was also observed in the brown fat pads, the liver, and the
spleen, but high levels of nonspecific binding precluded
accurate evaluation of the actual specific binding in these
organs (Anholt et al., 1985). Only low levels of [3H]Ro5 4864
binding sites were found in the brain and [3H]Ro5 4864 binding
sites are virtually undetectable in the skeletal muscle, the eye,
the inner ear, and the gastrointestinal tract (Anholt et al., 1985).
In addition to the lining of the pulmonary arteries of
neonatal rats (Anholt et al., 1985), PBRs were also found to be
present in smooth muscles of various organs in rats, including
blood vessels, myometrium, and gastric smooth muscle (Cox et
al., 1991). Thus, apart from demonstrating the presence of PBR
in various organs throughout the body of rats, these studies also
indicate that the rat cardiovascular system contains relatively
high levels of PBR.
In human cerebral cortex, colon, and kidney membranes,
[3H]PK 11195 was found to bind with high affinity (equilib-
rium dissociation constant; K d T 2 nM), displaying a B max of
PBR of 255 T 23, 1633 T 98, and 1908 T 28 fmol/mg protein,
respectively (Awad & Gavish, 1991). Injections of 112 MBq of
the radioactive PBR ligand [123I]iodo-PK 11195 in healthy
volunteers showed the highest binding density of [123I]iodo-PK
11195 to be found in the steroidogenic organs of the ovary
(138.0 AGy/MBq) and in the testis (109.4 AGy/MBq). Also, in
the human heart, binding density for the radioactive PBR
ligand [123I]iodo-PK 11195 was relatively high (31.2 AGy/
MBq) (Versijpt et al., 2000). Similar levels of [123I]iodo-PK
11195 labeling was found in the upper large intestine, lower
large intestine, and liver (39.9, 38.3, 21.9 AGy/MBq, respec-
tively) (Versijpt et al., 2000). Surprisingly, binding density for
the radioactive PBR ligand [123I]iodo-PK 11195 in the adrenal
of humans was relatively low (16.2 AGy/MBq), suggesting that
species differences can occur (Versijpt et al., 2000). In dogs,
relatively high levels of PBR were found in the heart with the
radioactive PBR ligand [123I]iodo-PK 11195 (Gildersleeve et
al., 1996). In mice, an inflow of [123I]iodo-PK 11195 was
observed in the brain, and among peripheral organs, heart
(42.3%), lungs (133.5%), and kidneys (18.4%) showed
relatively high uptake levels (Dumont et al., 1999). Further-
more, after pretreatment with unlabeled PK 11195, there was a
decrease in accumulation in these latter three peripheral organs
in mice, especially in heart (ca. 55%) and lungs (ca. 80%)
(Dumont et al., 1999). The biodistribution of [3H]PK 11195
was also studied in mice (Hashimoto et al., 1989). In particular,
after intravenous injection of [3H]PK 11195 into the mice, high
accumulations of radioactivity were observed in the heart, lung,
spleen, kidney, and adrenal (Hashimoto et al., 1989). The
radioactivity in these organs was significantly decreased by
coadministration with unlabeled PK 11195, which indicated
that PK 11195 specifically binds to the receptors (Hashimoto et
al., 1989). These studies indicate that not only in rat, but also in
other species, PBRs are present in various organs throughout
the body. Furthermore, the cardiovascular system contains
relatively high levels of PBR in various species, including man.
PBR density was also found to be compartmentalized in
various organs, including the heart (Gehlert et al., 1985). These
authors found, using quantitative receptor autoradiography
with [3H]Ro5 4864, that specific [3H]Ro 5 4864 binding sites
were primarily spread throughout the ventricle wall of the rat
heart. In the brain, specific [3H]Ro5 4864 labeling was seen in
the choroid plexus and ependyma cells. Lower levels of
specific binding were seen in areas corresponding to the
glomerular layer of the olfactory bulb. Binding in the kidney
was associated with the ascending limb of the loop of Henle
and the distal convoluted tubule (Gehlert et al., 1985). In more
detail, PBR immunoreactive staining was found along thick
ascending limbs of Henle’s loops, including the macula densa
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
area, along distal tubules, and along collecting ducts (Bribes et
al., 2002). Also in neonatal rats, PBR were restricted to discrete
regions of the kidney (Anholt et al., 1985). In porcine kidney,
PBR was found to be present in epithelial cells of the deep
cortical and outer medulla (Hauet et al., 2002).
Hormonal control of PBR levels in several organs has been
studied. For example, in thyroidectomized rats, compared to
controls, PBR density is decreased in cardiac ventricles but not
in liver tissue (Kragie & Smiehorowski, 1994). Following
surgical castration of adult male Sprague – Dawley rats a
significant decrease in the density of PBR was observed in
Cowper’s glands (71%) and the adrenal (31%), but not in the
heart (Weizman et al., 1992). Administration of testosterone
acetate prevented the castration-induced PBR depletion (Weiz-
man et al., 1992). Studies in female rats, which were
pharmacologically castrated by chronic administration of the
gonadotropin-releasing hormone agonist decapeptyl [triptore-
lin-d-tryptophane (Trp)(6) luteinizing hormone releasing hor-
mone (LHRH)] suggested that hormonal control of PBR ligand
binding density may act via modulations of interactions
between IBP and VDAC (Golani et al., 2001).
PK 11195 and Ro5 4864 were reported to inhibit calcium
channels in heart tissue (Bolger et al., 1990; Campiani et al.,
1996a). Regarding inhibition of calcium channels, both PK
11195 and Ro5 4864 do not display selectivity between cardiac
and vascular tissue (Campiani et al., 1996a). In contrast, one of
the novel PBR ligands derived from pyrrolobenzoxazepine, 7-
acetoxy-6-(p-methoxyphenyl)pyrrolo[2,1-d][1,5]benzothiaze-
pine, displays a clear-cut selectivity for cardiac over vascular
tissue (Campiani et al., 1996a).
Chronic administration of monoamine oxidase (MAO)
inhibitors to mice was associated with alterations in [3H]Ro5
4864 binding in heart, kidney, and liver (Park et al., 1997).
However, with [3H]PK 11195 binding, such changes were only
detectable in the kidney (Park et al., 1997). Since [3H]PK
11195 binds to the IBP component of the PBR complex, it may
be that MAO has little effect on IBP levels in the heart and liver
of mice. Since [3H]Ro5 4864 binds to VDAC and/or ANT, the
changes in [3H]Ro5 4864 binding due to MAO treatment may
be resulting primarily from changes of VDAC and/or ANT
levels in the heart and liver. In the kidney, all three components
of the PBR may be affected by MAO.
The inhomogeneity of PBR ligand binding densities
between and within tissues, potentially including aspects of
its composition from various components, suggests tissue-
specific functions of PBR. Additionally, the differential
distribution of PBR, including its preferential binding to
specific PBR ligands found throughout the compartments of
the cardiovascular system, suggests that PBR may serve
functions specific for individual components of the cardiovas-
cular system.
7. The ontogenic development of peripheral-type
benzodiazepine receptor levels throughout the body
In the rat heart, PBR density as determined with [3H]PK
11195 increases steadily from 170 T 10 fmol/mg protein 3 days
509
before birth until it reaches its maximum level of 2504 T 140
fmol/mg protein 31 days after birth (Fares et al., 1987a).
Similarly, PBR density in the lungs increases from 781 T 48
fmol/mg 1 day before birth until 5928 T 72 fmol/mg protein 31
days after birth. While a marked, steady increase in PBR
density was demonstrated in the rat heart and lungs, the brain
exhibited negligible ontogenetic changes, from 153 T 13 fmol/
mg protein, at the 20th day of gestation till reaching a density
of 184 T 13 fmol/mg protein 31 days after birth (Fares et al.,
1987a). In guinea pig brain, during embryonic life, PBR
density decreases gradually, without major changes after birth,
suggesting species differences regarding ontogenic develop-
ment of PBR in the brain (Daval et al., 1988).
Postnatally, rats aged 1, 2, 12, 18, and 24 months were
assayed for the development of PBR density in various organs
(Mercer et al., 1992). In some organs, including the heart, no
change in PBR density was observed during this time period. A
3-fold increase in PBR density was demonstrated in testis
during maturation, with maximal values appearing at 18
months, followed by a decline at 24 months (Mercer et al.,
1992). Further studies showed that in the rat neocortex, slight
increases in [3H]PK 11195 (from 183 to 340 fmol/mg protein)
and [3H]Ro5 4864 (from 122 to 220 fmol/mg protein) binding
were observed from the first postnatal week until the 8th
postnatal week (Itzhak et al., 1995). This increase in PBR
density was attributed to the presence of PBR in astrocytes
(Itzhak et al., 1995). In the rat adrenal, mitochondrial PBR
density measured with [3H]PK 11195 was found to increase
from 14.5 to 104 pmol/mg protein from postnatal day 9 to
adulthood (Zilz et al., 1999). In young, mature, and senescent
rats, binding of [3H]Ro5 4864 to membranes from rat
hippocampus was found not to be age related (Pedigo et al.,
1981). However, a significant decrease in [3H]Ro5 4864
binding to kidney membranes was demonstrated in senescent
rats (Pedigo et al., 1981).
Thus, it appears that PBR density not only develops
prenatally, but also can change postnatally in various organs
throughout the body. Nonetheless, in several organs, including
the heart, PBR density appears to be stable throughout
postnatal life.
8. Peripheral-type benzodiazepine receptors in the heart
Using [3H]diazepam, Davies and Huston (1981) showed
that PBRs are present in rat and guinea pig hearts. Using
[3H]PK 11195, Le Fur et al. (1983c) reported that binding to
PBR in rat cardiac membranes is saturable, and that these PBRs
show a high affinity for [3H]PK 11195. The order of potency of
[3H]PK 11195 displacing agents in the heart was as follows:
PK 11195 > Ro5 4864 > dipyridamole > diazepam > clonazepam.
The B max in the heart determined with [3H]PK 11195 as a
radioligand was equivalent of the B max determined with
[3H]Ro5 4864 as a radioligand under the same experimental
conditions (Le Fur et al., 1983c). Labeling PBR in the rat heart,
by means of intravenous injection of [3H]PK 11195 and
[3H]Ro5 4864, presented saturable binding that paralleled in
vitro binding (Benavides et al., 1984a). A similar potency order
510 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
of compounds displacing [3H]PK 11195 was found in vivo and
in vitro: PK 11195 > PK 11211 > Ro5 4864 > diazepam > dipyr-
idamole > clonazepam (Awad & Gavish, 1987; Benavides et al.,
1984a).
In both rat and guinea pig heart, PBR binding in the
ventricles appeared to be significantly higher than in the atria, as
determined with [3H]diazepam (Davies & Huston, 1981). PBR
localization was also accomplished using quantitative receptor
autoradiography after labeling slide-mounted tissue sections
with [3H]Ro5 4864 (Gehlert et al., 1985). Specific [3H]Ro5
4864 binding sites were observed throughout the ventricular
wall of the rat heart (Gehlert et al., 1985). In 1989, Bolger et al.
(1989) demonstrated the presence of PBR in guinea pig atrium,
by inhibiting [3H]Ro5 4864 binding with AHN 086. Further,
Bolger et al. (1989) found that AHN 086 and Ro5 4864
enhanced the inotropic responses and blocked the chronotropic
responses of the spontaneously beating guinea pig atrium to the
calcium channel agonist, BAY K 8644. In the isolated working
rat heart model, Ro5 4864 and PK 11195 significantly
decreased the indices of heart contractility (Edoute et al.,
1993). This study showed that aortic flow, stroke work, and total
pressure volume area were also significantly depressed by Ro5
4864 and PK 11195 (Edoute et al., 1993).
PBR densities in various organs subject to hormonal control
appear to be regulated by organotropic hormones (Gavish et al.,
1992). Furthermore, cardiac PBR may be under hormonal
control since densities of PBR were decreased in cardiac
ventricles of thyroidectomized Holtzman adult male rats, as
compared to sham-operated controls (Kragie & Smiehorowski,
1994). The same study showed that PBR affinity for the ligand
was increased in ventricular homogenates from the hypothy-
roid tissues (Kragie & Smiehorowski, 1994).
9. Peripheral-type
benzodiazepine receptors, heart, and stress
Various alterations in PBR density in the heart as a
response to stress have been reported (Gavish et al., 1992).
For example, it has been reported that PBR density in mouse
cardiac ventricles is increased 30 min after acute, maximal
electroshock. At the same time, a significant increase in PBR
density was seen in cerebral cortex, but not in the kidneys
(Basile et al., 1987). In another study, using rats, 80-min
sessions of inescapable tail shock produced reduced [3H]Ro5
4864 binding in cardiac membranes at 2 and 4 hr after stress,
but not after 0 and 1 hr (Drugan et al., 1988). Rats exposed to
6 or 12 hr of stress displayed decreased B max of [3H]PK
11195 in atrial membranes compared to sham-exposed
animals (Salvetti et al., 2000). This was associated with
MPTP opening (Salvetti et al., 2000). Notably, the MPTP
shares its core components, VDAC and ANT, with the PBR
complex, as mentioned in Section 3.
Hence, these studies suggest that PBR density may increase
in heart tissue immediately following stress, but it decreases
after intervals longer than an hour. Further, these changes may
be related with apoptotic mechanisms associated with the
MPTP.
10. Peripheral-type benzodiazepine receptors and ischemia
Rat cardiac mitochondria were used to study the effect of
PBR ligands on apoptosis. Ro5 4864 and AHN 086 induced
mitochondrial Ca2+ release, which was blocked by Mg2+,
whereas the central-type benzodiazepine clonazepam was
ineffective (Moreno-Sanchez et al., 1991). An associated
collapse of the cardiac mitochondrial membrane potential and
mitochondrial swelling, which are preludes to apoptosis, were
also induced by AHN 086 in the presence of Ca2+ (Moreno-
Sanchez et al., 1991). Exposure of de-energized rat cardiac
mitochondria to PBR ligands, such as Ro5 4864, PK 11195,
and diazepam, was shown to produce a dose-dependent and
cyclosporin A-sensitive loss of absorbance, which was
indicative of mitochondrial swelling (Chelli et al., 2001).
Supporting the apoptotic role of PK 11195-induced swelling,
supernatants from mitochondria that had shown permeability
transition caused apoptotic changes in isolated cardiac nuclei
(Chelli et al., 2001). Since apoptosis is one of the processes
leading to cell death during ischemia, PBR are thought to play
a role in ischemia (Chelli et al., 2001; Leducq et al., 2003; Vin
et al., 2003). For example, the irreversible PBR antagonist,
SSR180575 (Section 5; Table 2), greatly reduced the contrac-
tile dysfunction associated with ischemia reperfusion of the
heart. Furthermore, in anesthetized rats, SSR180575 (3– 30
mg/kg po) produced significant reductions in infarct size after
coronary artery occlusion/reperfusion (Leducq et al., 2003). In
humans, PK 11195 (20 mg) did not affect cardiac ischemia
(Drobinski et al., 1989). This may be due to species differences
between rat and man. It may also be possible that SSR180575
is a better candidate to prevent ischemia than PK 11195.
