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258 Journal of Investigative Dermatology (2006), Volume 126 2006 The Society for Investigative Dermatology
Apoptosis in the Hair Follicle
Natalia V. Botchkareva
1
, Gurpreet Ahluwalia
1
and Douglas Shander
1
A p o p to si s p lays an i m p o rtan t ro le i n m an y p h ysi o lo g i cal p ro ce sse s, ran g i n g fro m m o rp h o g e n e ti c e ve n ts to ad u lt
ti ssu e h o m e o stasi s, an d d e fe cts i n i ts re g u lati o n co n tri b u te to m an y d i so rd e rs. H e re we re vi e w m o le cu lar m e ch a-
n i sm s o f ap o p to si s i n th e h ai r fo lli cle H F) , wh o se cycli cal g ro wth p atte rn i s re p e ate d ly i n te rru p te d b y ap o p -
to si s-d ri ve n i n vo lu ti o n catag e n ) . We re vi e w th e co m m o n m e ch an i sm s u n d e rlyi n g ap o p to si s i n th e H F d u ri n g
catag e n , as we ll as d i ffe re n ce s i n th e re g u lati o n o f ap o p to si s b e twe e n d i sti n ct H F ce ll p o p u lati o n s. A n o ve rvi e w i s
p ro vi d e d o n th e e xp re ssi o n an d fu n cti o n o f m o le cu le s i n vo lve d i n th e co n tro l o f vari o u s p h ase s o f th e ap o p to ti c
p ro ce ss d u ri n g catag e n .
Journal of Investigative Dermatology (2006) 126, 258264. doi:10.1038/sj.jid.5700007
Apoptosis is a process of eliminating
cells that have fulfilled their biological
function during development and tissue
homeostasis in multicellular organisms.
During the last decade, substantial prog-
ress was achieved in the delineation of
molecular mechanisms involved in the
realization of apoptosis. As an energy-
demanding process, apoptosis is initi-
ated by a variety of stimuli, including a
withdrawal of growth factors, loss of cell
adhesion, stimulation of proapoptotic
receptors, or DNA damage (reviewed
by Afford and Randhawa, 2000).
The hair follicle (HF) is a cutaneous
organ that shows cyclic activity in post-
natal life and transits through periods
of active hair growth (anagen), apop-
tosis-driven involution (catagen), hair
shedding (exogen), and relative resting
(telogen) (Paus and Cotsarelis, 1999;
Cotsarelis and Millar, 2001; Stenn and
Paus, 2001). During HF growth and
hair production, the activity of factors
promoting the proliferation, differen-
tiation, and survival predominates,
whereas HF regression is characterized
by the activation of a variety of signal-
ing pathways that induce apoptosis in
HF cells (Lindner et al., 1997; Soma et
al., 1998; Stenn and Paus, 2001). This
review focuses on the molecular mech-
anisms of apoptosis in the HF during its
physiological involution; mechanisms
of apoptosis underlying the distinct hair
loss conditions have been reviewed
elsewhere (Botchkarev, 2003; Cotsarelis
and Millar, 2001; Hendrix et al., 2005).
Apoptosis As a Multi-Step Program of
Cell Death
Morphologically, apoptosis is charac-
terized by cell shrinkage, membrane
blebbing, nuclear condensation, and
cellular fragmentation, followed by
the formation and the phagocytosis of
apoptotic bodies. The process of apop-
tosis can be divided into four stages:
initiation, intracellular response,
cell fragmentation, and phagocytosis
(reviewed by Afford and Randhawa,
2000).
Stages of Initiation and Intracellular
Response of Apoptosis
The extrinsic apoptotic pathway is acti-
vated by binding of specific ligands
to the members of the death domain
family of membrane receptors (Curtin
and Cotter, 2003) (Figure 1). All death
receptors have an extracellular cys-
teine-rich domain that is required for
ligand binding and an intracellular
domain essential for signal transduction
(Peter and Krammer, 2003).
