Platelet a-Granules

P . Harrison, E . Martin Cramer

S UMMA R Y . Platelets contain a vast number of biologically active molecules within cytoplasmic
granules which are classified according to their respective distinct ultrastructures, densities and
content. The a-granule is a unique secretory organelle in that it exhibits further compartmentalization
and acquires its protein content via two distinct mechanisms : (1) biosynthesis predominantly at the
megakaryocyte (MK) level (with some vestigial platelet synthesis) (e .g . platelet factor 4) and
(2) endocytosis and pinocytosis at both the MK and circulating platelet levels (e .g . fibrinogen (Fg) and
IgG) . The currently known list of a-granular proteins continues to enlarge and includes many adhesive
proteins (e.g . Fg, von Willebrand factor (vWf) and thrombospondin (TSP)), plasma proteins (e .g . IgG
and albumin), cellular mitogens (e .g. platelet derived growth factor and TGFP), coagulation factors
(e.g. factor V) and protease inhibitors (e.g. a 2 -macroglobulin and a 2-antiplasmin). More recently the
inner lining of the m-granule unit membrane has been demonstrated to contain a number of
physiologically important receptors including glycoprotein llb/llla (a1Ib13 3 ) and P-selectin . The a-
granules originate from small precursor granules which can be observed budding from the trans-Colgi
network within the platelet precursor cell, the MK . During MK maturation the a-granules become
very prominent and are ultimately packaged into platelets during thrombopoiesis. The at-granular
contents are destined for release during platelet activation at sites of vessel wall injury and thus play
an important role in haemostasis, inflammation, ultimate wound repair and in the pathogenesis of
atherosclerosis .

Blood platelets are anucleate cytoplasmic fragments
derived from bone marrow megakaryocytes (MKs)
and play a fundamental role in haemostasis and
thrombosis . Figure l demonstrates the overall ultras-
tructure of a normal platelet section via transmission
electron microscopy and shows the presence of many
granules, relatively few mitochondria and two promi-
nent membrane structures : the surface connected
canalicular system (SCCS) which consists of a series
of channels extending from the plasma membrane
throughout the platelet cytoplasm, and the dense
tubular system (DTS) . For a complete review of
P. Harrison, PhD, Coagulation Research, Rayne Institute. St.
Thomas' Hospital, London, SE1 7EH, UK, E . Martin Cramer,
MD, INSEAM U-348, Hopital Lariboisiere, Paris, France .
Correspondence to : P. Harrison, PhD
platelet ultrastructure readers are referred to the
articles by White ." During MK maturation, and to
a limited extent in circulating platelets, protein
biosynthesis, trans-Golgi sorting via the regulated
pathway and degrading/non-degrading endocytosis/-
pinocytosis results in the formation and packaging
of distinct populations of storage granules . The
granules become very prominent in the mature MK
cytoplasm, and are ultimately distributed within
platelets during the complex processes of thrombopo-
iesis . As platelets retain only a limited biosynthetic
capacity' the organelles are all synthesized at the
MK level and are thus unlikely to be resynthesized
after release from circulating platelets .
Early studies on platelet granules were predomi-
nantly via biochemical analysis of releasates, lysates 4

Blood Reviews (1993) 7, 52-62

6 ", 1993 Lonpnen Group UK Ltd

Fig . 1 Section of a resting platelet displaying a typical discoid
shape, a circumferential band of microtubules (mt), the surface
connected canalicular system (sccs) throughout the entire
platelet cytoplasm, the dense tubular system (dts) (which is a
remnant from the endoplasmic reticulum), numerous granules of
which the most prominent are the x-granules (A) with their
characteristic nucleoid, the dense bodies (db) and various
organelles e .g . mitochondria (m) and numerous glycogen
granules .
and subcellular fractions` and demonstrated that
platelets contain a variety of biologically active
molecules within distinct organelle populations . How-
ever, the application of immunocytochemistry clearly
showed the presence of four types of granules within
the cytoplasm : 6
1 . a-granules which contain a wide variety of coagu-
lation/adhesive proteins, growth factors and protease
inhibitors involved in both primary and secondary
haemostatic mechanisms .
2 . Dense bodies or 6-granules which store serotonin,
adenine and guanine nucleotides . divalent cations
and inorganic phosphate .' Serotonin is unique in
that is freely exchangeable with extracellular pools
via an active transport mechanism .