Recently, the involvement of PBR in kidney ischemia has
attracted considerable attention (Faure et al., 2002, 2003; Hauet et
al., 2002; Bono et al., 2004; Kunduzova et al., 2004). Rat
pretreatment with the irreversible PBR antagonist, SSR180575,
decreased postreperfusion oxidative stress and tubular apoptosis
and necrosis in the kidney. This effect was associated with
inhibition of caspase-3 activation, a component of the cascade
leading to apoptosis (Kunduzova et al., 2004). Furthermore,
inhibition of PBR by its antagonist, SSR180575, accelerated the
recovery of normal renal function, as assessed by measurement of
creatine and urea nitrogen plasma levels (Kunduzova et al., 2004).
The role of PBR in relation to forebrain ischemia also
received some attention. Using photochemically induced focal
cortical ischemia as a lesion model in the rat, primary and
secondary lesions were investigated with ex vivo autoradiog-
raphy using [3H]PK 11195 (Myers et al., 1991). Both in the
primary and secondary lesions, due to forebrain ischemia, high
densities of PBR correlated with infiltration of macrophages
(Myers et al., 1991). This may suggest that PBRs are involved
in immune responses during ischemia. Also, the time course of
appearance of PBR around a focally induced ischemic lesion in
frontal cortex of rat brain was established autoradiographically
after intravenous injections of [3H]PK 11195 and [11C]PK
11195 (Cremer et al., 1992). The regional retention of
radioactivity was consistent with immunohistochemical data
for the occurrence of increased numbers of PBR, predomi-
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
nantly in association with macrophages, in forebrain areas
undergoing necrosis (Cremer et al., 1992). Demerle-Pallardy et
al. (1991) studied the accuracy of increases of PBR density as a
marker of neuronal damage induced by transient forebrain
ischemia in the rat (4-vessel occlusion model). Seven days after
forebrain ischemia, a good correlation was found between the
increase of PBR levels (measured using [3H]PK 11195 as a
specific radioligand) in hippocampal, striatal, and cortical
homogenates and the duration of ischemia (Demerle-Pallardy
et al., 1991). The progression of PBR increase was examined
from 3 hr to 14 days of recirculation (Demerle-Pallardy et al.,
1991). After forebrain ischemia in rat, the density of [3H] PK
11195 binding increased rapidly in all strata of the CA1 region
over the first 8 days and then increased more slowly throughout
days 12 – 20 (Conway et al., 1998). Also with rat studies, it
appeared that PBR density after ischemia increased in
microglia in the hippocampus (Stephenson et al., 1995). This
also suggests that PBR may be involved in an immune
response. In the gerbil hippocampus model for reperfusion
after transient forebrain ischemia, a significant increase in PBR
density and PBR mRNA levels between 3 and 7 days of
reperfusion was reported (Rao et al., 2001). At 7 days of
reperfusion, in the hippocampal CA1 (the brain region
manifesting selective neuronal death), PBR immunoreactivity
was also increased significantly (Rao et al., 2001). It was found
for the Mongolian gerbil that benzodiazepines, including
diazepam, protect hippocampal neurons from degeneration
after transient forebrain ischemia (Hall et al., 1997; Schwartz-
Bloom et al., 1998, 2000). However, it was not studied whether
this was due to PBR or CBR.
It is thought that interactions between PBR and voltage-
operated calcium channels in the plasma membrane may play a
role in ischemia. PK 11195 and Ro5 4864 have been found to
possess low but significant inhibitory effect on L-type calcium
channels (Bolger et al., 1990; Campiani et al., 1996a). Ro5 4864
potentiated whereas PK 11195 inhibited, the myocardial ischemia
produced by Bay K 8644 in rat (Bolger et al., 1990). Since PK
11195 and Ro5 4864, however, have been shown to possess
affinity for voltage-operated calcium channels in the plasma
membrane (Campiani et al., 1996a), PK 11195 and Ro5 4864 may
also have direct effects on voltage-operated calcium channels,
apart from their effects via PBR. In a canine model of acute
intoxication with verapamil, a calcium channel blocker, PK 11195
acted as an antidote against some aspects of cardiovascular
depression and arrhythmias (Lheureux et al., 1990). In particular,
sinus activity often was preserved or restored in the animals
treated with PK 11195; however, the hemodynamic alterations
induced by acute verapamil intoxication could not be prevented or
corrected by PK 11195 (Lheureux et al., 1990).
In summary, PBR may play a role in ischemia, including
cardiac ischemia. Regarding the effects of PBR ligands on
ischemia, the irreversible PBR antagonist, SSR180575, reduces
the ischemic damage. It can be hypothesized that SSR180575
antagonism may counteract the increases in PBR density in
tissues affected by ischemia. For example, SSR180575s may
counteract interactions between PBRs and calcium channels.
This may affect immune responses (see Section 16). The
511
studies presented indicate that the classical PBR ligand PK
11195 may exacerbate or prevent damage caused by ischemia,
or have no effect. Preliminary studies in our laboratory dealing
with apoptosis suggest that PK 11195 is a partial agonist for
PBR (unpublished results), which may explain the relatively
inconsistent effects of PK 11195 in regard to ischemia.
11. Peripheral-type
benzodiazepine receptors in blood vessels
PBRs appear to be located in various components of blood
vessels (Table 3, Fig. 2). For example, high levels of PBR are
found in vascular smooth muscle cells of rats (French &
Matlib, 1988; Cox et al., 1991; Bribes et al., 2002). In human
and guinea pig lungs, vascular smooth muscle was not labeled
for PBR (Mak & Barnes, 1990), suggesting an inhomogeneous
distribution of PBR throughout the cardiovascular system, or
species differences. French and Matlib (1988) explored the
existence of a benzodiazepine binding site in rat aortic smooth
muscle membranes employing [3H]Ro 4864 as a radioligand.
The binding site was concentrated in the mitochondrial fraction
enriched with cytochrome c oxidase and semicarbazide-
insensitive MAO (French & Matlib, 1988). In this rat
preparation, [3H]Ro5 4864 bound with high affinity to
membranes of the mitochondrial fraction (French & Matlib,
1988). In particular, the K d, determined from saturation curves
of [3H]Ro5 4864 binding, was 2.8 T 0.7 nM and the B max was
14.0 T 1.2 pmol/mg protein (French & Matlib, 1988). The rank
order of potency (K i) by some PBR ligands, including
benzodiazepines, for inhibition of [3H]Ro5 4864 binding to
isolated membranes was as follows: ( )PK 14067 (6.4 T 0.7
nM) = PK 11195 (6.6 T 0.8 nM) > Ro5 4864 (17.6 T 2.1 nM) >
diazepam (600 T 180 nM) = (+)PK 14068 (530 T 70 nM) >
clonazepam (14,300 T 2100 nM) (French et al., 1989). The
PBR ligand [3H]PK 14105 also bound with high affinity (K d
7.0 T 0.5 nM) and high density (B max = 10.1 T1.5 pmol/mg
Table 3
Location of PBR in components of the cardiovascular system
Components Subcomponents Studies
Heart Ventricle Davies & Huston, 1981; Le Fur et al.,
1983a, 1983b, 1983c; Benavides et al.,
1984a; Gehlert et al., 1985;
Gavish et al., 1992
Blood Endothelial cells Stoebner et al., 1999; Morgan et al., 2004
vessels Smooth muscle French & Matlib, 1988; Mihara &
cells Fujimoto, 1989; Mihara et al., 1990;
Cox et al., 1991; Bribes et al., 2002
Mast cells Taniguchi et al., 1980; Miller 1988;
Davies & Huston, 1981
Blood Erythrocytes Canat et al., 1993; Maeda et al., 1998
Monocytes Canat et al., 1993; Rocca et al., 1993;
Cahard et al., 1994; Maeda et al., 1998
Lymphocytes Canat et al., 1993; Berkovich et al.,
1993; Rocca et al., 1993; Cahard et al.,
1994; Maeda et al., 1998
Platelets Gavish et al., 1986; Mihara &
Fujimoto, 1989; Canat et al., 1993;
Maeda et al., 1998
512 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Ms
SM
En
Lm
Pl
Er
Mn
Fig. 2. In the lumen of the cardiovascular system, PBR are located in platelets
(Pl), erythrocytes (Er), lymphocytes (Lm), and monocytes (Mn). In the walls of
the cardiovascular system, PBRs are present in endothelial cells (En) lining the
lumen, and in smooth muscle cells (SM), as well as in mast cells (Ms) in the
adventitia surrounding the blood vessels. In the heart, PBRs are present in
striated muscles.
protein) to mitochondrial fractions isolated from rat aortic
smooth muscle (Cox et al., 1991). The rank order of potency
of a series of PBR ligands displacing the [3H]PK 14105
binding was as follows: PK 11195 ¨ Ro5 4864 > protoporphy-
rin IX > flunitrazepam > diazepam H clonazepam (Cox et al.,
1991). In rat-isolated aortic rings, PBR ligands (Ro5 4864,
AHN-086, PK 11195, and PK 14105) inhibited in a concen-
tration-dependent and noncompetitive manner, noradrenaline-
induced contractions (Gimeno et al., 1994).
Species differences regarding aortic PBR binding were
found: while [3H]Ro5 4864 bound with high affinity in rat
aorta, this was not the case in porcine aorta (French & Matlib,
1988; Mihara & Fujimoto, 1989). In porcine aortic smooth
muscle membrane, [3H]PK 11195 bound with high affinity to
the membranes (K d = 8.6 T 0.9 nM) (Mihara & Fujimoto, 1989;
Mihara et al., 1990). It was also found that in partially
depolarized muscle strips of rabbit aorta, PK 11195 noncom-
petitively inhibited contractions that were induced by BAY K
8644 (Mestre et al., 1986). This study also showed that
contractions induced by KCl were weakly inhibited by PK
11195 (Mestre et al., 1986).
In the cardiovascular system, PBRs are not only present in
vascular smooth muscle but also in dermal vascular endothelial
cells (Fig. 2) (Stoebner et al., 1999; Morgan et al., 2004). It has
also been suggested that PBR may be present in brain
microvessels, including endothelial cells (Hollingsworth et
al., 1985). In neonatal rats, PBRs were demonstrated in the
lining of the pulmonary arteries (Anholt et al., 1985). Vascular
endothelial cells make up the lining of blood vessels and are
considered to play a role in vasodilatation, mitogen-induced
cell division, and immunologic and inflammatory responses
(Milner et al., 2004). It is interesting in this respect that a role
for PBR in vascular inflammatory responses has been
suggested, in particular in vascular permeability caused by
carrageenin (Lazzarini et al., 2001). This also may have
implications for the role of PBR in inflammatory processes
related to ischemia (Faure et al., 2002). It has been reported that
intravenously administered diazepam, a mixed ligand (i.e.,
binding to both CBR and PBR), causes marked inflammation
with edema and intramural polymorphonuclear cell infiltration
in vascular tissue of rats, suggesting that PBR in the blood
vessels indeed may play a role in inflammation (Graham et al.,
1977). It was also found that selected benzodiazepine
analogues possessing platelet-activating factor antagonist
activity could stimulate the production of tissue-type plasmin-
ogen activator by endothelial cells in vitro (Kooistra et al.,
1993). Plasminogen activators play a role in tissue repair-
associated angiogenesis (Van Hinsbergh et al., 1997). In
particular, tissue-type plasminogen activator is stored in
endothelial cells and can be released acutely into the vessel
lumen upon stimulation of the endothelium to activate
fibrinolysis and to prevent fibrin deposition (Van Hinsbergh
et al., 1997). However, it is not clear whether or not PBRs were
involved in this process (Kooistra et al., 1993).
PBRs are also present in mast cells, which are distributed in
many organs and are located near walls of small blood vessels
(Fig. 2) (Taniguchi et al., 1980; Miller et al., 1988; Davies &
Huston, 1981). Mast cells, which are considered to have
originated from monocytes (Yong, 1997), can produce a host of
biologically active substances, the most notable being serotonin
(5 hydroxytryptamine; 5HT), dopamine, heparin, tryptase, and
chymase. These substances may be released in response to
immunological and neural stimuli (Yong, 1997). Mast cells are
considered to be important for immune responses to pathogens
(Marshall, 2004), and they have also been implicated in the
regulation of thrombosis and in inflammatory and cardiovas-
cular disease processes, such as the development and progres-
sion of atherosclerosis, as well as in neoplastic conditions
(Yong, 1997; Wojta et al., 2003 – 2004). PBRs appear to be
implicated in calcium uptake and serotonin release of mast cells
(Suzuki-Nishimura et al., 1989). In particular, Ro5 4864,
diazepam, and chlordiazepoxide inhibited the concanavalin A-
induced [14C]serotonin release from rat mast cells as well as the
concanavalin A-induced 45Ca uptake (Suzuki-Nishimura et al.,
1989). The potencies of benzodiazepines to inhibit concanav-
alin A-induced serotonin release and 45Ca uptake were
correlated with their binding affinities to PBR (Suzuki-
Nishimura et al., 1989). These results suggest that inhibition
of serotonin release by benzodiazepines in mast cells activated
by concanavalin A is mainly due to inhibition of calcium
channels, which may be controlled by the PBR. Serotonin
released by mast cells may induce elevated Ca levels in
vascular smooth muscle (Weintraub et al., 1994). Serotonin
also has been reported to increase blood – brain barrier
permeability in some studies (Abbott, 2000). Where barrier
opening was seen, there was evidence for activation of 5HT-2
receptors and a calcium-dependent permeability increase
(Abbott, 2000). Serotonin released from mast cells after tissue
injury is known to exert algesic and analgesic effects
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
depending on the site of action and the receptor subtype
(Sommer, 2004).
Thus, benzodiazepines may interact with PBR in mast cells
to inhibit calcium channels and, as a consequence, reduce
serotonin release of mast cells. This reduction in serotonin
release, in turn, may decrease vascular smooth muscle
contractions via reduction of Ca2+ levels in vascular smooth
muscle. Furthermore, reduction in serotonin release may reduce
blood – brain permeability and influence pain levels.
In summary, PBRs appear to be present in endothelial cells,
smooth muscle cells, and mast cells of the vascular wall (Fig. 2,
Table 3). As in heart muscle, PBR located in vascular smooth
muscles may play a role in contractility. The studies presented
above suggest that PBR in endothelial cells and mast cells
possibly are involved in responses to trauma, including
inflammation.
12. Peripheral-type
benzodiazepine receptors in human blood cells
Apart from the tissues mentioned above, including compo-
nents of the vessel walls, PBRs were found to be present in
several human blood cell subpopulations, including erythro-
cytes, platelets, monocytes, and lymphocytes (Fig. 2, Table 3)
(Alexander et al., 1992; Canat et al., 1993; Maeda et al., 1998).
Interestingly in this respect, high levels of PBR density were
found in bone marrow of neonatal rats, a site for blood cell
formation (Anholt et al., 1985). Involvement of PBR in
erythroid differentiation of the human myeloid leukemia
K562 cells has been demonstrated (Koiso et al., 2000).