Signaling via death receptors results
in the recruitment of several intracel-
lular adapter molecules specific for
each apoptotic receptor (Fas-associ-
ated death domain protein, tumor
necrosis factor (TNF) receptor 1asso-
ciated death domain, receptor-inter-
acting protein 1, and so on; see Figure
1), all of which contain the death
effector domains required for the
interaction with similar domains of pro-
caspase-8 and procaspase-10 (Curtin
and Cotter, 2003; Roux and Barker,
2002). Activation of caspase-8 and cas-
pase-10 results in their recruitment into
the death-inducing signaling complex,
followed by activation of caspase-3
along the common apoptotic pathway
(Peter and Krammer, 2003). Caspase-8
may also activate the cytoplasmic pro-
tein Bid, which, after its translocation
to the mitochondria, induces release of
cytochrome c, thus linking the extrin-
sic and intrinsic apoptotic pathways
(Schultz and Harrington, 2003).
The intrinsic pathway of apoptosis
is activated by a variety of intracellular
events that lead to the release of cyto-
chrome c from the mitochondria. This
process is controlled by proteins of the
Bcl-2 family, which are divided into
two groups: inhibitors of apoptosis (for
example, Bcl-2, Bcl-x
L
, and Mcl-1) and
promoters of apoptosis (for example,
Bax, Bak, Bok, Bcl-x
S
, Bik, Bim, Bad,
and Bid) (van Gurp et al., 2003). Bim,
Bad, and Bid may act as sensors for
1
Gillette Technology Center, Needham, Massachusetts, USA
Correspondence: Natalia V. Botchkareva or Douglas Shander, Gillette Technology Center, 37 A Street, Needham, Massachusetts 02492, USA.
E-mail: natasha_botchkareva@gillette.com or doug_shander@gillette.com
Abbreviations: HF, hair follicle; IAP, inhibitor of apoptosis protein; KC, keratinocyte; TGF-, transforming growth factor-; TNF-, tumor necrosis factor-
Manuscript received 5 July 2005; revised 16 August 2005; accepted for publication 2 September 2005
NV Botchkareva et al.
Hair follicle apoptosis
www.jidonline.org 259
cellular integrity and the growth factor
supply. Bim serves as a sensor of cyto-
skeleton integrity, and its proapoptotic
activity is regulated by interaction with
the dynein motor complex (Puthalakath
et al., 1999). Bad operates as a sensor
for growth factor withdrawal, as depriva-
tion of survival growth factors induces
Bad dephosphorylation, which results in
apoptosis (Chiang et al., 2001). The cyto-
plasmic protein Bid may be cleaved by
caspase-8, granzyme B, and cathepsin
into a truncated isoform (tBid), which,
after translocation to the mitochondria,
promotes cytochrome c release and
apoptosis (van Gurp et al., 2003).
After its release into the cytoplasm,
cytochrome c recruits the caspase adapt-
er molecule Apaf-1 and the apoptosis ini-
tiator enzyme procaspase-9, which toge-
ther form a holoenzyme complex called
an apoptosome (van Gurp et al., 2003).
Apoptosomes promote the formation of
the active form of caspase-3 along the
common pathway of apoptosis (Figure
1). Caspase-3 activation may be inhibited
by members of the inhibitor of apoptosis
protein (IAP) family, which are located in
the cytosol and prevent the activation of
procaspases. However, during apoptosis
the inhibitory activity of IAPs is neutral-
ized by Smac/DIABLO protein, which is
released from the mitochondria and is
capable of sequestering IAPs (van Gurp
et al., 2003; Riedl and Shi, 2004).
Stage of Cell Fragmentation and
Phagocytosis of Apoptotic Cells
Both extrinsic and intrinsic apoptotic
pathways result in the activation of cas-
pase-3, which is believed to be the key
proteolytic enzyme capable of cleaving
a large variety of intracellular substrates.