3 . Primary lysosomes s or X-granules which contain
a number of hydrolytic enzymes including acid
hydrolases, cathepsin and heparitinase . Although
they have been assumed to be exclusively secretory
in nature there is some recent evidence to suggest
that they function in a similar fashion to those found
in other cells .'
4 . Peroxisomes containing catalase . 10
Upon platelet stimulation at sites of vessel wall
injury many of the granule contents are exocytosed
to enhance local haemostasis by promoting inter-
actions with other platelets, leukocytes, plasma
proteins and the vessel wall . However, it is apparent
that the contents of different granules are secreted at
different rates and not always released in parallel
under the same conditions of platelet activation . a-
granular secretion can occur independently upon
BLOOD REVIEWS 53
stimulation of platelets by weak agonists, whereas
lysosomal secretion requires a stronger stimulus
In this review we will focus on the morphology,
content, origin, fate and function of the a-granule .
a-Granule Formation
The a-granules are formed during MK maturation .
Early studies suggested that they arise from the trans-
Golgi complex, and more recent evidence suggest
that small immature granules increase in size to form
mature granules ." Immature MKs appear to contain
only a few mature a-granules and a large number of
small a-granular-like precursors 13 (Fig . 2) . Immunol-
ocalization of synthesized a-granular proteins e .g .
von Willebrand factor (vWf) or thrombospondin
(TSP) within immature bone marrow derived MK
reveal that they are associated with the Golgi
apparatus cisternae, associated vesicles and small
granules." However, during MK maturation they
enlarge becoming very prominent" and are ulti-
mately distributed during thrombopoiesis to give
between approximately 50 to 80 a-granules per
individual formed platelet, possibly accounting for
the heterogeneity in the densities of circulating
platelets .", " Further differences in observed platelet
densities may also result from selective a-granular
release in vivo under certain conditions . Although
the mature MK a-granules appear identical to those
within platelets, there is now cumulative evidence to
suggest that the platelet a-granules and even lyso-
somes can continue to acquire some of their content
either by vestigial de novo protein biosynthesis' or
the endocytic/pinocytic pathway (see section on the
origin of a-granular proteins) .'
Morphology
Mature a-granules are spherical, oval and sometimes
elongated structures with diameters ranging from 200
to 500 nm each enclosed by a unit membrane . 16
Unlike other platelet granules, they exhibit much
greater heterogeneity in ultrastructure and individual
staining patterns ." In many transmission electron
microscope sections they appear to be subdivided
into three main compartments based upon apparent
electron density and the distribution of various
immunolabelled proteins within the granule (Fig . 3) . 16
The electron-dense nucleoid is the most characteristic
region and has been demonstrated to co-localize with
a number of constituents including (3-thromboglobu-
lin ((3TG) (Fig . 3 .5), PF-4 and proteoglycans ." The
area adjacent to the nucleoid (Star) is not as electron
dense and is often associated with labelling for
fibrinogen (Fg) (Fig . 3 .4), TSP and albumin . The
peripheral electronluscent zone contains a small
number of tubules (up to 6) of approximately 20 nm
in diameter (Fig . 3 .1 and 3 .2), which correlate with
vWf immunostaining (Fig . 3 .3)," are completely
absent in severe vWD platelets'sae and reduced in
5 4 PLATELET a-GRANULES
Fig . 2 Section of a maturing megakaryocyte : a-granules are present in the cytoplasm (A) and their precursors (g) are observed
budding off the trans-Golgi (Go) cisternae with their nucleoids already formed . The demarcation membrane system (dms) is scattered
throughout the cytoplasm .
number in variant forms of vWD . 20 Furthermore, as
similar structures are observed in the vWf storage
organelles (Weibel-Palade bodies) of the vascular
endothelium" it is hypothesized that these tubules
represent the highly multimeric intracellular form of
vWf particularly as normal porcine platelets contain
many more of such tubules and ultra-high MW vWf
than human platelets. 10,2' The inner face of the unit
membrane of the granule also contains a distinct
intracellular pool (Fig . 3 .6) of various membrane
associated receptors (e.g . Gp IIb/IIIa and P-selectin) .