Pharmacological characteristics of PBR in blood cells were
established by PBR binding studies and by quantitative reverse
transcriptase-polymerase chain reaction (RT-PCR) to assay
PBR mRNA expression (Canat et al., 1993). These data
indicated the following: (1) PBR binding is saturable in the
various types of blood cells; (2) the rank order of PBR density
in blood cells is as follows: monocytes = polymorphonuclear
neutrophils > lymphocytes H platelets > erythrocytes; and (3)
PBRs in blood cells appear to be transcriptionally regulated
since mRNA levels are roughly correlated with PBR density
(Canat et al., 1993). Studies of the localization of PBR in two
human leukocyte populations, T4-lymphocytes and monocytes,
showed that there are PBR containing and non-PBR containing
mitochondria (Cahard et al., 1994). Furthermore, 50% of the
PBRs are localized in cell compartments devoid of visible
mitochondria (Cahard et al., 1994). This suggests that some
populations of PBR in immune cells may have physiologic
functions other than associated with mitochondria. One study
suggested that PBRs located on the cell plasma membrane of
lymphocytes may bind to DBI, but not to protoporphyrin IX,
while PBRs located on the mitochondria of lymphocytes may
bind to both protoporphyrin IX and DBI (Berkovich et al.,
1993). Interestingly, PBR and its endogenous ligand, DBI,
appeared to be colocalized in all major lymphocyte sets and
subsets, as well as in monocytes (Rocca et al., 1993).
Furthermore, a significant correlation was observed between
the density of PBR in a given cell type and its abundance of
513
DBI-like immunoreactivity (Rocca et al., 1993). In particular,
for both PBR and DBI-like immunoreactivity, the cell
distribution was as follows: monocytes > B cells and large
granular lymphocytes > T-cells. The presence of PBR and its
endogenous ligand DBI in the various types of blood cells
suggests that PBR may be involved in their cellular functions.
Interestingly, species differences between PBR affinities of
blood cells were observed. In particular, when [3H]PK 11195
binding was examined in porcine, human, and rat platelets, Ro5
4864 inhibited the binding in porcine and human platelets one
order of magnitude less potently than that in rat platelets
(Mihara & Fujimoto, 1989). Using intact platelets, Benavides
et al. (1984b) showed that the B max of [3H]PK 11195 of human
and rat platelets is very high (B max > 108 pmol/mg protein).
Benavides et al. (1984b) also showed that, in contrast to
[3H]PK 11195 binding, [3H]Ro5 4864 binding to membranes is
not without problems (e.g., Ro5 4864 affinity is greatly
diminished due to lysis in comparison to intact cells), while
this is not the case for PK 11195. Benavides et al. (1984b)
stated that for this reason [3H]Ro5 4864 binding can only be
demonstrated in intact cells.
13. Mitochondrial disease
and decreases of peripheral-type
benzodiazepine receptor density in blood cells
Since PBRs are found on the outer mitochondrial membrane,
the possibility of PBR involvement in mitochondrial disorders
was investigated. Using [3H]PK 11195 as a ligand, PBR binding
characteristics were assayed in platelets from patients affected
by mitochondrial disorders with large-scale mitochondrial DNA
deletions and reduced cytochrome c oxidase activity (Martini et
al., 2002). PBR densities in platelets of mitochondrial disorders
patients compared to controls were relatively low (B max
2387.0 T 305.6 vs. 4889.0 T 357.8 fmol/mg protein, P < 0.05),
whereas the K d values in patients were higher than in controls
(13.18 T 2.06 vs. 5.63 T 0.46 nM, P < 0.05). Thus, in mitochon-
drial disorders, alterations in PBR characteristics appear to be
related to the mitochondrial dysfunction.
14. Modulations of
peripheral-type benzodiazepine receptor density
in blood cells due to stress and anxiety disorders
It is well known that stress affects hormonal levels, and
several studies have suggested that PBR ligand binding density
may be under hormonal control (Fares et al., 1987b, 1988,
1989; Bar-Ami et al., 1989; Weizman et al., 1992; Golani et al.,
2001). Furthermore, it was shown that high dose steroid
treatment of multiple sclerosis led to an increase in PBR on
blood mononuclear cells (Ferrero et al., 1992). Thus, it was
considered possible that stress also affects PBR levels. Studies
assaying PBR levels in blood cells during and following
stressful situations are presented in Table 4. Indeed, assaying
students immediately after an exam showed that increased PBR
density in platelets accompanies stress as compared to the
controls (Karp et al., 1989). Furthermore, stress of war was also
514 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Table 4
Blood cell PBR involvement in stress, anxiety, and related disorders
Condition Type of blood cell
Acute stress Platelets
Lymphocytes
Repeated stress Platelets
Posttraumatic stress disorder Platelets
Generalized anxiety disorder Platelets
Platelets
Lymphocytes
Mononuclear cells
Monocytes
Diazepam-treated generalized anxiety disorder Platelets
2V-chloro-N-desmethyl-diazepam-treated Mononuclear cells
generalized anxiety disorder
Adult separation anxiety Platelets
Generalized social phobia Platelets
Obsessive – compulsive disorder Lymphocytes
Major depression Platelets
Electroconvulsive therapy-treated major depression Platelets
Suicidal Platelets
found to increase PBR density in platelets (Weizman et al.,
1994). It was suggested that PBR density in platelets can be
used as a promising biological marker of stressful conditions
(Nakamura et al., 2002). A more recent study also showed that
PBR density in blood platelets was significantly increased
following PhD examination (Droogleever Fortuyn et al., 2004).
In addition, plasma levels of cortisol and allopregnanolone
were also found to be elevated in this study (Droogleever
Fortuyn et al., 2004). These studies suggest that acute stress
may increase PBR density on platelets in correlation with
plasma steroid levels. Another study found that following
academic examinations, PBR mRNA levels in lymphocytes
were increased in some students and decreased in others
(Nudmamud et al., 2000). It was suggested that these differing
effects may have depended on the confidence of the students
following the examinations (Nudmamud et al., 2000). Thus, not
only platelets but also other blood cells may show elevated
PBR density as a consequence of acute stress. The average PBR
levels correlated positively with cortisol and prolactin levels for
the two groups of different students, but this was not so for
individual students (Nudmamud et al., 2000). This suggests that
while average PBR levels are correlated positively with serum
steroid levels, this relation is not a causal one.
Not only the effects of acute stress on PBR blood cell
were studied, as discussed above, but also the effects of repeated
chronic stress and posttraumatic stress disorder on blood cell
PBR levels (Table 4). [3H]PK 11195 binding to platelet
membranes was reduced immediately after actual repeated
parachute jumps (26%; P < 0.05) toward the end of parachute
training of soldiers (Dar et al., 1991). Decreased platelet PBR
density ( 62%; P < 0.001) was also observed in the posttraumatic
stress disorder patients compared to controls (Gavish et al., 1996).
The decreased levels of PBR levels in platelets correlated
with repeated stress and posttraumatic stress disorder suggested
that anxiety may affect PBR levels in blood cells. Studies
dealing with this question are listed in Table 4. In healthy
people, it was found that PBR on platelets are significantly and
PBR levels Study
Up-regulated Karp et al., 1989; Weizman et al., 1994;
Droogleever Fortuyn et al., 2004
Variable Nudmamud et al., 2000
Up-regulated Dar et al., 1991; Gavish et al., 1996
Up-regulated Gavish et al., 1996
Down-regulated Weizman et al., 1987a
Up-regulated Chiu et al., 2001
Down-regulated Nudmamud et al., 2000
Down-regulated Rocca et al., 1998
Down-regulated Sacerdote et al., 1999
Up-regulated Weizman et al., 1987a
Up-regulated Rocca et al., 1998
Down-regulated Pini et al., 2005
Down-regulated Johnson et al., 1998
Down-regulated Rocca et al., 2000
No effect Weizman et al., 1995
Down-regulated Weizman et al., 1996
Down-regulated Soreni et al., 1999
positively correlated with trait anxiety scales of the State-Trait
Anxiety Inventory, but not with scores of state anxiety
(Nakamura et al., 2002). It was therefore suggested that the
density of platelet PBR is highly associated with these
personality traits for anxiety tolerance (Nakamura et al.,
2002). Reduced [3H]PK 11195 binding capacity to platelet
membranes was observed in anxious patients in comparison to
age- and sex-matched normal controls (Weizman et al., 1987a).
Another group, using the PBR ligand [3H]Ro5 4864 instead of
the commonly used [3H]PK 11195, detected enhanced PBR
binding in platelets of patients diagnosed with generalized
anxiety disorder (Chiu et al., 2001). Note that in human
platelets, Ro5 4864 binds one order of magnitude less potently
than in rat platelets, as mentioned above (Mihara & Fujimoto,
1989). Other studies suggested that PBR levels in platelets of
patients with generalized social phobia were decreased (Johnson
et al., 1998). Patients with adult separation anxiety symptoms
were also found to have significantly lower densities of PBR on
platelets as determined with [3H]PK 11195 (Pini et al., 2005).
Also adolescent suicidal patients showed a significant decrease
in platelet PBR density compared to the nonsuicidal control
groups (Soreni et al., 1999). Thus, platelet PBR levels are
typically reduced in patients suffering from various types of
anxiety disorders, and also in suicidal patients (Table 4).
In peripheral blood mononuclear cells of generalized
anxiety disorder patients, it was found that PBR density was
decreased by 45% (as assayed with [3H]PK 11195), while PBR
mRNA levels were decreased by 70% (determined by RT-
PCR), as compared to untreated healthy controls (Rocca et al.,
1998). In addition, in a group of clinically anxious patients the
average level of PBR mRNA in lymphocytes was also found to
be reduced compared to the control group (Nudmamud et al.,
2000). In obsessive –compulsive disorder patients, the relative
content of PBR mRNA in lymphocytes was examined by RT-
PCR. PBR mRNA was significantly decreased only in chronic
obsessive –compulsive disorder patients as compared to con-
trols, whereas no significant changes were observed in episodic
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524 515

obsessive – compulsive disorder patients (Table 4) (Rocca et al.,
2000).
It was also found that in patients suffering from generalized
anxiety, benzodiazepine-induced chemotaxis of monocytes is
completely abolished, while the response to the control
chemoattractant formyl-leu-met-phe is still maintained. The
chemotaxis responses are not restored after pharmacological
treatment of the pathology (Sacerdote et al., 1999). The
decreased chemotactic response could be linked to a decreased
number of PBR present on monocytes of generalized anxiety
disorder patients.
Treatments of various disorders can also have effects on
blood cell PBR levels. Four weeks of diazepam treatment of
anxious patients induced elevation of PBR density of
platelets (Weizman et al., 1987a). Peripheral blood mononu-
clear cells of generalized anxiety disorder patients treated
with 2V-chloro-N-desmethyl-diazepam also showed increases
of PBR density and PBR mRNA levels back to the levels of
the untreated healthy controls (Rocca et al., 1998). Further-
more, this suggests that the decrease of PBR density in
peripheral mononuclear cells of generalized anxiety disorder
patients is due to a change at the transcriptional level (Rocca
et al., 1998). It was found that major depression, in contrast
to stress and some anxiety disorders, did not appear to be
associated with PBR density of platelets (Table 4) (Weizman
et al., 1995). Nonetheless, electroconvulsive therapy down-
regulates platelet PBR in medication-resistant major de-
pressed patients (Table 4) (Weizman et al., 1996). Similarly,
binding studies with [3H]PK 11195 on human platelets
showed that chronic consumption of alcohol did not alter
PBR density of platelets, as compared to nonalcoholics (Karp
et al., 1991). Treatment for 3 weeks with disulfiram of
chronic alcoholics, however, resulted in a significant increase
in the PBR density of platelets (Karp et al., 1991). In
humans, the effects of gonadal steroids on lymphocytic PBR
density were studied in relation the premenstrual syndrome
(Daly et al., 2001). In this model, however, PBR densities
were not altered in women with premenstrual syndrome and
were not changed by selective gonadal steroid administration.
Thus, drugs or other treatments can affect PBR levels in
blood cells.
In summary, these studies of PBR ligand binding in blood
cells suggest that PBR density in platelets does increase with
acute stress (Table 4). With anxiety disorders (as well as after
repeated stress), PBR levels in platelets, lymphocytes, and
monocytes typically are decreased in comparison to controls
(Table 4). Decreased PBR density was also reported in
lymphocytes of patients with obsessive –compulsive disorder
and in platelets of patients at suicidal risk (Table 4). Regarding
the overall picture, it may be considered that PBR density in
blood cells can be increased due to acute stress, but decreased
under conditions of chronic stress such as occur with anxiety
disorders. It may be so that the decreased levels of PBR in
blood cells in anxiety patients may be correlated with reduced
immune responses. Treatment of disorders can modulate PBR
density in human blood cells (Table 4), either to return to
normal levels, or to deviate from the norm. This could have
consequences for the immune system.
15. Peripheral-type benzodiazepine receptor density
of blood cells in correlation with neurological disorders
Since PBRs of blood cells are affected by chronic disorders
such as anxiety disorders, posttraumatic stress disorder, and
major depression, which may include impairments of brain
function, the possibility that PBR may be also be affected by
neurological diseases should be considered. For example, PBR
may have a role in epilepsy and in mediating antiepileptic drug
effects (Veenman & Gavish, 2000; Veenman et al., 2002).
Studies dealing with neurological disorders and blood cell PBR
levels are listed in Table 5. Untreated epileptic patients showed
a significant decrease of 17.1% in PBR density in platelets, as
compared to healthy controls (Weizman et al., 1987b). In
contrast, an increase of 50.3% was observed in carbamazepine-
treated patients as compared to their pretreatment values
(Weizman et al., 1987b). Possible modifications of PBR
density and levels of its endogenous ligand, DBI, in
lymphocytes of epileptic patients treated with various drugs
were also studied. PBR levels were 50 – 80% higher in patients
treated with carbamazepine, phenobarbital, and valproic acid
than in controls and untreated epileptics (Ferrarese et al., 1996).
DBI levels were significantly increased in the lymphocytes of

Table 5
Blood cell PBR involvement in neurological disorders
Condition Drug treatment Type of blood cell PBR levels Study
Epilepsy No Platelets Down-regulated Weizman et al., 1987b
Carbamazepine Platelets Up-regulated Weizman et al., 1987b
No Lymphocytes No effect Ferrarese et al., 1996, 1997
Drug resistant Lymphocytes Down-regulated Ferrarese et al., 1997
Carbamazepine, phenobarbital, valproic acid Lymphocytes Up-regulated Ferrarese et al., 1996
Parkinson’s disease No Platelets Down-regulated Bonuccelli et al., 1991
l-DOPA plus dopamine agonists Lymphocytes Up-regulated Blandini et al., 2004
Schizophrenia No Platelets No effect Gavish et al., 1986; Weizman et al., 1986
‘‘Acute’’ neuroleptics Platelets No effect Tanne et al., 1987
Chronic neuroleptics Platelets Down-regulated Gavish et al., 1986
Schizophrenia ‘‘aggressive’’ N/A Platelets Down-regulated Ritsner et al., 2003
Alzheimer’s disease No Platelets Down-regulated Bongioanni et al., 1996, 1997
No Platelets Up-regulated Bidder et al., 1990
516 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
untreated patients, compared to healthy controls (Ferrarese et
al., 1996). Anticonvulsant therapy led to slight increases in
DBI levels in lymphocytes of these epileptic patients (Ferrarese
et al., 1996). A relatively low PBR density was observed in
lymphocytes of drug-resistant epileptic patients, compared to
newly diagnosed patients and to normal age-related controls
(Ferrarese et al., 1997). The latter suggests that lymphocyte
PBR binding may relate to different subgroups of epileptic
patients. The decrease in platelet PBR binding in epileptic
patients was not considered to depend on patient subgroups
(Weizman et al., 1987b).
Platelet PBR density was also found to be decreased in
untreated Parkinson’s disease patients (Bonuccelli et al., 1991).