These include structural and signaling
proteins, transcription factors, and regu-
lators of DNA repair and RNA metabo-
lism (Shi, 2002). In addition, caspase-3
cleaves caspase-activated deoxyri-
bonuclease, which translates into the
nucleus and initiates DNA cleavage
(van Gurp et al., 2003). These processes
result in the appearance of the degraded
chromatin fragments, lysosomes, mito-
chondria, and other degenerated cell
organelles covered by cell membrane,
which are morphologically distinguish-
able as apoptotic bodies (Afford and
Randhawa, 2000; Shi, 2002).
During the final steps of apoptosis,
cellular, and nuclear fragmentation is
also accompanied by changes in the
cell membrane and by the translocation
of phosphatidylserine from the inner to
the outer portion of the cell membrane,
leading to its recognition by neighbor-
ing cells, macrophages, or dendritic
cells and then to its phagocytosis
(Fadok, 1999; Afford and Randhawa,
2000; Schultz and Harrington, 2003).
CD14, CD36, CD68, the class A sca-
venger receptors and the vitronectin
receptor expressed by macrophages
are all capable of interaction with sev-
eral molecular determinants expressed
on apoptotic cells, including anionic
phospholipids, phosphatidylserine, and
ICAM-3 (Afford and Randhawa, 2000).
In addition, serum proteins such as
2-glycoprotein and the complement
component C1q can bind to apoptotic
cells and enhance their uptake by mac-
rophages (Kagan et al., 2003).
Physiological Hair Follicle Regression
(Catagen) As a Model of Apoptosis-Driven
and Fully Reversible Organ Involution
Although catagen is often thought of
as a regressive event, it is an exqui-
sitely orchestrated energy-requir-
ing remodeling process, whose pro-
gression assures renewal of further
generation of HFs. Catagen was first
characterized in detail by Kligman
and by Straile et al. (Kligman, 1959;
Straile et al., 1961). It lasts about
23 weeks in humans and 3 days in
mice (Kligman, 1959; Parakkal, 1970;
Straile et al., 1961). Morphologically
and functionally, catagen is divided
into eight substages, each character-
ized by distinct patterns of apoptosis
(Mller-Rver et al., 2001) (Figure 2a).
During the murine hair cycle, apop-
totic cells visualized by TUNEL staining
first appear in the melanogenic area of
the late-anagen HF (Tobin et al., 1998).
In early-catagen HFs, apoptotic cells are
seen in the HF matrix; later, TUNEL+
cells are seen in the outer and inner
root sheaths (mid- and late catagen)
and are then abundant in the epithe-
lial strand (late catagen) (Lindner et al.,
1997; Matsuo et al., 1998). In advanced
catagen stages, TUNEL+ cells are also
present in the club hair and in the bulge
(Lindner et al., 1997; Ito et al., 2004).