Content
The list of known a-granular molecules continues to
enlarge and includes a multifunctional array of
adhesive proteins, coagulation factors, cellular mitog-
ens, protease inhibitors and proteoglycans . 23 The a-
granular proteins can be broadly subdivided into two
distinct classes : (1) proteins specific or predominantly
synthesized within the MK/platelet lineage e .g .
3TG24,25 and platelet Factor 4 (PF-4); 25,26 (2) pro-
teins identical to those synthesized at other sites
which include a wide range of adhesive proteins,
plasma proteins and growth factors . This list includes
albumin, 27 factor V, 25 vWf, 29 von Willebrand Anti-
gen-II (vWf:AgII), 30 TSP, 31 histidine rich glyco-
protein (HRGP), 32 fibronectin (Fn)," Fg, 34
Vitronectin (Vn), 35 high molecular weight kininogen
(HMWK), 36 IgG, 57 IgA, IgM, 38 platelet-derived
growth factor (PDGF), 39 transforming growth factor-
p (TGF(3), 40 Cl inhibitor," two proteins from the
alternative complement pathway,42 endothelial cell
growth factor, 43 Protease Nexin-2 (PN-II) (Amyloid
beta-protein precursor-APP),' osteonectin, 45 plas-
minogen,46 plasminogen activator inhibitor-I (PAT-
1)," platelet derived collagenase inhibitor," protein
S, 49 a2 -antitrypsin, 50 a2-macroglobulin, 50 a2-anti-
plasmin 5l and multimerin (P155) . Many other
known platelet proteins may also be ultimately
localized within the a-granules e .g . Vascular per-
meability factor (VPF), tissue factor pathway inhibi-
tor (TFPI)53 and interleukin-l0 . 54 Furthermore, the
unit membrane of the granule has been shown to
contain a number of different receptor molecules
including glycoprotein IIb/IIIa (allb(3 3 ), ss P-Selectin
(GMP-140, PADGEM or CD-62) 55 and more
recently, GMP-33, 57 a 24kd GTP-binding protein"
and CD-36 (Gp TV) ." Recently, the adhesive protein
osteonectin has also been shown to localize on the
inner face of the a-granule in both platelets and
MKs.45 Unlike the plasma membrane proteins, Gp
IIb/IIIa and CD-36, P-Selectin and osteonectin are
only present within the a-granular membranes of
resting platelets and require platelet activation to
translocate to the plasma membrane . Thus specific
antibodies raised against internal platelet membrane-
associated proteins (including those associated with
dense body and lysosomal membranes) offer excellent
tools for direct analysis of platelet activation
status.60-67 Furthermore, the release of non-
membrane associated platelet-specific a-granular pro-
teins (e .g. PF4 and (3TG) into the plasma offers a
direct sensitive indicator of platelet secretion and can
also be used in monitoring various clinical
disorders ."`
Origin of a-granule Proteins
Prior to 1985, it was assumed that all a-granular
proteins were derived from MK biosynthesis . Early
studies on the origin and nature of a-granular
proteins provided no evidence for exchange between
plasma and platelet pools, as many proteins were

Fig . 3 Compartmentalization of the a -granule . Four distinct
zones are apparent : (1) an electronluscent zone opposite the dark
nucleoid (N) where up to 6 tubular structures are present
(arrows) ; (2) by transverse section the structures shown in (1)
are clearly identified as tubules (arrows) ; (3) immunolabelling for
vWf :Ag associated with the electronluscent zone ; (4) an
intermediary zone (arrows) with immunolabelling for Fg ; (5) the
dark nucleoid constituted of proteoglycans, PF 4 and
immunolabelled with anti-f,TG (arrows) ; (6) the limiting
membrane which contains several receptors including P-Selectin
and Go IIb/IIIa (arrow) which is also present in the plasma
membrane (pm) . Reproduced from the British Journal of
Haematology, 1990, Vol 74 (2), pp . 126 with kind permission of
the publishers, Blackwell Scientific Publications Limited .