Changes in the regulatory mechanisms of apoptosis in
Parkinson’s disease patients were detected in lymphocytes
(Blandini et al., 2004). In particular, patients treated with l-
DOPA in combination with dopamine agonists showed
increased PBR density in lymphocytes, but low levels of Bcl-
2, compared to untreated Parkinson’s disease patients and
healthy volunteers. Increased caspase-3 activity and Cu/Zn
superoxide dismutase levels were also found in these patients
(Blandini et al., 2004). The low levels of Bcl-2 and increased
caspase-3 activity in these l-DOPA/dopamine agonists treated
dopamine patients may be due to the regulatory function of IBP
on VDAC and ANT in regard to apoptosis, as postulated in
Section 3.
In platelets of schizophrenics maintained on chronic
neuroleptic treatment, a decrease of 30% was observed in
PBR density, assayed with [3H]PK 11195, as compared to
platelets of untreated schizophrenics and healthy volunteers
(Gavish et al., 1986; Weizman et al., 1986). In contrast to
chronic treatment, only 28 days of neuroleptic treatment of
schizophrenic patients did not affect the binding characteristics
of [3H]PK 11195 to platelets (Tanne et al., 1987). Furthermore,
it was found that currently aggressive schizophrenic patients
displayed lower platelet PBR density, compared to homicidal
and nonaggressive schizophrenia patients and healthy controls
(Ritsner et al., 2003). These relatively low platelet PBR levels
in aggressive schizophrenia patients were associated with
higher scores for overt aggression, hostility, and anxiety, but
independent of illness subtype, homicidal and suicidal attempt
history, distress level, and type of antipsychotic treatment
(Ritsner et al., 2003). Thus, this subgroup of schizophrenia
patients may be characterized by low platelet PBR binding
levels, possibly related to their relatively high anxiety levels.
This is in contrast to other schizophrenic patients, which do not
show alterations in platelet PBR levels.
Using [3H]PK 11195, a reduction in PBR B max values of
platelets was also observed in demented patients (Bongioanni
et al., 1996). In another study, assaying platelet and lympho-
cyte PBR binding in patients with Alzheimer’s disease and
healthy controls, a reduced number of PBR in patients’
platelets and lymphocytes was found (Bongioanni et al.,
1997). In one study regarding Alzheimer disease, PBR density
on platelets was increased (Bidder et al., 1990). Discrepancies
between different studies possibly may by due to lack of
homogeneity between Alzheimer’s disease patient groups.
Further, studies on epilepsy and schizophrenia (see above)
suggested that subgroups of neurological patients may display
specific changes in blood cell PBR density, which differ from
what is observed in the main group of the neurological disorder
in question. Possibly, factors apart from Alzheimer’s disease
play a more important role in the PBR density of blood cells. It
may be that in one experimental situation the patients are
stressed, while in another situation the patients are anxious.
Finally, it has been hypothesized that PBR density on
peripheral blood cells may be correlated with an impaired
immune response in Alzheimer’s disease patients (Bongioanni
et al., 1997).
In summary (see Table 5), the limited number of
experiments with neurologic patients makes it difficult to
draw definite conclusions about the effects of neurologic
orders on PBR densities in blood cells of epileptic,
Parkinson’s disease, schizophrenic, and Alzheimer’s disease.
Drug treatment appears to enhance PBR density in platelets of
epileptic patients as well as in lymphocytes of epileptic and
Parkinson’s disease patients (Table 5). In schizophrenic
patients, neuroleptic treatment down-regulated platelet PBR
density. As mentioned above, also drug treatment of anxiety
patients and chronic alcoholics, and electroconvulsive therapy
for major depression, can also modulate PBR density on
platelets (Table 4). Presently, it seems unpredictable whether a
treatment will up- or down-regulate PBR levels on blood
cells. Nonetheless, the effects of treatments on PBR levels of
blood cells may be relevant for these components of the
immune system.
16. Peripheral-type benzodiazepine receptor
involvement in immune responses of blood cells
In mice, benzodiazepines were found to affect polymorpho-
nuclear cell chemotaxis and phagocytosis, general immunity,
and survival of infections (Galdiero et al., 1995). Several
studies also examined potential PBR involvement in immune
responses of human blood cells, as listed in Table 6.
Immunomodulatory effects of ligands active at PBR were
examined in human peripheral blood mononuclear cells
(Bessler et al., 1992). Ro5 4864, PK 11195, and diazepam
suppressed phytohemagglutinin and concanavalin A induced
proliferation of peripheral blood mononuclear cells. All three
ligands inhibited interleukin-3-like activity secretion, while the
production of interleukin-2 was inhibited only by Ro5 4864
and diazepam. The selective CBR ligand clonazepam did not
affect the cellular immune functions examined (Bessler et al.,
1992). The in vitro immune-suppressive activity of PBR and
mixed PBR/CBR, but not CBR ligands, indicates that this
immunosuppression is due to PBR (Bessler et al., 1992).
Furthermore, since PK 11195 did not affect interleukin-2
production, it appears that the IBP component of the PBR
complex is not involved in this specific component of the
immune response. Nonetheless, it was found that both the
specific PBR ligands Ro5 4864 and PK 11195, inhibited in a
dose-dependent manner (in the micromolar range) the sponta-
neous secretion of immunoglobulin A (IgA) by human
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Table 6
Effect of PBR ligands on immune responses of human blood cells
Cell type Immune response
Mononuclear cells Proliferation, interleukin-2 and -3 secretion,
and IgA secretion
Neutrophils Migration and phagocytosis
Neutrophils Migration and phagocytosis
Neutrophils Phagocytosis
Neutrophil Phagocytosis
peripheral blood mononuclear cells in vitro (Bessler et al.,
1997a, 1997b). No such effect was detected in cells stimulated
by pokeweed mitogen to produce immunoglobulins. It seems
that PBR ligands are capable of suppressing spontaneous IgA
secretion but fail to affect the augmented IgA production
induced by pokeweed mitogen (Bessler et al., 1997a, 1997b).
Using mononuclear-derived cells, Klegeris et al. (2000)
found that PK 11195 inhibited the respiratory burst response,
reduced release of glutamate and interleukin-1h, and sup-
pressed secretion of products cytotoxic to neuronal cells. These
authors also suggested that PBR ligands may be useful
inhibitors of selective macrophage functions, reducing both
local and systemic inflammation. Since PK 11195 readily
enters the brain, it may be beneficial in treating central as well
as peripheral inflammatory diseases (Klegeris et al., 2000).
Indeed, it was found that PK 11195 may reduce forebrain
damage typically caused by systemic kainic acid injections
(Veenman et al., 2002).
Migration and phagocytosis of isolated human neutrophils
were increased by diazepam in a concentration-dependent
fashion (10 nM to 10 AM) (Marino et al., 2001). This
diazepam-induced migration and phagocytosis was inhibited
by PK 11195 (10 AM). In contrast, Ro5 4864 (10 nM to 10
AM) did not affect migration, but slightly enhanced phagocy-
tosis, while clonazepam, which binds to CBR and has
only little affinity for PBR, was ineffective on both parameters
at up to 10 AM (Cosentino et al., 2000; Marino et al., 2001).
The study by Marino et al. (2001) also showed that
phagocytosis induced by diazepam or Ro5 4864 was inhibited
by the Ca2+ channel blocker verapamil (10 AM), which did
not affect diazepam’s effect on migration. Competition binding
experiments performed by fluorescent staining of PBR showed
that diazepam directly interacts with PBR on human neutro-
phils (Marino et al., 2001). Both diazepam and Ro5 4864 (10
nM to 10 AM) induced a rise of intracellular free Ca2+
concentrations, which was inhibited by PK 11195 (10 AM) and
verapamil (10 AM), and prevented by extracellular Ca2+
chelation with ethylene glycol bis(2-aminoethyl ether)-
N,N,NVNV-tetraacetic acid (EGTA; 5 mmol/l) (Cosentino et
al., 2000; Marino et al., 2001). In conclusion, experimental
evidence indicates that in human neutrophils, diazepam
stimulates both migration and phagocytosis through activation
of PBR. Furthermore, diazepam-induced intracellular free Ca2+
concentration changes appear to depend on a PBR-operated,
verapamil-sensitive increase in the plasma membrane perme-
ability and subsequent extracellular Ca2+ entry. This Ca2+ entry
is suggested to contribute to diazepam-induced phagocytosis.
517
PBR ligand Effect Study
Ro5 4864, PK 11195 Down-regulation Bessler et al., 1992
Diazepam Up-regulation Marino et al., 2001
PK 11195 Down-regulation Marino et al., 2001
Ro5 4864 Up-regulation Cosentino et al., 2000; Marino et al., 2001
Midazolam Down-regulation Massoco & Palermo-Neto, 2003
The effect of diazepam on migration of neutrophils, however,
seems to occur through Ca2+-independent mechanisms (Marino
et al., 2001).
At micromolar concentrations, AHN 086 increased the
phagocytic activity of dog neutrophils (Camins et al., 1995).
Another study assayed the effects of the mixed PBR/CBR
ligand, midazolam, on equine peripheral blood neutrophils,
peritoneal macrophages, and plasma cortisol levels (Massoco
& Palermo-Neto, 2003). Midazolam induced a dose-dependent
reduction of the peripheral blood neutrophil oxidative burst and
in the capacity of both peripheral blood neutrophils to
phagocyte S. aureus. The reductions in the oxidative burst
were interpreted as a consequence of the impairment of S.
aureus-induced phagocytosis (Massoco & Palermo-Neto,
2003).
Studies on human mononuclear cells and neutrophils
suggest that PBR present in these cells indeed may play a
role in immune responses in vivo. A significant decrease
(37.5%) was found in the PBR density of mononuclear cells
from patients with osteoarthritis compared to healthy controls
(Bazzichi et al., 2003). An up-regulation of PBR density and
affinity in neutrophils of psoriasic arthritis patients compared to
healthy controls was observed (Giusti et al., 2004). In patients
affected by osteoarthritis and rheumatoid arthritis, no such
changes in PBR characteristics were detected (Giusti et al.,
2004). These studies suggest differential responses from PBR
of different types of immunoresponsive cells to various human
inflammatory diseases.
In summary, the studies presented in this last section and in
Table 6 indicate that PBRs play a role in the immune responses
of various blood cell types. In particular, PBRs may be
involved in the spontaneous secretion of interleukin-2, inter-
leukin-3, and IgA by mononuclear cells. In addition, PBR
contribution to Ca2+ cell entry may affect phagocytosis by
neutrophils. Furthermore, a role for PBR in the migration of
neutrophils has been suggested. Finally, PBRs may affect
proliferation of lymphocytes and mononuclear cells. In
conclusion, it is likely that PBRs are involved in various
aspects of blood cell immune responses.
17. Summary and conclusions
PBRs appear to be present throughout the animal
kingdom, as well as in man. PBRs are abundant in various
organs and tissues throughout the body, including the
cardiovascular system. The PBR actually is a receptor
complex, including components such as IBP, VDAC, and
518 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
ANT (Fig. 1), which is mainly present in mitochondria. PBRs
share the VDAC and ANT in common with the MPTP. PBR
functions (Fig. 1) include regulation of steroidogenesis,
apoptosis, cell proliferation, the mitochondrial membrane
potential, the mitochondrial respiratory chain, immune
responses, voltage-dependent calcium channels, responses to
stress, microglial activation, and ischemia. Endogenous
ligands for PBR include protoporphyrin IX, DBI, TTN, and
PLA2 (Fig. 1). Classical synthetic ligands for PBR include
the isoquinoline PK 11195 and the benzodiazepine Ro5 4864.
Recently, considerable effort is being spent to develop PBR
ligands with characteristics excelling those of the classical
PBR ligands. Interesting compounds include FGIN-1-27
and SSR180575, which both possess steroidogenic properties,
but while FGIN-1-27 is pro-apoptotic, SSR180575 is
anti-apoptotic.
PBRs are abundant and inhomogeneously distributed
throughout the cardiovascular system. The components of
the cardiovascular system enriched in PBR are the platelets,
erythrocytes, lymphocytes, and mononuclear cells (Fig. 2).
Furthermore, in the walls of the cardiovascular system PBRs
are abundant in endothelial cells, striated muscle cells in the
heart, smooth muscle cells of the blood vessels, and mast
cells (Fig. 2). PBRs in the walls of the cardiovascular system
appear to take part in the responses to trauma, such as
inflammation and ischemia. The mixed PBR/CBR ligand
diazepam appears to induce cardiovascular inflammation,
while the novel PBR ligand SSR180575 (an irreversible
antagonist) appears to reduce damage correlated with ische-
mia. PBRs in blood cells are responsive to such conditions as
stress and anxiety disorders. In people exposed to acute stress,
PBR density of blood cells typically is elevated. In people
suffering from anxiety disorders, PBR density of blood cells
typically is reduced. Specific groups of patients with
neurological disorders also showed alterations in PBR levels
in blood cells. Drug treatments of patients with anxiety
disorders and neurological problems usually resulted in a
return of the PBR densities to normal levels. Nonetheless,
some types of drug treatment caused blood cell PBR levels to
deviate from normal. Since blood cell PBRs appear to
participate in several aspects of the immune response, such
as migration, phagocytosis, and interleukin-2, interleukin-3,
and IgA secretion, alterations in PBR density of the blood
cells may have consequences for the immunological condi-
tions of the patients in question.
In recent research, the involvement of PBR in inflamma-
tion, ischemia, and the immune response of particular
components of the cardiovascular system has received some
attention. Potentially, it may also be interesting to study the
involvement of PBR in repair mechanisms after injury,
angiogenesis, including angiogenesis of tumors, or formation
of blood cells. Since potent novel PBR ligands were and are
being developed, this kind or research may provide new
insights into these functions related to the cardiovascular
system. It may be envisaged that such PBR ligands may be
used for future treatments of cardiovascular diseases and
injuries, as well as to inhibit tumor angiogenesis.
Acknowledgments
The Center for Absorption in Science, Ministry of Immi-
grant Absorption, State of Israel, is acknowledged for its
support to L.V.
We thank Dr. Svetlana Leschiner for her aid with the
preparation of the manuscript. We also thank Mrs. Lena Moine
for her advice to Fig. 1.
References
Abbott, N. J. (2000). Inflammatory mediators and modulation of blood-brain
barrier permeability. Cell Mol Neurobiol 20, 131 – 147.
Alexander, B. E., Roller, E., & Klotz, U. (1992). Characterization of
peripheral-type benzodiazepine binding sites on human lymphocytes and
lymphoma cell lines and their role in cell growth. Biochem Pharmacol 44,
269 – 274.
Alho, H., Fremeau Jr, R. T., Tiedge, H., Wilcox, J., Bovolin, P., Brosius, J., et al.
(1988). Diazepam binding inhibitor gene expression: location in brain and
peripheral tissues of rat. Proc Natl Acad Sci U S A 85, 7018 – 7022.
Anholt, R. R., De Souza, E. B., Oster-Granite, M. L., & Snyder, S. H.
(1985). Peripheral-type benzodiazepine receptors: autoradiographic local-
ization in whole-body sections of neonatal rats. J Pharmacol Exp Ther
233, 517 – 526.
Anholt, R. R. H., Pedersen, P. L., De Souza, E. B., & Snyder, S. H. (1986). The
peripheral-type benzodiazepine receptor: localization to the mitochondrial
outer membrane. J Biol Chem 261, 576 – 583.
Antkiewicz-Michaluk, L., Guidotti, A., & Krueger, K. E. (1988).