However, apoptotic cells are never seen
in the dermal papilla of catagen HFs
in humans and mice under physiologi-
cal conditions (Lindner et al., 1997;
Intracellular Response Cell Fragmentation
Neurotrophins
TNF
FasL, DcR3
TL1A
Trail
Membrane of the intact ce
Mitochondria
Nucleus
Initiation Stage Intracellular Response Cell Fragmentation Phagocytosis
Apoptotic
ligands
Neurotrophins
TNF-
FasL, DcR3
TL1A DR3 (TRAMP/Apo3) TRADD, RIP, TRAF
TRAIL DR5 (TRAILR2/TRICK2) TRAF, FADD
DR4 (TRAILR1/Apo2) FADD
DcR1, DcR2
? DR6 TRADD, TRAF
Fas (CD95/Apo1) FAF1, DAXX, FADD
TNFR-1 TRADD, RIP, TRAF
p75NTR NRAGE, NRIF-1/2, NADE
Lysosomal
defects
Caspase-8
activation
Smac/
DIABLO
Endonuclease G
AIF
Apoptosome
Cyt c
Apaf-1
Procaspase-9
Death receptors and adapter molecules
Jnk
Bad
IAP
Bax
Bak
Bcl-2
Bcl-x
L
Bim
Bmf
c-Jun
Cathepsins
Bid tBid
Growth
factor
withdrawal
Loss of cytoskeleton
integrity
Caspase-
activated
DNAse
Decrease of
Akt kinase
activity
Caspase-3
activation
Proteolysis of
cytoplasmic
substrates
Phagocyte receptors
(CD14, CD36, CD68)
Phosphatidyl
serine
residues
Phosphatidylserine
residues
Proteolysis of
cytoplasmic
substrates
Cyt c
Endo G
AIF
Smac
Transcription
of apoptotic
regulators
(Bax)
DNA
fragmentation
Loss of cell
adhesion
Dynein motor
complex - Bim/Bmf
Membrane of the intact cell
Membrane of
apoptotic cell
Membrane of
phagocyte
Figure 1. Molecular mechanisms of the distinct phases of the apoptotic process. Scheme demonstrates
integration of the molecules involved in the control of various phases of apoptosis (initiation,
intracellular response, cell fragmentation, phagocytosis). AIF, apoptosis-inducing factor; Akt, protein
kinase B; Apaf-1, apoptotic peptidase activating factor 1 Cyt c, cytochrome c; Endo G, endonuclease G;
Jnk, c-Jun N-terminal kinase.
NV Botchkareva et al.
Hair follicle apoptosis
260 Journal of Investigative Dermatology (2006), Volume 126
Matsuo et al., 1998; Soma et al., 1998).
Therefore, the spatio-temporal distribu-
tion of TUNEL+ cells in the HF during
catagen may be considered as a wave
starting from the melanogenic area (late
anagen), propagating to the hair matrix
(early catagen) and then to the proxi-
mal/central outer and inner root sheaths
and hair shaft (mid- and late catagen).
Distinct cell populations in the HF
possess differential susceptibility to
apoptosis. The most susceptible to
apoptosis are the majority of the follic-
ular epithelial cells and melanocytes,
whereas dermal papilla fibroblasts,
and some of the keratinocytes (KCs)
and melanocytes selected for survival,
are resistant to apoptosis (Lindner et
al., 1997; Seiberg et al., 1995; Tobin
et al., 1998). Apoptotic cells in the HF
are phagocytosed by macrophages and
by neighboring epithelial cells, which
fill the space left by apoptotic cells
(the apoptotic force hypothesis intro-
duced by K. Stenn (Stenn and Paus,
2001)). Loss of cell adhesion between
KCs may also be considered a catagen-
promoting factor, as ICAM-1-deficient
mice show significant catagen accel-
eration (Mller-Rver et al., 2000a).
This loss of cell adhesion leads to a
progressive reduction in the size of the
proximal hair bulb (early and mid-
catagen) and to rapid shortening of the
outer and inner root sheaths and hair
shaft (mid- and late catagen).
Catagen is also characterized by the
formation of a specialized structure in
the HF, the club hair, that connects the
proximal part of the hair shaft with sur-
rounding HF epithelium and anchors
hair in the telogen HF (Parakkal, 1970;
Pinkus et al., 1981). During catagen,
the dermal papilla is transformed into a
cluster of quiescent cells closely adja-
cent to the regressing HF epithelium,
which moves from the subcutis to the
dermis/subcutis border to contact the
distal portion of the HF epithelium,
including the secondary hair germ and
bulge. In the hairless gene mutation, the
disintegration of the dermal papilla and
the HF epithelium during catagen leads
to the loss of the capacity of the HF to
reenter anagen (Ahmad et al., 1998;
Panteleyev et al., 1999).