found to be biosynthesized within MKs . 71 " 72 How-
ever, the discovery of a-granular IgG and albumin
suggested that an endocytic pathway was possible
within MKs and/or platelets 17,71 In 1987, Handa-
gama et al provided the first evidence for such a
pathway using the tracer protein horseradish peroxi-
dase 14 thus precipitating a number of experiments
where exogenous Fg, albumin and IgG were all
demonstrated to be rapidly endocytosed by MKs
both in vivo' S and in vitro 76 and ultimately expressed
within circulating platelet m-granules . 75,77 Treatment
of an a fibrinogenemic patient with Fg replacement
therapy also provided evidence for both time depen-
dent MK and direct platelet uptake of Fg . 7 B The
BLOOD REVIEWS 55
concentration of platelet a-granular proteins indicates
that Fg, fn and HMWK comprise a category of
ligands which are potentially packaged via a receptor-
mediated endocytic mechanism whereas IgG, IgM,
IgA and albumin are probably packaged via a fluid
phase endocytic process,'' particularly as their plate-
let concentrations correlate with their respective
plasma levels ." The latter process probably occurs
in conjunction with the receptor-mediated mechanism
as a small amount of ambient fluid is trapped when
membrane invagination and pinching occur during
vesicle formation . The accumulating evidence for the
endocytic pathway suggested that several a-granular
proteins may have dual origins and led to the
application of sensitive modern molecular biological
techniques to study expression of Fg, IgG and
albumin specific mRNA within purified MK S . 71 11
The majority of these latest studies suggest that Fg,
IgG and albumin are exclusively derived from the
plasma pool and are not synthesized within MKs
and platelets . Thus, the a-granular proteins can be
further subdivided into several classes based upon
their origins 73 (See Table 1) : (1) platelet specific
proteins derived from exclusive or predominant MK
rough-endoplasmic reticulum synthesis (e .g . PF4 and
RTG) and targeted to the regulated storage vesicles
in the trans-golgi network ; (2) MK synthesized
proteins which are also synthesized by other cell
types (e .g . vWF and Factor V) but packaged in the
same manner as above and (3) non-MK synthesized
proteins which are derived from the circulating
plasma pool (e .g . Fg and IgG) . Studies within
maturing MKs suggest that the synthesized proteins
are expressed very early in immature cells and can
be localized within the trans-Golgi network and
immature granules . In contrast Fg is expressed
comparatively late in MK maturation and not
detectable within the Golgi cisternae and associated
vesicles, thus supporting its external origin .' } Again
Fg expression in cultured MKs is totally dependent
upon an external source of protein' 3 .76 and there is
cumulating evidence for a novel role for Gp fib/111a
in mediating Fg endocytosis . s2-s5 Although there is
evidence to suggest that circulating platelets can also
acquire Fg directly,' 5 -' 8 the total contribution of
both MKs and platelets remains to be fully clarified .
As Gp l lb/I Ila is the receptor for Fg on platelets but
requires platelet activation before binding its
ligand(s), 86 it is conceivable that most of the a-
granular Fg content may be acquired at the MK
level . Nevertheless, there is plenty of evidence demon-
strating that Gp IIb/IHa is a dynamic receptor within
resting platelets cycling to and from intracellular
membranes i .e . within the SCCS and a-granules . e
In 1990, Wencel-Drake proposed the existence of two
models of Gp IIb,/IIIa/membrane turnover 87 :
(1) plasma membrane Gp IIb/IIIa is cleared by
lateral diffusion into the SCCS and (2) Gp IIb/IIIa
is internalized via endocytic vesicles which cycle to
and from the granules and the plasma mem-
brane,/SCCS . Although coated vesicles have pre-
56 PLATELET a-GRANULES
Table I Classification of alpha granular proteins
Proposed mechanism
of acquisition Protein Site of synthesis
MK/platelet vWF MK, endothelium
synthesis factor V MK, Liver
TSP MK, endothelium, etc
RTG MK
PF4 MK
Fluid-phase IgG B lymphocytes
endocytosis IgA B lymphocytes
Albumin Liver
Receptor-mediated Fn Endothelium, etc
endocytosis HMWK Endothelium
F9 Liver
Reproduced with kind permission of W . B . Saunders Co ., from George 7 . N . 1990. Blood 76 : 859-870 .
viously been demonstrated within platelets," it has
only recently been demonstrated that ligand/gold
complexes can be internalized within such structures
and ultimately fuse with a-granules .e 9,9 ° Further-
more, there is recent evidence for two pathways of
endocytosis 9 : ( 1) a coated vesicle non-degrading
pathway targeted to the a-granule and (2) a clathrin
independent degrading pathway targeted to the
lysosomes. There is at present no evidence for the
recycling of any of the other membrane associated
receptors .