Molecular characterization and mitochondrial density of a recognition
site for peripheral-type benzodiazepine ligands. Mol Pharmacol 34,
272 – 278.
Avital, A., Richter-Levin, G., Leschiner, S., Spanier, I., Veenman, L., Weizman,
A., et al. (2001). Acute and repeated swim stress effects on peripheral
benzodiazepine receptors in the rat hippocampus, adrenal, and kidney.
Neuropsychopharmacology 25, 669 – 678.
Awad, M., & Gavish, M. (1987). Binding of [3H]Ro 5-4864 and [3H]PK 11195
to cerebral cortex and peripheral tissues of various species: species
differences and heterogeneity in peripheral benzodiazepine binding sites.
J Neurochem 49, 1407 – 1414.
Awad, M., & Gavish, M. (1991). Peripheral-type benzodiazepine receptors in
human cerebral cortex, kidney, and colon. Life Sci 49, 1155 – 1161.
Ball, J. A., Ghatei, M. A., Sekiya, K., Krausz, T., & Bloom, S. R. (1989).
Diazepam binding inhibitor-like immunoreactivity(51 – 70): distribution in
human brain, spinal cord and peripheral tissues. Brain Res 479, 300 – 305.
Bar-Ami, S., Fares, F., & Gavish, M. (1989). Effect of hypophysectomy
and hormone treatment on the induction of peripheral-type benzodiaz-
epine binding sites in female rat genital axis. Horm Metab Res 21,
106 – 107.
Basile, A. S., Weissman, B. A., & Skolnick, P. (1987). Maximal electroshock
increases the density of [3H]Ro 5-4864 binding to mouse cerebral cortex.
Brain Res Bull 19, 1 – 7.
Bazzichi, L., Betti, L., Giannaccini, G., Rossi, A., & Lucacchini, A. (2003).
Peripheral-type benzodiazepine receptors in human mononuclear cells of
patients affected by osteoarthritis, rheumatoid arthritis or psoriasic arthritis.
Clin Biochem 36, 57 – 60.
Belloli, S., Moresco, R. M., Matarrese, M., Biella, G., Sanvito, F., Simonelli, P.,
et al. (2004). Evaluation of three quinoline-carboxamide derivatives as
potential radioligands for the in vivo pet imaging of neurodegeneration.
Neurochem Int 44, 433 – 440.
Belzacq, A. S., & Brenner, C. (2003). The adenine nucleotide translocator: a
new potential chemotherapeutic target. Curr Drug Targets 4, 517 – 524.
Benavides, J., Guilloux, F., Rufat, P., Uzan, A., Renault, C., Dubroeucq, M. C.,
et al. (1984). In vivo labelling in several rat tissues of Fperipheral type_
benzodiazepine binding sites. Eur J Pharmacol 99, 1 – 7.
Benavides, J., Quarteronet, D., Plouin, P. F., Imbault, F., Phan, T., Uzan, A.,
et al. (1984). Characterization of peripheral type benzodiazepine binding
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
sites in human and rat platelets by using [3H]PK 11195. Studies in
hypertensive patients. Biochem Pharmacol 33, 2467 – 2472.
Berkovich, A., Ferrarese, C., Cavaletti, G., Alho, H., Marzorati, C., Bianchi, G.,
et al. (1993). Topology of two DBI receptors in human lymphocytes. Life
Sci 52(15), 1265 – 1277.
Bessler, H., Weizman, R., Gavish, M., Notti, I., & Djaldetti, M. (1992).
Immunomodulatory effect of peripheral benzodiazepine receptor ligands on
human mononuclear cells. J Neuroimmunol 38, 19 – 25.
Bessler, H., Caspi, B., Gavish, M., Rehavi, M., & Weizman, A. (1997a).
Significant inhibition of spontaneous IgA secretion by selective peripheral-
type benzodiazepine receptor ligands. Clin Neuropharmacol 20, 215 – 223.
Bessler, H., Caspi, B., Gavish, M., Rehavi, M., & Weizman, A. (1997b).
Significant inhibition of spontaneous IgA secretion by selective peripheral-
type benzodiazepine receptor ligands. Clin Neuropharmacol 20, 215 – 223.
Betti, L., Giannaccini, G., Nigro, M., Dianda, S., Gremigni, V., & Lucacchini,
A. (2003). Studies of peripheral benzodiazepine receptors in mussels:
comparison between a polluted and a nonpolluted site. Ecotoxicol Environ
Saf 54, 36 – 42.
Bidder, M., Ratzoni, G., Weizman, A., Blumensohn, R., Norymberg, M.,
Tyano, S., et al. (1990). Platelet benzodiazepine binding in Alzheimer’s
disease. Biol Psychiatry 28, 641 – 643.
Blahos Jr., J., Whalin, M. E., & Krueger, K. E. (1995). Identification and
purification of a 10-kilodalton protein associated with mitochondrial
benzodiazepine receptors. J Biol Chem 270, 20285 – 20291.
Blandini, F., Cosentino, M., Mangiagalli, A., Marino, F., Samuele, A., Rasini,
E., et al. (2004). Modifications of apoptosis-related protein levels in
lymphocytes of patients with Parkinson’s disease. The effect of dopami-
nergic treatment. J Neural Transm 111, 1017 – 1030.
Bolger, G. T., Weissman, B. A., Lueddens, H., Barrett, J. E., Witkin, J., Paul, S.
M., et al. (1986). Dihydropyridine calcium channel antagonists binding in
non-mammalian vertebrates: characterization and relationship to Fperipheral-
type_ binding sites for benzodiazepines. Brain Res 368, 351 – 356.
Bolger, G. T., Newman, A. H., Rice, K. C., Lueddens, H. W., Basile, A. S., &
Skolnick, P. (1989). Characterization of the effects of AHN 086, an
irreversible ligand of ‘‘peripheral’’ benzodiazepine receptors, on contraction
in guinea-pig atrial and ileal longitudinal smooth muscle. Can J Physiol
Pharm 67, 126 – 134.
Bolger, G. T., Abraham, S., Oz, N., & Weissman, B. A. (1990). Interactions
between peripheral-type benzodiazepine receptor ligands and an activator of
voltage-operated calcium channels. Can J Physiol Pharm 68, 40 – 45.
Bongioanni, P., Donato, M., Castagna, M., & Gemignani, F. (1996). Platelet
phenolsulphotransferase activity, monoamine oxidase activity and periph-
eral-type benzodiazepine binding in demented patients. J Neural Transm
103, 491 – 501.
Bongioanni, P., Castagna, M., Mondino, C., Boccardi, B., & Borgna, M.
(1997). Platelet and lymphocyte benzodiazepine binding in patients with
Alzheimer’s disease. Exp Neurol 146, 560 – 566.
Bono, F., Lamarche, I., Prabonnaud, V., Le Fur, G., & Herbert, J. M. (1999).
Peripheral benzodiazepine receptor agonists exhibit potent antiapoptotic
activities. Biochem Biophys Res Commun 265, 457 – 461.
Bono, F., Herbert, J. M., & Parini, A. (2004). Involvement of peripheral
benzodiazepine receptor in the oxidative stress, death-signaling pathways,
and renal injury induced by ischemia-reperfusion. J Am Soc Nephrol 15,
2152 – 2160.
Bonuccelli, U., Nuti, A., Del Dotto, P., Piccini, P., Martini, C., Giannaccini, G.,
et al. (1991). Platelet peripheral benzodiazepine receptors are decreased in
Parkinson’s disease. Life Sci 48, 1185 – 1190.
Bovolin, P., Schlichting, J., Miyata, M., Ferrarese, C., Guidotti, A., & Alho, H.
(1990). Distribution and characterization of diazepam binding inhibitor
(DBI) in peripheral tissues of rat. Regul Pept 29, 267 – 281.
Braestrup, C., & Squires, R. F. (1977). Specific benzodiazepine receptors in rat
brain characterized by high-affinity [3H]diazepam binding. Proc Natl Acad
Sci U S A 74, 3805 – 3809.
Bribes, E., Casellas, P., Vidal, H., Dussossoy, D., & Casellas, D. (2002).
Peripheral benzodiazepine receptor mapping in rat kidney. Effects of
angiotensin II-induced hypertension. J Am Soc Nephrol 13, 1 – 9.
Brown, R. C., Degenhardt, B., Kotoula, M., & Papadopoulous, V. (2000).
Location-dependent role of the human glioma cell peripheral-type benzo-
519
diazepine receptor in proliferation and steroid biosynthesis. Cancer Lett
156, 125 – 132.
Cahard, D., Canat, X., Carayon, P., Roque, C., Casellas, P., & Le Fur, G.
(1994). Subcellular localization of peripheral benzodiazepine receptors on
human leukocytes [comment in Lab Invest, 1994;70:1 – 5]. Lab Invest 70,
23 – 28.
Camins, A., Escubedo, E., Camarasa, J., Carretero, J., Newman, A. H., &
Kammuller, M. E. (1995). Dog neutrophil stimulation and cell surface
expression of heat shock proteins induced by AHN 086. J Pharmacol Exp
Ther 274, 242 – 248.
Campagna, F., Palluotto, F., Carotti, A., & Maciocco, E. (2004). Synthesis,
central and peripheral benzodiazepine receptor affinity of pyrazole and
pyrazole-containing polycyclic derivatives. Farmaco 59, 849 – 856.
Campiani, G., Fiorini, I., De Filippis, M. P., Ciani, S. M., Garofalo, A., Nacci,
V., et al. (1996a). Cardiovascular characterization of pyrrolo[2,1-
d][1,5]benzothiazepine derivatives binding selectively to the peripheral-
type benzodiazepine receptor (PBR): from dual PBR affinity and calcium
antagonist activity to novel and selective calcium entry blockers. J Med
Chem 39, 2922 – 2938.
Campiani, G., Nacci, V., Fiorini, I., De Filippis, M. P., Garofalo, A., Ciani, S.
M., et al. (1996b). Synthesis, biological activity, and SARs of pyrroloben-
zoxazepine derivatives, a new class of specific ‘‘peripheral-type’’ benzodi-
azepine receptor ligands. J Med Chem 39, 3435 – 3450.
Campiani, G., Ramunno, A., Fiorini, I., Nacci, V., Morelli, E., Novellino, E.,
et al. (2002). Synthesis of new molecular probes for investigation of steroid
biosynthesis induced by selective interaction with peripheral type benzo-
diazepine receptors (PBR). J Med Chem 45, 4276 – 4281.
Canat, X., Carayon, P., Bouaboula, M., Cahard, D., Shire, D., Roque, C., et al.
(1993). Distribution profile and properties of peripheral-type benzodiaze-
pine receptors on human hemopoietic cells. Life Sci 52, 107 – 118.
Cappelli, A., Anzini, M., Vomero, S., De Benedetti, P. G., Menziani, M. C.,
Giorgi, G., et al. (1997). Mapping the peripheral benzodiazepine receptor
binding site by conformationally restrained derivatives of 1-(2-chlorophe-
nyl)-N-methyl-N-(1-methylpropyl)-3-isoquinolinecarboxamide (PK11195).
J Med Chem 40, 2910 – 2921.
Cappelli, A., Pericot Mohr, G., Gallelli, A., Giuliani, G., Anzini, M., Vomero,
S., et al. (2003). Structure-activity relationships in carboxamide derivatives
based on the targeted delivery of radionuclides and boron atoms by means of
peripheral benzodiazepine receptor ligands. J Med Chem 46, 3568 – 3571.
Carmel, I., Fares, F. A., Leschiner, S., Scherubl, H., Weisinger, G., & Gavish,
M. (1999). Peripheral-type benzodiazepine receptors in the regulation of
proliferation of MCF-7 human breast carcinoma cell line. Biochem
Pharmacol 58, 273 – 278.
Chelli, B., Falleni, A., Salvetti, F., Gremigni, V., Lucacchini, A., & Martini, C.
(2001). Peripheral-type benzodiazepine receptor ligands: mitochondrial
permeability transition induction in rat cardiac tissue. Biochem Pharmacol
61, 695 – 705.
Chelli, B., Lena, A., Vanacore, R., Pozzo, E. D., Costa, B., Rossi, L., et al.
(2004). Peripheral benzodiazepine receptor ligands: mitochondrial trans-
membrane potential depolarization and apoptosis induction in rat C6 glioma
cells. Biochem Pharmacol 68, 125 – 134.
Chen, Y., Zheng, X., Dobhal, M. P., Gryshuk, A., Morgan, J., Dougherty,
T. J., et al. (2005). Methyl pyropheophorbide-a analogues: potential
fluorescent probes for the peripheral-type benzodiazepine receptor.
Effect of central metal in photosensitizing efficacy. J Med Chem 48,
3692 – 3695.
Chiu, S., Singh, A. N., Chiu, P., & Mishra, R. K. (2001). Inverse agonist
binding of peripheral benzodiazepine receptors in anxiety disorder. Eur
Arch Psychiatry Clin Neurosci 251, 136 – 140.
Conway, E. L., Gundlach, A. L., & Craven, J. A. (1998). Temporal changes in
glial fibrillary acidic protein messenger RNA and [3H]PK11195 binding in
relation to imidazoline-I2-receptor and alpha 2-adrenoceptor binding in the
hippocampus following transient global forebrain ischaemia in the rat.
Neuroscience 82, 805 – 817.
Cosentino, M., Marino, F., Cattaneo, S., Di Grazia, L., Francioli, C., Fietta, A.
M., et al. (2000). Diazepam-binding inhibitor-derived peptides induce
intracellular calcium changes and modulate human neutrophil function. J
Leukoc Biol 67, 637 – 643.
520 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Cox, D. A., Ellinor, P. T., Kirley, T. L., & Matlib, M. A. (1991). Identification
of a 17-kDa protein associated with the peripheral-type benzodiazepine
receptor in vascular and other smooth muscle types. J Pharmacol Exp Ther
258, 702 – 709.
Cremer, J. E., Hume, S. P., Cullen, B. M., Myers, R., Manjil, L. G., Turton, D.
R., et al. (1992). The distribution of radioactivity in brains of rats given [N-
methyl-11C]PK 11195 in vivo after induction of a cortical ischaemic lesion.
Int J Radiat Appl Instrum B 19, 159 – 166.
Culty, M., Li, H., Boujrad, N., Amri, H., Vidic, B., Bernassau, J. M., et al.
(1999). In vitro studies on the role of the peripheral-type benzodiaz-
epine receptor in steroidogenesis. J Steroid Biochem Mol Biol 69,
123 – 130.
Daly, R. C., Schmidt, P. J., Davis, C. L., Danaceau, M. A., & Rubinow, D. R.
(2001). Effects of gonadal steroids on peripheral benzodiazepine receptor
density in women with PMS and controls. Psychoneuroendocrinology 26,
539 – 549.
Dar, D. E., Weizman, A., Karp, L., Grinshpoon, A., Bidder, M., Kotler, M., et al.
(1991). Platelet peripheral benzodiazepine receptors in repeated stress. Life
Sci 48, 341 – 346.
Daval, J. L., Deckert, J., Nakajima, T., Morgan, P. F., & Marangos, P. J. (1988).
Regional ontogenetic profile of central and peripheral benzodiazepine
receptors in guinea pig brain. Neurosci Lett 92, 82 – 85.
Davies, L. P., & Huston, V. (1981). Peripheral benzodiazepine binding sites
in heart and their interaction with dipyridamole. Eur J Pharmacol 73,
209 – 211.
Demerle-Pallardy, C., Duverger, D., Spinnewyn, B., Pirotzky, E., & Braquet, P.