Molecular Mechanisms of Apoptosis in
the Hair Follicle During Catagen
The physiological involution of the HF
may be triggered by a variety of stimu-
li, including signaling via death recep-
tors, and by the withdrawal of growth
factors that maintain cell proliferation
and differentiation in the anagen HF.
Accumulating evidence suggests that
apoptosis in every distinct HF compart-
ment is regulated differently (Figure 3).
Apoptosis in the follicular melano-
cytes. Recent data based on monitoring
of the fate of melanocytes during cata-
gen in Trp2-LacZ transgenic mice sug-
gest that distinct subsets of melanocytes
(located in the bulge, in the outer root
sheath, and above the dermal papilla;
Botchkareva et al., 2001) show differ-
ent sensitivities to apoptosis (Sharov et
al., 2005). Specifically, apoptosis occurs
only in differentiated melanocytes that
are located above the dermal papilla
and express the Fas receptor, whereas
the other subpopulations, which express
the c-Kit receptor and Bcl-2, most likely
survive catagen (Sharov et al., 2005).
Thus, Fas signaling may contribute to
melanocyte apoptosis during catagen,
similar to that seen after exposure of HFs
to chemotherapy (Sharov et al., 2003).
Stem cell factor/c-Kit and Bcl-2 are
two important candidate molecules
that may promote melanocyte survival
during catagen. Constitutive stem cell
factor/c-Kit ablation or administration
of c-Kit-neutralizing antibody leads to
apoptosis and the disappearance of
a f
Anagen Telogen
*
*
d e b c
*
Catagen II Catagen V
Figure 2. Catagen development in the human HF. (a) Alterations in HF morphology at distinct stages of anagen-catagen-telogen transition. During anagen-
catagen transition (Catagen II), the size of the hair matrix (arrow) and dermal papilla (arrowhead) is reduced. During the advanced catagen stages (Catagen
V), the dermal papilla is condensed and transformed into a cluster of quiescent cells (arrowhead), and the epithelial strand (arrow) and club hair (asterisk)
are formed. In telogen (right), HF length is substantially reduced, and the HF contains two specialized structures, the secondary hair germ (arrowhead) and
club hair (asterisk). (b), (c) Bax. In anagen, weak Bax expression is seen in the central inner root sheath (b, arrow). In catagen, Bax expression appears in the
regressing hair matrix, the inner root sheath, and the connective tissue sheath (c, large arrowhead, arrow, and small arrowhead, respectively). (d), (e) Caspase-
8. In anagen, caspase-8 expression is restricted to the perifollicular connective tissue sheath (d, arrows). In early catagen, in addition to the connective tissue
sheath, caspase-8 appears in central and distal parts of the outer root sheath (e, arrow and arrowhead, respectively). (f) TUNEL (green) plus Ki67 (red). Laser
treatment promotes catagen development accompanied by massive apoptosis in the HF: TUNEL+ cells are located in the regressing epithelium (arrow), club
hair (arrowhead), and dermal papilla (asterisk).
NV Botchkareva et al.
Hair follicle apoptosis
www.jidonline.org 261
melanoblasts in mouse embryos (Ito et
al., 1999). Bcl-2 specifically protects the
melanocyte stem cells in the HF from
apoptosis, and Bcl-2 deficiency results
in their apoptotic elimination and pre-
mature hair graying (Veis et al., 1993;
Nishimura et al., 2005). However,
cross-talk between the stem cell fac-
tor/c-Kit pathway and mitochondrial
antiapoptotic proteins in the control of
melanocyte survival has yet to be fur-
ther clarified.
Apoptosis in the hair matrix kerati-
nocytes. Data obtained from murine
and human HFs suggest that apop-
tosis in hair matrix KCs is controlled
by extrinsic and intrinsic pathways.
The p55 subunit of the TNF recep-
tor may be involved in the extrinsic
apoptotic pathway, as it is expressed
in the hair matrix during catagen, and
TNF- treatment results in an increase
of TUNEL+ cells in the hair matrix
(Lindner et al., 1997; Ruckert et al.,
2000). Also, TNF- promotes anagen-
catagen transition in human HFs in
vitro (Hoffmann et al., 1996).
Apoptosis in hair matrix KCs is also
accompanied by the activation of
intrinsic apoptotic pathways. During
catagen in mice, the Bcl-2/Bax ratio in
hair matrix KCs is markedly decreased,
compared with anagen levels (Figure
2b and c) (Lindner et al., 1997). The
Survivin protein, which belongs to the
IAP family, is prominently expressed in
the proliferating cells of the hair matrix,
whereas its expression is progressively
decreased during catagen (Botchkareva
et al., 2005).
The p53 transcription factor also
plays an important role in regulation
of apoptosis in hair matrix KCs: cata-
gen HFs of p53-null mice contain
fewer apoptotic cells and show sig-
nificantly retarded catagen progres-
sion, compared with catagen HFs of
wild-type mice (Botchkarev et al.,
2001). p53 may promote the intrinsic
apoptotic pathway in hair matrix by
regulating the expression of the mito-
chondrial protein Bax, as such expres-
sion is increased in the hair matrix
during catagen (Lindner et al., 1997;
Botchkarev et al., 2001).
The hairless transcription factor also
serves as an important regulator of
apoptosis in the HF: in hairless mice,
apoptosis in hair matrix KCs is strong-
ly increased, leading to a premature
switch from anagen to catagen and
eventually resulting in disintegration
of the dermal papilla from the follicu-
lar epithelium and permanent hair loss
(Ahmad et al., 1998; Panteleyev et al.,
1999; Panteleyev et al., 2000). It was
recently shown that hairless protein
serves as a transcriptional co-repressor
for the vitamin D receptor (Hsieh et al.,
2003; Potter et al., 2001); this suggests
that interactions between hairless pro-
tein and vitamin D receptor might con-
tribute to apoptosis in HF KCs.
Interestingly, apoptosis in the hair
matrix is strongly increased in keratin
17 knockout mice (McGowan et al.,
2002); this provides the first evidence
that ablation of a structural protein
may cause apoptosis in the HF KCs. It
remains to be determined, however,
whether keratin 17 deletion results in
direct activation of the intrinsic apoptot-
ic pathway, employing Bim as a sensor
of cytoskeleton integrity (Puthalakath
et al., 1999), or whether other mecha-
nisms are involved in this process.
Effector mechanisms of apoptosis in
the murine and human HFs are most
likely mediated by caspase-1, caspase-
3, caspase-4, and caspase-7, which are
highly expressed in the hair matrix dur-
ing catagen and may be involved in the
cell fragmentation process (Lindner et
al., 1997; Soma et al., 1998).
Apoptosis in the outer root sheath.
Before or during catagen, outer root
sheath KCs produce several important
catagen-promoting secreted molecules:
fibroblast growth factor-5 short isoform,
neurotrophins, transforming growth
factor-1/2 (TGF-1/2), IGF bind-
ing protein 3, and thrombospondin-1
(Botchkarev et al., 2001; Botchkarev
et al., 2004; Suzuki et al., 1998; Yano
et al., 2003). Overexpression of one of
the antiapoptotic mitochondrial pro-
teins Bcl-2 and Bcl-x
L
in the outer root
sheath KCs leads to premature termina-
tion of anagen and catagen accelera-
tion, whereas fibroblast growth factor-5
deficiency rescues these effects (Pena et
al., 1999; Mller-Rver et al., 2000b.
This suggests that the elevated levels
of Bcl-2 and Bcl-x
L
in the outer root
sheath stimulate survival and/or activ-
ity of cells that promote HF anagen-
catagen transition.