(1991). Peripheral type benzodiazepine binding sites following transient
forebrain ischemia in the rat: effect of neuroprotective drugs. Brain Res
565, 312 – 320.
Dolder, M., Wendt, S., & Wallimann, T. (2001). Mitochondrial creatine kinase
in contact sites: interaction with porin and adenine nucleotide translocase,
role in permeability transition and sensitivity to oxidative damage. Biol
Signals Recept 10, 93 – 111.
Drobinski, G., Lechat, P., Eugene, L., & Grosgogeat, Y. (1989). Absence of
an anti-ischemic effect of the peripheral benzodiazepine receptor
antagonist PK 11195. Study by atrial stimulation in man. Therapies 44,
263 – 267.
Droogleever Fortuyn, H. A., van Broekhoven, F., Span, P. N., Backstrom,
T., Zitman, F. G., & Verkes, R. J. (2004). Effects of PhD examination
stress on allopregnanolone and cortisol plasma levels and peripheral
benzodiazepine receptor density. Psychoneuroendocrinology 29,
1341 – 1344.
Drugan, R. C., Basile, A. S., Crawley, J. N., Paul, S. M., & Skolnick, P. (1988).
Characterization of stress-induced alterations in [3H] Ro5-4864 binding to
peripheral benzodiazepine receptors in rat heart and kidney. Pharmacol
Biochem Behav 30, 1015 – 1020.
Dumont, F., De Vos, F., Versijpt, J., Jansen, H. M., Korf, J., Dierckx, R. A., et al.
(1999). In vivo evaluation in mice and metabolism in blood of human
volunteers of [123I]iodo-PK11195: a possible single-photon emission
tomography tracer for visualization of inflammation. Eur J Nucl Med 26,
194 – 200.
Edoute, Y., Giris, J., Ben-Haim, S. A., Lochner, A., Weizman, A., Hayam,
G., et al. (1993). Ro 5-4864 and PK 11195, but not diazepam, depress
cardiac function in an isolated working rat heart model. Pharmacology
46, 224 – 230.
Eshleman, A. J., & Murray, T. F. (1989). Differential binding properties of the
peripheral-type benzodiazepine ligands [3H]PK 11195 and [3H]Ro 5-4864
in trout and mouse brain membranes. J Neurochem 53, 494 – 502.
Fares, F., Weizman, A., Zlotogorski, D., & Gavish, M. (1987a). Ontogenic
development of peripheral benzodiazepine binding sites in rat brain, heart
and lung. Brain Res 408, 381 – 384.
Fares, F., Bar-Ami, S., Brandes, J. M., & Gavish, M. (1987b).
Gonadotropin- and estrogen-induced increase of peripheral-type benzo-
diazepine binding sites in the hypophyseal-genital axis of rats. Eur J
Pharmacol 133, 97 – 102.
Fares, F., Bar-Ami, S., Brandes, J. M., & Gavish, M. (1988). Changes in the
density of peripheral benzodiazepine binding sites in genital organs of the
female rat during the oestrous cycle. J Reprod Fertil 83, 619 – 625.
Fares, F., Bar-Ami, S., Haj-Yehia, Y., & Gavish, M. (1989). Hormonal
regulation of peripheral benzodiazepine binding sites in female rat adrenal
gland and kidney. J Recept Res 9, 143 – 157.
Faure, J. P., Hauet, T., Han, Z., Goujon, J. M., Petit, I., Mauco, G., et al. (2002).
Polyethylene glycol reduces early and long-term cold ischemia-reperfusion
and renal medulla injury. J Pharmacol Exp Ther 302, 861 – 870.
Faure, J. P., Baumert, H., Han, Z., Goujon, J. M., Favreau, F., Dutheil, D.,
et al. (2003). Evidence for a protective role of trimetazidine during cold
ischemia: targeting inflammation and nephron mass. Biochem Pharmacol
66, 2241 – 2250.
Ferrarese, C., Perego, M., Marzorati, C., Bianchi, G., Frigo, M., Pecora, N.,
et al. (1996). Modifications of diazepam binding inhibitor and peripheral
benzodiazepine receptors in the lymphocytes of epileptic patients. J Neurol
Sci 17, 141 – 145.
Ferrarese, C., Tortorella, R., Bogliun, G., Beghi, E., Cogliati, T., Zoia, C., et al.
(1997). Decreased density of lymphocyte benzodiazepine receptors in drug-
resistant epileptic patients. Epilepsy Res 27, 181 – 185.
Ferrero, P., Rocca, P., Benna, P., De Leo, C., Montalenti, E., Ravizza, L., et al.
(1992). An analysis of peripheral type benzodiazepine receptors on blood
mononuclear cells during high dose steroid treatment of multiple sclerosis.
Ital J Neurol Sci 13, 685 – 691.
Ferzaz, B., Brault, E., Bourliaud, G., Robert, J. P., Poughon, G., Claustre, Y.,
et al. (2002). SSR180575 (7-chloro-N,N,5-trimethyl-4-oxo-3-phenyl-3,5-
dihydro-4H-pyridazino[4,5-b]indole-1-acetamide), a peripheral benzodiaz-
epine receptor ligand, promotes neuronal survival and repair. J Pharmacol
Exp Ther 301, 1067 – 1078.
File, S. E., & Pellow, S. (1983). RO5-4864, a ligand for benzodiazepine
micromolar and peripheral binding sites: antagonism and enhancement of
behavioural effects. Psychopharmacology (Berl) 80, 166 – 170.
French, J. F., & Matlib, M. A. (1988). Identification of a high-affinity
peripheral-type benzodiazepine binding site in rat aortic smooth muscle
membranes. J Pharmacol Exp Ther 247, 23 – 28.
French, J. F., Rapoport, R. M., & Matlib, M. A. (1989). Possible mechanism of
benzodiazepine-induced relaxation of vascular smooth muscle. J Cardio-
vasc Pharmacol 14, 405 – 411.
Galdiero, F., Bentivoglio, C., Nuzzo, I., Ianniello, R., Capasso, C., Mattera, S.,
et al. (1995). Effects of benzodiazepines on immunodeficiency and
resistance in mice. Life Sci 57, 2413 – 2423.
Galiegue, S., Jbilo, O., Combes, T., Bribes, E., Carayon, P., Le Fur, G., et al.
(1999). Cloning and characterization of PRAX-1. A new protein that
specifically interacts with the peripheral benzodiazepine receptor. J Biol
Chem 274, 2938 – 2952.
Galiegue, S., Tinel, N., & Casellas, P. (2003). The peripheral benzodiaz-
epine receptor: a promising therapeutic drug target. Curr Med Chem 10,
1563 – 1572.
Garnier, M., Dimchev, A. B., Boujrad, N., Price, J. M., Musto, N. A., &
Papadopoulos, V. (1994). In vitro reconstitution of a functional peripheral-
type benzodiazepine receptor from mouse Leydig tumor cells. Mol
Pharmacol 45, 201 – 211.
Gavish, M., Weizman, A., Karp, L., Tyano, S., & Tanne, Z. (1986). Decreased
peripheral benzodiazepine binding sites in platelets of neuroleptic-treated
schizophrenics. Eur J Pharmacol 121, 275 – 279.
Gavish, M., Katz, Y., Bar-Ami, S., & Weizman, R. (1992). Biochemical,
physiological, and pathological aspects of the peripheral benzodiazepine
receptor. J Neurochem 58, 1589 – 1601.
Gavish, M., Laor, N., Bidder, M., Fisher, D., Fonia, O., Muller, U.,
et al. (1996). Altered platelet peripheral-type benzodiazepine receptor
in posttraumatic stress disorder. Neuropsychopharmacology 14,
181 – 186.
Gavish, M., Bachman, I., Shoukrun, R., Katz, Y., Veenman, L., Weisinger, G.,
et al. (1999). Enigma of the peripheral benzodiazepine receptor. Pharmacol
Rev 51, 629 – 650.
Gazouli, M., Han, Z., & Papadopoulos, V. (2002). Identification of a peptide
antagonist to the peripheral-type benzodiazepine receptor that inhibits
hormone-stimulated leydig cell steroid formation. J Pharmacol Exp Ther
303, 627 – 632.
Gehlert, D. R., Yamamura, H. I., & Wamsley, J. K. (1985). Autoradiographic
localization of ‘‘peripheral-type’’ benzodiazepine binding sites in the rat
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
brain, heart and kidney. Naunyn-Schmiedeberg’s Arch Pharmacol 328,
454 – 460.
Giannaccini, G., Martini, C., Lucacchini, A., Strettoi, E., & Piccolino, M.
(1993). The peripheral-type benzodiazepine receptor ligands [3H]Ro 5-4864
and [3H]PK 11195 bind to the retina of rabbit, but not of turtle. J Neurochem
61, 1263 – 1269.
Gildersleeve, D. L., Van Dort, M. E., Johnson, J. W., Sherman, P. S., &
Wieland, D. M. (1996). Synthesis and evaluation of [123I]-iodo-PK11195
for mapping peripheral-type benzodiazepine receptors (omega 3) in heart.
Nucl Med Biol 23, 23 – 28.
Gimeno, M., Pallas, M., Newman, A. H., Camarasa, J., & Escubedo, E. (1994).
The role of cyclic nucleotides in the action of peripheral-type benzodiaz-
epine receptor ligands in rat aorta. Gen Pharmacol 25, 1553 – 1561.
Giusti, L., Betti, L., Giannaccini, G., Mascia, G., Bazzichi, L., & Lucacchini,
A. (2004). [3H]PK11195 binding sites in human neutrophils: effect of
fMLP stimulation and modulation in rheumatic diseases. Clin Biochem
37, 61 – 66.
Godlewski, M. M., Gajkowska, B., Lamparska-Przybysz, M., & Motyl, T.
(2002). Colocalization of BAX with BID and VDAC-1 in nimesulide-
induced apoptosis of human colon adenocarcinoma COLO 205 cells.
Anticancer Drugs 13, 1017 – 1029.
Golani, I., Weizman, A., Leschiner, S., Spanier, I., Eckstein, N., Limor, R.,
et al. (2001). Hormonal regulation of peripheral benzodiazepine receptor
binding properties is mediated by subunit interaction. Biochemistry 40,
10213 – 10222.
Graham, B. H., & Craigen, W. J. (2005). Mitochondrial voltage-dependent
anion channel gene family in Drosophila melanogaster: complex patterns
of evolution, genomic organization, and developmental expression. Mol
Genet Metab 85, 308 – 317.
Graham, C. W., Pagano, R. R., & Katz, R. L. (1977). Thrombophlebitis after
intravenous diazepam-can it be prevented? Anesth Analg 56, 409 – 413.
Hall, E. D., Andrus, P. K., Fleck, T. J., Oostveen, J. A., Carter, D. B., &
Jacobsen, E. J. (1997). Neuroprotective properties of the benzodiazepine
receptor, partial agonist PNU-101017 in the gerbil forebrain ischemia
model. J Cereb Blood Flow Metab 17, 875 – 883.
Hardwick, M., Fertikh, D., Culty, M., Li, H., Vidic, B., & Papadopoulos, V.
(1999). Peripheral-type benzodiazepine receptor (PBR) in human breast
cancer: correlation of breast cancer cell aggressive phenotype with PBR
expression, nuclear localization, and PBR-mediated cell proliferation and
nuclear transport of cholesterol. Cancer Res 59, 831 – 842.
Hashimoto, K., Inoue, O., Suzuki, K., Yamasaki, T., & Kojima, M. (1989).
Synthesis and evaluation of 11C-PK 11195 for in vivo study of peripheral-
type benzodiazepine receptors using positron emission tomography. Ann
Nucl Med 3, 63 – 71.
Hauet, T., Han, Z., Wang, Y., Hameury, F., Jayle, C., Gibelin, H., et al. (2002).
Modulation of peripheral-type benzodiazepine receptor levels in a reperfu-
sion injury pig kidney-graft model. Transplantation 74, 1507 – 1515.
Hirsch, J. D., Beyer, CF., Malkowitz, L., Beer, B., & Blume, A. J. (1989).
Mitochondrial benzodiazepine receptors mediate inhibition of mitochon-
drial respiratory control. Mol Pharmacol 35, 157 – 163.
Hollingsworth, E. B., McNeal, E. T., Burton, J. L., Williams, R. J., Daly, J. W., &
Creveling, C. R. (1985). Biochemical characterization of a filtered synapto-
neurosome preparation from guinea pig cerebral cortex: cyclic adenosine 3V:5V-
monophosphate-generating systems, receptors, and enzymes. J Neurosci 5,
2240 – 2253.
Ikezaki, K., Black, K. L., Santori, E. M., Smith, M. L., Becker, D. P., Payne, B.
A., et al. (1990). Three-dimensional comparison of peripheral benzodiaz-
epine binding and histological findings in rat brain tumor. Neurosurgery 27,
78 – 82.
Itzhak, Y., Roig-Cantisano, A., & Norenberg, M. D. (1995). Ontogeny of
peripheral-type benzodiazepine receptors in cultured astrocytes and brain
from rat. Brain Res Dev Brain Res 84, 62 – 66.
Johnson, M. R., Marazziti, D., Brawman-Mintzer, O., Emmanuel, N. P., Ware,
M. R., Morton, W. A., et al. (1998). Abnormal peripheral benzodiazepine
receptor density associated with generalized social phobia. Biol Psychiatry
43, 306 – 309.
Jorda, E. G., Jimenez, A., Verdaguer, E., Canudas, A. M., Folch, J., Sureda, F.
X., et al. (2005). Evidence in favour of a role for peripheral-type
521
benzodiazepine receptor ligands in amplification of neuronal apoptosis.
Apoptosis 10, 91 – 104.
Karp, L., Weizman, A., Tyano, S., & Gavish, M. (1989). Examination stress,
platelet peripheral benzodiazepine binding sites, and plasma hormone
levels. Life Sci 44, 1077 – 1082.
Karp, L., Weizman, A., Fliman, M., Tyano, S., & Gavish, M. (1991). Peripheral
benzodiazepine receptors on platelets in chronic and detoxified alcoholics.
Neuropharmacology 30, 665 – 669.
Kelly-Hershkovitz, E., Weizman, R., Spanier, I., Leschiner, S., Lahav, M.,
Weisinger, G., et al. (1998). Effects of peripheral-type benzodiazepine
receptor antisense knockout on MA-10 Leydig cell proliferation and
steroidogenesis. J Biol Chem 273, 5478 – 5483.
Klegeris, A., McGeer, E. G., & McGeer, P. L. (2000). Inhibitory action of 1-(2-
chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinolinecarboxamide
(PK 11195) on some mononuclear phagocyte functions. Biochem Pharma-
col 59, 1305 – 1314.
Kletsas, D., Li, W., Han, Z., & Papadopoulos, V. (2004). Peripheral-type
benzodiazepine receptor (PBR) and PBR drug ligands in fibroblast and
fibrosarcoma cell proliferation: role of ERK, c-Jun and ligand-activated
PBR-independent pathways. Biochem Pharmacol 67, 1927 – 1932.
Koiso, Y., Nakajima, O., Matsumura, D., Fujimoto, Y., & Hashimoto, Y.
(2000). Chemical control of cell differentiation of human myeloleukemia
K562 cell line. Yakushigaku Zasshi 120, 104 – 112.