Along with the secretion of cata-
gen stimulatory molecules, outer root
sheath KCs express apoptotic death
receptors such as Fas, p55 TNF recep-
tor, and p75 neurotrophin receptor, as
well as TGF-1/2 receptors (Lindner
et al., 1997; Paus et al., 1997; Foitzik
et al., 2000). This suggests that outer
root sheath cells are also the targets
for active elimination during catagen.
The central outer root sheath of human
catagen HFs displays high expression
of caspase-8 (Figure 2d and 2e), which
links death receptor signaling with
the activation of caspase-3 (Peter and
Krammer, 2003). Outer root sheath KCs
expressing p75NTR and TGF- recep-
tors become TUNEL+ during catagen
(Botchkarev et al., 2000; Foitzik et al.,
2000). In contrast to corresponding
wild-type mice, p75NTR and TGF-1
knockout mice display significant cata-
gen retardation associated with the
decline of apoptotic cells in the regress-
ing outer root sheath (Botchkarev et al.,
2000; Foitzik et al., 2000). Thus, by
stimulating apoptosis, neurotrophins
and TGF-1/2 may promote shortening
of the outer root sheath in catagen HFs.
These data are consistent with results
that show potent catagen-promoting
activities of neurotrophins and TGF-2
in human HFs (Soma et al., 2002; Tsuji
et al., 2003; Peters et al., 2005).
Outer root sheath
Fas
p75NTR
TGF- -1/2 receptors
p53
Inner root sheath
L-cathepsin
hairless
GDNF receptors
Fas Hair matrix
c-Kit
Bcl-2
hairless
Survivin
Bax
p53
TNF-/p55TNFR
Melanocytes
c-Kit
Bcl-2
Fas
Dermal papilla
Bcl-2
Figure 3. Molecular mechanisms of apoptosis
control in the distinct HF compartments. Scheme
demonstrates the expression pattern of anti- and
proapoptotic molecules (shown in brown and
black, respectively) in the HF. GDNF, glial cell
linederived neurotrophic factor.
NV Botchkareva et al.
Hair follicle apoptosis
262 Journal of Investigative Dermatology (2006), Volume 126
Apoptosis in the inner root sheath
and the hair shaft. During catagen,
inner root sheath KCs show strong
upregulation of the Fas/apo receptor
(Lindner et al., 1997), implicating its
involvement in apoptosis. Proteolytic
enzyme L-cathepsin and hairless may
also contribute to apoptosis in the
inner root sheath, as both L-cathepsin-
deficient and hairless-deficient mice
show retarded inner root sheath short-
ening and decreased apoptosis during
catagen (Panteleyev et al., 2000; Tobin
et al., 2002). Antiapoptotic effects in
the inner root sheath may be mediated
by glial cell linederived neurotrophic
factor and neurturin; their correspond-
ing receptors, GFR1 and GFR2, are
expressed in the inner root sheath dur-
ing catagen, and GFR1- and GFR2-
deficient mice show significant catagen
acceleration, compared with wild-type
controls (Botchkareva et al., 2000).
Less is known about the mechanisms
that control apoptosis in hair shaft KCs.
In catagen, hair shaft KCs do not express
apoptotic receptors (Lindner et al.,
1997), although single TUNEL+ cells
could be detected (Soma et al., 1998).
During mid-catagen and late catagen,
TUNEL+ cells become abundant in the
proximal end of the hair shaft, that is, in
the area of trichilemmal keratinization
and club hair formation (Lindner et al.,
1997; Matsuo et al., 1998). However, as
TUNEL staining is not always a marker
of apoptotic cells (Magerl et al., 2001),
it is difficult to conclude whether apop-
tosis plays a role in club hair formation.
Clearance of the apoptotic cells from
the HF during catagen. Little is known
about the mechanisms that control
clearance of the apoptotic cells from
the HF during catagen. As was shown
in other models of biological regres-
sion (involution of the mammary gland
or thymic epithelium), apoptotic cells
are eliminated by macrophages or by
neighboring epithelial cells (reviewed
by Fadok, 1999). Dendritic cells that
are present in human anagen HFs in
distal HF epithelium and express C1q
may be closely involved in the rec-
ognition and phagocytosis of apop-
totic cells (Christoph et al., 2000).
Apoptotic cells may also be eliminated
by macrophages that are located in
the connective tissue sheath and that
express CD68 (Christoph et al., 2000).
Interestingly, the molecules that stimu-
late catagen development (interleukin
1, TNF-, TGF-1) are also known to
significantly augment phagocytosis of
apoptotic cells by macrophages (Ren
and Savill, 1995). However, the precise
mechanisms involved in recognition
and elimination of apoptotic cells in
catagen HFs remain unclear.
Hair follicle cell populations that
survive catagen. Survival of distinct
HF cell populations during catagen is
essential for maintenance of HF regen-
eration in subsequent hair cycles. These
cell populations include fibroblasts of
the dermal papilla and connective tis-
sue sheath, KC and melanocyte stem
cells and some of their daughter (tran-
sient amplifying) cells (Ito et al., 2004;
Nishimura et al., 2005; Sharov et al.,
2005). Dermal papilla fibroblasts show
potent resistance to apoptosis associ-
ated with high levels of Bcl-2 and lack
of death receptors during the entire hair
cycle in mice and humans (Lindner et
al., 1997; Matsuo et al., 1998; Soma
et al., 1998). However, they may be
susceptible to apoptosis under certain
experimental conditions (application
of trypsin or staurosporine) or geneti-
cally altered conditions (hairless mice)
(Ferraris et al., 1997; Seiberg et al.,
1997; Panteleyev et al., 1999). Although
the majority of KC stem cells survive
during catagen, some bulge cells under-
go apoptosis (Ito et al., 2004). However,
the mechanisms and purpose of this
process remain to be further clarified.
Perspectives: Modulation of Apoptosis in
the Hair Follicle as a Tool for Correction
of Hair Growth Abnormalities
Defects in the regulation of apoptosis
contribute to many diseases, including
hair growth disorders (hirsutism and
various hair loss conditions; Paus and
Cotsarelis, 1999). Recently, new drugs
for modulating apoptosis (for example,
TNF--inhibiting antibodies, Bcl-2/Bcl-
x
L
antisense oligonucleotides, synthetic
p53, and caspase inhibitors) have been
identified, and some of them have been
successfully tested in clinical practice
(Komarov et al., 1999; Reed, 2002;
Kiechle and Zhang, 2002). It was dem-
onstrated that HFs in a number of hair
loss conditions (androgenetic alopecia,
chemotherapy-induced hair loss) show
alterations in apoptosis (Sawaya et al.,
2002; Botchkarev et al., 2000). Perhaps
apoptosis inhibitors can be used to
reduce apoptosis activity in the HFs
affected by distinct forms of alopecia.
Pharmacological suppression of TGF-
2 by plant extract promotes human
HF growth in vitro and delays in vivo
catagen development in mice (Tsuji et
al., 2003); this suggests that a suppres-
sor of TGF- might also be effective in
preventing male pattern baldness.
As an alternative to pharmacologi-
cal apoptosis manipulation, physi-
cal methods that are widely used for
management of unwanted hair growth
(laser- and light-induced hair removal)
are also capable of inducing catagen
and apoptosis in the HF (Figure 2f)
(Shander et al., 1999). These methods
are based on the selective absorption of
light by the HF chromophores, includ-
ing melanocytes. It remains to be deter-
mined whether these methods may be
combined with pharmacological apop-
tosis modulation for the correction of
hair growth abnormalities. Progress in
these areas of research will result in
the development of novel strategies for
therapeutic intervention in the deregu-
lated apoptotic process seen in the dis-
orders of hair growth.
C O N FLI C T O F I N T ER ES T
The authors state no conflict of interest.
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