Kooistra, T., Toet, K., Kluft, C., VonVoigtlander, P. F., Ennis, M. D., Aiken,
J. W., et al. (1993). Triazolobenzodiazepines: a new class of stimulators
of tissue-type plasminogen activator synthesis in human endothelial cells.
Biochem Pharmacol 46, 61 – 67.
Kragie, L., & Smiehorowski, R. (1994). Altered peripheral benzodiazepine
receptor binding in cardiac and liver tissues from thyroidectomized rats.
Life Sci 55, 1911 – 1918.
Krueger, K. E. (1995). Molecular and functional properties of mitochondrial
benzodiazepine receptors. Biochim Biophys Acta 1241, 453 – 470.
Kunduzova, O. R., Escourrou, G., De La Farge, F., Salvayre, R., Seguelas,
M. H., Leducq, N., et al. (2004). Involvement of peripheral benzodiaz-
epine receptor in the oxidative stress, death-signaling pathways, and
renal injury induced by ischemia-reperfusion. J Am Soc Nephrol 15,
2152 – 2160.
Lacapere, J. J., & Papadopoulos, V. (2003). Peripheral-type benzodiazepine
receptor: structure and function of a cholesterol-binding protein in steroid
and bile acid biosynthesis. Steroids 68, 569 – 585.
Landau, M., Weizman, A., Zoref-Shani, E., Beery, E., Wasseman, L., Landau,
O., et al. (1998). Antiproliferative and differentiating effects of benzodiaz-
epine receptor ligands on B16 melanoma cells. Biochem Pharmacol 56,
1029 – 1034.
Lazzarini, R., Malucelli, B. E., & Palermo-Neto, J. (2001). Reduction of
acute inflammation in rats by diazepam: role of peripheral benzodiazepine
receptors and corticosterone. Immunopharmacol Immunotoxicol 23,
253 – 265.
Leducq, N., Bono, F., Sulpice, T., Vin, V., Janiak, P., Le Fur, G., et al. (2003).
Role of peripheral benzodiazepine receptors in mitochondrial, cellular and
cardiac damage induced by oxidative stress and ischemia-reperfusion. J
Pharmacol Exp Ther 306, 828 – 837.
Le Fur, G., Perrier, M. L., Vaucher, N., Imbault, F., Flamier, A., Benavides, J.,
et al. (1983a). Peripheral benzodiazepine binding sites: effect of PK 11195,
1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinolinecarboxa-
mide: I. In vitro studies. Life Sci 32, 1839 – 1847.
Le Fur, G., Guilloux, F., Rufat, P., Benavides, J., Uzan, A., Renault, C., et al.
(1983b). Peripheral benzodiazepine binding sites: effect of PK 11195, 1-(2-
chlorophenyl)-N-methyl-(1-methylpropyl)-3 isoquinolinecarboxamide: II.
In vivo studies. Life Sci 32, 1849 – 1856.
Le Fur, G., Vaucher, N., Perrier, M. L., Flamier, A., Benavides, J., Renault, C.,
et al. (1983c). Differentiation between two ligands for peripheral benzodi-
azepine binding sites, [3H]RO5-4864 and [3H]PK 11195, by thermody-
namic studies. Life Sci 33, 449 – 457.
Lesouhaitier, O., Feuilloley, M., Lihrmann, I., Ugo, I., Fasolo, A., Tonon, M.
C., et al. (1996). Localization of diazepam-binding inhibitor-related
peptides and peripheral type benzodiazepine receptors in the frog adrenal
gland. Cell Tissue Res 283, 403 – 412.
522 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Levin, E., Premkumar, A., Veenman, L., Kugler, W., Leschiner, S., Spanier, I.,
et al. (2005). The peripheral-type benzodiazepine receptor and tumorige-
nicity: isoquinoline binding protein (IBP) antisense knockdown in the C6
glioma cell line. Biochemistry 44, 9924 – 9935.
Lheureux, P., Vranckx, M., Leduc, D., & Askenasi, R. (1990). Effect of
PK11195 on cardiovascular toxicity due to verapamil: an experimental
study in the dog. J Toxicol Clin Exp 10, 449 – 460.
Li, H., Degenhardt, B., Tobin, D., Yao, Z. X., Tasken, K., & Papadopoulos, V.
(2001). Identification, localization, and function in steroidogenesis of
PAP7: a peripheral-type benzodiazepine receptor- and PKA (RIalpha)-
associated protein. Mol Endocrinol 15, 2211 – 2228.
Liu, J., Li, H., & Papadopoulos, V. (2003). PAP7, a PBR/PKA-RIalpha-
associated protein: a new element in the relay of the hormonal induction of
steroidogenesis. J Steroid Biochem Mol Biol 85, 275 – 283.
Lueddens, H. W., Newman, A. H., Rice, K. C., & Skolnick, P. (1986). AHN
086: an irreversible ligand of ‘‘peripheral’’ benzodiazepine receptors. Mol
Pharmacol 29, 540 – 545.
Maaser, K., Hopfner, M., Jansen, A., Weisinger, G., Gavish, M., Kozikowski,
A. P., et al. (2001). Specific ligands of the peripheral benzodiazepine
receptor induce apoptosis and cell cycle arrest in human colorectal cancer
cells. Br J Cancer 85, 1771 – 1780.
Maaser, K., Sutter, A. P., & Scherubl, H. (2005). Mechanisms of mitochondrial
apoptosis induced by peripheral benzodiazepine receptor ligands in human
colorectal cancer cells. Biochem Biophys Res Commun 332, 646 – 652.
Maeda, S., Miyawaki, T., Nakanishi, T., Takigawa, M., & Shimada, M. (1998).
Peripheral type benzodiazepine receptor in T lymphocyte rich preparation.
Life Sci 63, 1423 – 1430.
Maeda, J., Suhara, T., Zhang, M. R., Okauchi, T., Yasuno, F., Ikoma, Y., et al.
(2004). Novel peripheral benzodiazepine receptor ligand [11C]DAA1106
for PET: an imaging tool for glial cells in the brain. Synapse 52, 283 – 291.
Mak, J. C., & Barnes, P. J. (1990). Peripheral type benzodiazepine receptors in
human and guinea pig lung: characterization and autoradiographic
mapping. J Pharmacol Exp Ther 252, 880 – 885.
Mantione, C. R., Goldman, M. E., Martin, B., Bolger, G. T., Lueddens, H. W.,
Paul, S. M., et al. (1988). Purification and characterization of an
endogenous protein modulator of radioligand binding to ‘‘peripheral-type’’
benzodiazepine receptors and dihydropyridine Ca2+-channel antagonist
binding sites. Biochem Pharmacol 37, 339 – 347.
Marino, F., Cattaneo, S., Cosentino, M., Rasini, E., Di Grazia, L., Fietta, A. M.,
et al. (2001). Diazepam stimulates migration and phagocytosis of human
neutrophils: possible contribution of peripheral-type benzodiazepine recep-
tors and intracellular calcium. Pharmacology 63, 42 – 49.
Marshall, J. S. (2004). Mast-cell responses to pathogens. Nat Rev Immunol 4,
787 – 799.
Martini, C., Chelli, B., Betti, L., Montali, M., Mancuso, M., Giannaccini, G.,
et al. (2002). Peripheral benzodiazepine binding sites in platelets of patients
affected by mitochondrial diseases and large scale mitochondrial DNA
rearrangements. Mol Med 8, 841 – 846.
Massoco, C., & Palermo-Neto, J. (2003). Effects of midazolam on equine
innate immune response: a flow cytometric study. Vet Immunol Immuno-
pathol 95, 11 – 19.
McEnery, M. W., Snowman, A. M., Trifiletti, R. R., & Snyder, S. H. (1992).
Isolation of the mitochondrial benzodiazepine receptor: association with the
voltage-dependent anion channel and the adenine nucleotide carrier. Proc
Natl Acad Sci U S A 89, 3170 – 3174.
Mercer, K. A., Weizman, R., & Gavish, M. (1992). Ontogenesis of peripheral
benzodiazepine receptors: demonstration of selective up-regulation in rat
testis as a function of maturation. J Recept Res 12, 413 – 425.
Mestre, M., Belin, C., Uzan, A., Renault, C., Dubroeucq, M. C., Gueremy,
C., et al. (1986). Modulation of voltage-operated, but not receptor-
operated, calcium channels in the rabbit aorta by PK 11195, an antagonist
of peripheral-type benzodiazepine receptors. J Cardiovasc Pharmacol 8,
729 – 734.
Mihara, S., & Fujimoto, M. (1989). High-affinity binding sites for PK 11195, but
not for RO5-4864, in porcine aortic smooth muscle. Life Sci 44, 1713 – 1720.
Mihara, S., Sakata, T., & Fujimoto, M. (1990). Interaction of lipocortin I
with peripheral-type benzodiazepine binding sites. Eur J Pharmacol 189,
233 – 236.
Miller, L. G., Lee-Paritz, A., Greenblatt, D. J., & Theoharides, T. C. (1988).
High-affinity benzodiazepine binding sites on rat peritoneal mast cells and
RBL-1 cells: binding characteristics and effects on granule secretion.
Pharmacology 36, 52 – 60.
Milner, P., Bodin, P., Guiducci, S., Del Rosso, A., Kahaleh, M. B., Matucci-
Cerinic, M., et al. (2004). Regulation of substance P mRNA expression in
human dermal microvascular endothelial cells. Clin Exp Rheumatol 22 (3
Suppl 33) (Jan – Feb), s24 – s27.
Miyazawa, N., Diksic, M., & Yamamoto, Y. (1995). Chronological study of
peripheral benzodiazepine binding sites in the rat brain stab wounds using
[3H]PK-11195 as a marker for gliosis. Acta Neurochir 137, 207 – 216.
Moreno-Sanchez, R., Bravo, C., Gutierrez, J., Newman, A. H., & Chiang, P. K.
(1991). Release of Ca2+ from heart and kidney mitochondria by peripheral-
type benzodiazepine receptor ligands. Int J Biochem 23, 207 – 213.
Morgan, J., Oseroff, A. R., & Cheney, R. T. (2004). Expression of the
peripheral benzodiazepine receptor is decreased in skin cancers in
comparison with normal skin. Br J Dermatol 151(4) (Oct), 846 – 856.
Mukherjee, S., & Das, S. K. (1989). Subcellular distribution of ‘‘peripheral
type’’ binding sites for [3H]Ro5-4864 in guinea pig lung. Localization to the
mitochondrial inner membrane. J Biol Chem 264, 16713 – 16718.
Myers, R., Manjil, L. G., Frackowiak, R. S., & Cremer, J. E. (1991). [3H]PK
11195 and the localisation of secondary thalamic lesions following focal
ischaemia in rat motor cortex. Neurosci Lett 133, 20 – 24.
Nakamura, K., Fukunishi, I., Nakamoto, Y., Iwahashi, K., & Yoshii, M. (2002).
Peripheral-type benzodiazepine receptors on platelets are correlated with
the degrees of anxiety in normal human subjects. Psychopharmacology
(Berl) 162, 301 – 303.
Nudmamud, S., Siripurkpong, P., Chindaduangratana, C., Harnyuttanakorn, P.,
Lotrakul, P., Laarbboonsarp, W., et al. (2000). Stress, anxiety and peripheral
benzodiazepine receptor mRNA levels in human lymphocytes. Life Sci 67,
2221 – 2231.
Okubo, T., Yoshikawa, R., Chaki, S., Okuyama, S., & Nakazato, A. (2004).
Design, synthesis, and structure-activity relationships of novel tetracyclic
compounds as peripheral benzodiazepine receptor ligands. Bioorg Med
Chem 12, 3569 – 3580.
Okuyama, S., Chaki, S., Yoshikawa, R., Ogawa, S., Suzuki, Y., Okubo, T., et al.
(1999). Neuropharmacological profile of peripheral benzodiazepine recep-
tor agonists, DAA1097 and DAA1106. Life Sci 64, 1455 – 1464.
Papadopoulos, V., Mukhin, A. G., Costa, E., & Krueger, K. E. (1990). The
peripheral-type benzodiazepine receptor is functionally linked to Leydig
cell steroidogenesis. J Biol Chem 265, 3772 – 3779.
Papadopoulos, V., Guarneri, P., Krueger, K. E., Guidotti, A., & Costa, E.
(1992). Pregnenolone biosynthesis in C6 glioma cell mitochondria:
regulation by diazepam-binding inhibitor mitochondrial receptor. Proc Natl
Acad Sci U S A 89, 5118 – 5122.
Papadopoulos, V., Boujrad, N., Ikonomovic, M. D., Ferrara, P., & Vidic, B.
(1994). Topography of the Leydig cell mitochondrial peripheral-type
benzodiazepine receptor. Mol Cell Endocrinol 104, R5 – R9.
Papadopoulos, V., Amri, H., Boujrad, N., Cascio, C., Culty, M., Garnier, M.,
et al. (1997). Peripheral benzodiazepine receptor in cholesterol transport
and steroidogenesis. Steroids 62, 21 – 28.
Papadopoulos, V., Dharmarajan, A. M., Li, H., Culty, M., Lemay, M., &
Sridaran, R. (1999). Mitochondrial peripheral-type benzodiazepine receptor
expression. Correlation with gonadotropin-releasing hormone (GnRH)
agonist-induced apoptosis in the corpus luteum. Biochem Pharmacol 58,
1389 – 1393.
Park, C. H., Lukacs, L. G., Mastropaolo, J., & Deutsch, S. I. (1997). Selective
effects of MAO inhibition on peripheral benzodiazepine receptor binding in
the mouse. Isr J Psychiatry Relat Sci 34, 300 – 307.
Pedigo, N. W., Schoemaker, H., Morelli, M., McDougal, J. N., Malick, J. B.,
Burks, T. F., et al. (1981). Benzodiazepine receptor binding in young,
mature and senescent rat brain and kidney. Neurobiol Aging 2, 83 – 88.
Peterson, C. L., Duerson, K. C., Buckley, A. R., Putnam, C. W., Russell, D. H.,
& Laird Jr., H. E. (1988). Embryological development of peripheral
benzodiazepine binding sites in the chick liver. Proc West Pharmacol Soc
31, 263 – 264.
Pini, S., Martini, C., Abelli, M., Muti, M., Gesi, C., Montali, M., et al. (2005).
Peripheral-type benzodiazepine receptor binding sites in platelets of patients
 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
with panic disorder associated to separation anxiety symptoms. Psycho-
pharmacology (Berl) (Epub ahead of print).
Primofiore, G., Da Settimo, F., Taliani, S., Simorini, F., Patrizi, M. P.,
Novellino, E., et al. (2004). N,N-dialkyl-2-phenylindol-3-ylglyoxylamides.
A new class of potent and selective ligands at the peripheral benzodiazepine
receptor. J Med Chem 47, 1852 – 1855.
Pubill, D., Verdaguer, E., Canudas, A. M., Sureda, F. X., Escubedo, E.,
Camarasa, J., et al. (2001). Orphenadrine prevents 3-nitropropionic acid-
induced neurotoxicity in vitro and in vivo. Br J Pharmacol 132, 693 – 702.
Rao, V. L., Bowen, K. K., Rao, A. M., & Dempsey, R. J. (2001). Up-regulation
of the peripheral-type benzodiazepine receptor expression and
[3H]PK11195 binding in gerbil hippocampus after transient forebrain
ischemia. J Neurosci Res 64, 493 – 500.
Richfield, E. K., Ciliax, B. J., Starosta-Rubinstein, S. R., McKeever, P. E.,
Penney, J. B., & Young, A. B. (1988). Comparison of [14C]-deoxyglucose
metabolism and peripheral benzodiazepine receptor binding in rat C6
glioma. Neurology 38, 1255 – 1262.
Ritsner, M., Modai, I., Gibel, A., Leschiner, S., Silver, H., Tsinovoy, G.,
et al. (2003). Decreased platelet peripheral-type benzodiazepine recep-
tors in persistently violent schizophrenia patients. J Psychiatr Res 37,
549 – 556.
Rocca, P., Bellone, G., Benna, P., Bergamasco, B., Ravizza, L., & Ferrero, P.
(1993). Peripheral-type benzodiazepine receptors and diazepam binding
inhibitor-like immunoreactivity distribution in human peripheral blood
mononuclear cells. Immunopharmacology 25, 163 – 178.
Rocca, P., Beoni, A. M., Eva, C., Ferrero, P., Zanalda, E., & Ravizza, L. (1998).
Peripheral benzodiazepine receptor messenger RNA is decreased in
lymphocytes of generalized anxiety disorder patients. Biol Psychiatry 43,
767 – 773.
Rocca, P., Beoni, A. M., Eva, C., Ferrero, P., Maina, G., Bogetto, F., et al.
(2000). Lymphocyte peripheral benzodiazepine receptor mRNA
decreases in obsessive – compulsive disorder. Eur Neuropsychopharmacol
10, 337 – 340.
Romeo, E., Auta, J., Kozikowski, A. P., Ma, D., Papadopoulos, V., Puia, G.,
et al. (1992). 2-Aryl-3-indoleacetamides (FGIN-1): a new class of potent
and specific ligands for the mitochondrial DBI receptor (MDR). J
Pharmacol Exp Ther 262, 971 – 978.
Ruff, M. R., Pert, C. B., Weber, R. J., Wahl, L. M., Wahl, S. M., & Paul, S. M.
(1985). Benzodiazepine receptor-mediated chemotaxis of human mono-
cytes. Science 229, 1281 – 1283.
Sacerdote, P., Panerai, A. E., Frattola, L., & Ferrarese, C. (1999). Benzodiaz-
epine-induced chemotaxis is impaired in monocytes from patients with
generalized anxiety disorder. Psychoneuroendocrinology 24, 243 – 249.
Salvetti, F., Chelli, B., Gesi, M., Pellegrini, A., Giannaccini, G., Lucacchini, A.,
et al. (2000). Effect of noise exposure on rat cardiac peripheral
benzodiazepine receptors. Life Sci 66, 1165 – 1175.
Santamaria, M., Lanave, C., & Saccone, C. (2004). The evolution of the
adenine nucleotide translocase family. Gene 333, 51 – 59.
Schoemaker, H., Boles, R. G., Horst, W. D., & Yamamura, H. I. (1983).
Specific high-affinity binding sites for [3H]Ro 5-4864 in rat brain and
kidney. J Pharmacol Exp Ther 225, 61 – 69.
Schwartz-Bloom, R. D., McDonough, K. J., Chase, P. J., Chadwick, L. E.,
Inglefield, J. R., & Levin, E. D. (1998). Long-term neuroprotection by
benzodiazepine full versus partial agonists after transient cerebral ischemia
in the gerbil [Erratum in J Cereb Blood Flow Metab, 1998;18:817]. J Cereb
Blood Flow Metab 18, 548 – 558.
Schwartz-Bloom, R. D., Miller, K. A., Evenson, D. A., Crain, B. J., & Nadler,
J. V. (2000). Benzodiazepines protect hippocampal neurons from degener-
ation after transient cerebral ischemia: an ultrastructural study. Neurosci-
ence 98, 471 – 484.
Selleri, S., Bruni, F., Costagli, C., Costanzo, A., Guerrini, G., Ciciani, G., et al.
(2001). 2-Arylpyrazolo[1,5-a]pyrimidin-3-yl acetamides. New potent and
selective peripheral benzodiazepine receptor ligands. Bioorg Med Chem 9,
2661 – 2671.
Selleri, S., Gratteri, P., Costagli, C., Bonaccini, C., Costanzo, A., Melani, F.,
et al. (2005). Insight into as peripheral benzodiazepine receptor ligands:
synthesis, biological evaluation and 3D-QSAR investigation. Bioorg Med
Chem 13, 4821 – 4834.
523
Slobodyansky, E., Guidotti, A., Wambebe, C., Berkovich, A., & Costa, E.
(1989). Isolation and characterization of a rat brain triakontatetraneuropep-
tide, a posttranslational product of diazepam binding inhibitor: specific
action at the Ro 5-4864 recognition site. J Neurochem 53, 1276 – 1284.
Snyder, M. J., & Van Antwerpen, R. (1998). Evidence for a diazepam-binding
inhibitor (DBI) benzodiazepine receptor-like mechanism in ecdysteroido-
genesis by the insect prothoracic gland. Cell Tissue Res 294, 161 – 168.
Snyder, S. H., Verma, A., & Trifiletti, R. R. (1987). The peripheral-type
benzodiazepine receptor: a protein of mitochondrial outer membranes
utilizing porphyrins as endogenous ligands. FASEB J 1, 282 – 288.
Sommer, C. (2004). Serotonin in pain and analgesia: actions in the periphery.
Mol Neurobiol 30, 117 – 125.
Soreni, N., Apter, A., Weizman, A., Don-Tufeled, O., Leschiner, S., Karp, L.,
et al. (1999). Decreased platelet peripheral-type benzodiazepine receptors in
adolescent inpatients with repeated suicide attempts. Biol Psychiatry 46,
484 – 488.
Steinberg, R. A., Symcox, M. M., Sollid, S., & Ogreid, D. (1996). Arginine 210
is not a critical residue for the allosteric interactions mediated by binding of
cyclic AMP to site A of regulatory (RIalpha) subunit of cyclic AMP-
dependent protein kinase. J Biol Chem 271, 27630 – 27636.
Stephenson, D. T., Schober, D. A., Smalstig, E. B., Mincy, R. E., Gehlert, D.
R., & Clemens, J. A. (1995). Peripheral benzodiazepine receptors are
colocalized with activated microglia following transient global forebrain
ischemia in the rat. J Neurosci 15, 5263 – 5274.
Stoebner, P. E., Carayon, P., Penarier, G., Frechin, N., Barneon, G., Casellas, P.,
et al. (1999). The expression of peripheral benzodiazepine receptors in
human skin: the relationship with epidermal cell differentiation. Br J
Dermatol 140, 1010 – 1016.
Suzuki-Nishimura, T., Sano, T., & Uchida, M. K. (1989). Effects of
benzodiazepines on serotonin release from rat mast cells. Eur J Pharmacol
167, 75 – 85.
Syapin, P. J., & Skolnick, P. (1979). Characterization of benzodiazepine binding
sites in cultured cells of neural origin. J Neurochem 32, 1047 – 1051.
Taniguchi, T., Wang, J. K., & Spector, S. (1980). Properties of [3H] diazepam
binding to rat peritoneal mast cells. Life Sci 27, 171 – 178.
Tanne, Z., Weizman, R., Karp, L., Katz, Y., Tyano, S., & Gavish, M. (1987).
Lack of effect of 28 days of neuroleptic treatment on platelet benzodiaz-
epine binding sites in schizophrenics. Neuropsychobiology 17, 121 – 123.
Thinnes, F. P. (1992). Evidence for extra-mitochondrial localization of the
VDAC/porin channel in eukaryotic cells. J Bioenerg Biomembranes 24,
71 – 75.
Trapani, G., Franco, M., Latrofa, A., Ricciardi, L., Carotti, A., Serra, M., et al.
(1999). Novel 2-phenylimidazo[1,2-a]pyridine derivatives as potent and
selective ligands for peripheral benzodiazepine receptors: synthesis, binding
affinity, and in vivo studies. J Med Chem 42, 3934 – 3941.
Tsujimoto, Y., & Shimizu, S. (2000). VDAC regulation by the Bcl-2 family of
proteins. Cell Death Differ 7, 1174 – 1181.
Van Hinsbergh, V. W., Koolwijk, P., & Hanemaaijer, R. (1997). Role of fibrin
and plasminogen activators in repair-associated angiogenesis: in vitro
studies with human endothelial cells. EXS 79, 391 – 411.
Veenman, L., & Gavish, M. (2000). Peripheral-type benzodiazepine receptors:
their implication in brain disease. Drug Dev Res 50, 355 – 370.
Veenman, L., Leschiner, S., Spanier, I., Weisinger, G., Weizman, A., & Gavish,
M. (2002). PK11195 attenuates kainic acid-induced seizures and alterations
in peripheral-type benzodiazepine receptor (PBR) components in the rat
brain. J Neurochem 80, 917 – 927.
Veenman, L., Levin, E., Weisinger, G., Leschiner, S., Spanier, I., Snyder, S.
H., et al. (2004). Peripheral-type benzodiazepine receptor density and
in vitro tumorigenicity of glioma cell lines. Biochem Pharmacol 68,
689 – 698.
Verma, A., & Snyder, S. H. (1988). Characterization of porphyrin interac-
tions with peripheral type benzodiazepine receptors. Mol Pharmacol 34,
800 – 805.
Verma, A., & Snyder, S. H. (1989). Peripheral type benzodiazepine receptors.
Ann Rev Pharmacol Toxicol 29, 307 – 322.
Verma, A., Nye, J. S., & Snyder, S. H. (1987). Porphyrins are endogenous
ligands for the mitochondrial (peripheral-type) benzodiazepine receptor.
Proc Natl Acad Sci U S A 84, 2256 – 6220.
524 L. Veenman, M. Gavish / Pharmacology & Therapeutics 110 (2006) 503 – 524
Verrier, F., Mignotte, B., Jan, G., & Brenner, C. (2003). Study of PTPC
composition during apoptosis for identification of viral protein target. Ann
NY Acad Sci 1010, 126 – 142.
Versijpt, J., Dumont, F., Thierens, H., Jansen, H., De Vos, F., Slegers, G., et al.
(2000). Biodistribution and dosimetry of [123I]iodo-PK 11195: a potential
agent for SPET imaging of the peripheral benzodiazepine receptor. Eur J
Nucl Med 27, 1326 – 1333.
Vin, V., Leducq, N., Bono, F., & Herbert, J. M. (2003). Binding characteristics
of SSR180575, a potent and selective peripheral benzodiazepine ligand.
Biochem Biophys Res Commun 310, 785 – 790.
Wang, J. K. T., Morgan, J. I., & Spector, S. (1984). Benzodiazepines that bind
at peripheral sites inhibit cell proliferation. Proc Natl Acad Sci U S A 81,
753 – 756.
Weintraub, W. H., Cleveland-Wolfe, P., & Fewtrell, C. (1994). Paracrine Ca2+
signaling in vitro: serotonin-mediated cell-cell communication in mast
cell/smooth muscle cocultures. J Cell Physiol 160, 389 – 399.
Weizman, R., Tanne, Z., Karp, L., Tyano, S., & Gavish, M. (1986). Peripheral-
type benzodiazepine-binding sites in platelets of schizophrenics with and
without tardive dyskinesia. Life Sci 39, 549 – 555.
Weizman, R., Tanne, Z., Granek, M., Karp, L., Golomb, M., Tyano, S., et al.
(1987). Peripheral benzodiazepine binding sites on platelet membranes are
increased during diazepam treatment of anxious patients. Eur J Pharmacol
138, 289 – 292.
Weizman, A., Tanne, Z., Karp, L., Martfeld, Y., Tyano, S., & Gavish, M.
(1987). Carbamazepine up-regulates the binding of [3H]PK 11195 to
platelets of epileptic patients. Eur J Pharmacol 141, 471 – 474.
Weizman, A., Amiri, Z., Katz, Y., Snyder, S. H., & Gavish, M. (1992).
Testosterone prevents castration-induced reduction in peripheral benzodi-
azepine receptors in Cowper’s gland and adrenal. Brain Res 572, 72 – 75.
Weizman, R., Laor, N., Karp, L., Dagan, E., Reiss, A., Dar, D. E., et al. (1994).
Alteration of platelet benzodiazepine receptors by stress of war. Am J
Psychiatry 151, 766 – 767.
Weizman, A., Burgin, R., Harel, Y., Karp, L., & Gavish, M. (1995). Platelet
peripheral-type benzodiazepine receptor in major depression. J Affect
Disord 33, 257 – 261.
Weizman, R., Karp, L., Dar, D. E., Butin, B., Hermesh, H., Munitz, H., et al.
(1996). Electroconvulsive therapy down-regulates platelet peripheral-type
benzodiazepine receptors in medication-resistant major depressed patients.
Biol Psychiatry 40, 221 – 224.
Weizman, R., Dagan, E., Snyder, S. H., & Gavish, M. (1997a). Impact of
pregnancy and lactation on GABAA receptor and central-type and
peripheral-type benzodiazepine receptors. Brain Res 752, 307 – 314.
Weizman, R., Leschiner, S., Schlegel, W., & Gavish, M. (1997b). Peripheral-
type benzodiazepine receptor ligands and serum steroid hormones. Brain
Res 772, 203 – 208.
Woods, M. J., & Williams, D. C. (1996). Multiple forms and locations for
the peripheral-type benzodiazepine receptor. Biochem Pharmacol 52,
1805 – 1814.
Wojta, J., Huber, K., & Valent, P. (2003 – 2004). New aspects in thrombotic
research: complement induced switch in mast cells from a profibrinolytic to
a prothrombotic phenotype. Pathophysiol Haemost Thromb 33, 438 – 441.
Yong, L. C. (1997). The mast cell: origin, morphology, distribution, and
function. Exp Toxicol Pathol 49, 409 – 424.
Zhang, M. R., Kida, T., Noguchi, J., Furutsuka, K., Maeda, J., Suhara, T., et al.
(2003). [11C]DAA1106: radiosynthesis and in vivo binding to peripheral
benzodiazepine receptors in mouse brain. Nucl Med Biol 30, 513 – 519.
Zhang, M. R., Maeda, J., Ogawa, M., Noguchi, J., Ito, T., Yoshida, Y., et al.
(2004). Development of a new radioligand, N-(5-fluoro-2-phenoxyphenyl)-
N-(2-[18F]fluoroethyl-5-methoxybenzyl)acetamide, for pet imaging of
peripheral benzodiazepine receptor in primate brain. J Med Chem 47,
2228 – 2235.
Zhang, M. R., Maeda, J., Ito, T., Okauchi, T., Ogawa, M., Noguchi, J., et al.
(2005). Synthesis and evaluation of N-(5-fluoro-2-phenoxyphenyl)-N-(2-
[(18)F]fluoromethoxy-d(2)-5-methoxybenzyl)acetamide: a deuterium-sub-
stituted radioligand for peripheral benzodiazepine receptor. Bioorg Med
Chem 13, 1811 – 1818.
Zilz, A., Li, H., Castello, R., Papadopoulos, V., & Widmaier, E. P. (1999).
Developmental expression of the peripheral-type benzodiazepine receptor
and the advent of steroidogenesis in rat adrenal glands. Endocrinology 140,
859 – 864.
Zisterer, D. M., Gorman, A. M., Williams, D. C., & Murphy, M. P. (1992). The
effects of the peripheral-type benzodiazepine acceptor ligands, Ro 5-4864
and PK 11195, on mitochondrial respiration. Methods Find Exp Clin
Pharmacol 14, 85 – 90.