a-Granule Fate
Studies into the mechanism(s) of platelet a-granule
release have led to the proposal of a number of
different secretory pathways . Several reports have
indicated that a-granules can fuse with the SCCS
and that the contents are extruded through small
channels of the plasma membrane ." , " Immunocyto-
chemical labelling of a-granule proteins demonstrates
that platelet activation with thrombin results in the
redistribution of a-granules and the SCCS into the
centre of the cell surrounded by the contracted
peripheral ring of microtubules . The granules begin
to fuse with the SCCS to form large intracytoplasmic
vacuoles which ultimately fuse with the plasma
membrane (Fig . 4) . Labelling with antibodies demon-
strates that the intracellular vacuole inner membranes
are Gp Iib/IIIa positive. Although the SCCS may
provide a mechanism for by-passing the classical
exocytic process (Fig . 4, inset) 93 there is some
evidence for direct fusion of a-granules with the
plasma membrane in response to C5b-9 stimulation .'
Furthermore, bovine platelets which contain little or
no SCCS can actively secrete alpha-granular proteins,
providing further evidence for the direct secretory
pathway ."
Function of a-granular Proteins
The multi-functional diversity of a-granular proteins
are required for promotion of effective local haemo-
stasis, inflammation and ultimate wound repair .
Upon sufficient strength of agonist stimulation, or-
Platelet concentration Plasma concentration Ratio :
(mg/ml) (mg/ml) platelet/plasma
0 .50 0 .01 50
0 .45 0 .007 64
6 .25 1 .3 x 10 - ° 48,000
4 .80 1 .8 x 10 -5 260,000
3 .30 1 .2x 10 -5 280,000
0 .52 11 .3 0.05
0 .05 2 .2 0.02
2 .53 42 .8 0 .06
0 .19 0.30 0.63
0 .06 0.08 0.75
7 .30 3 .0 2.43
granular release results in the exocytosis of proteins
at the site of injury to initially recruit other platelets,
leukocytes and plasma proteins to the vicinity .
Although some a-granular proteins are released into
the extracellular medium, many proteins remain
bound to the platelet plasma membrane after associ-
ation and exocytosis via a-granular Gp IIb/IlIa 96 or
re-association with plasma membrane Gp IIb/IIIa,
Gp lb or other components .9',98 Studies on the
functional relevance of many a-granular proteins are
often hampered by their relatively low concentrations
in the platelet and by the co-existence of extracellular
pools of some proteins . In this review we will
concentrate on the function of the major components
of the a-granular pool, as one can speculate that
many of the minor proteins may not have any
particular important role because of their very low
concentration . Three proteins critical for normal
haemostasis (vWF, Fg and factor V) appear at
relatively higher concentrations within the platelet a-
granular pool than in the plasma and thus may be
critical for normal haemostasis . Fg comprises 8% of
total platelet protein, is derived from the plasma
pool, and is absent or reduced in the platelets from
patients with Glanzmann's thrombasthenia (Gp
IIb/IIIa deficiency) 99 and congenital afibrinogene-
mia . 10 ° The role of a-granular Fg in haemostasis is
unknown particularly as afibrinogenemic platelets
aggregate normally in the presence of exogenous
added Fg . However, studies with afibrinogenemic
patients infused therapeutically with Fg have shown
that acquired a-granular Fg circulates longer in the
blood than does the infused plasma pool . 78J 1 °o
Sustained corrected bleeding times suggest that the
release of platelet intracellular Fg at sites of injury
may be important ."' In support of this observation,
the a-granular pool of Fg is as efficient as plasma Fg
in supporting platelet aggregation ."' a-granular Fg
is often surface expressed, particularly at granule
discharge sites possible via a Gp IIb/IIIa mediated
translocation mechanism . 102 One can observe the
redistribution of a-granular-matrix Fg to the inside
of the granule membranes during the initial stages of
thrombin activation, apparently associating with Gp
IIb/IIIa .ro3

Fig . 4 After activation with appropriate stimuli e .g . a-thrombin, the platelet becomes spheroid with pseudopods (p), contracts and
releases the contents of the cytoplasmic granules into the dilated channels of the SCCS (D) . Dense knots of microfilaments remain in
the centre of the platelet (mF) . Inset : Part of a activated platelet section immunolabelled for Fg : a-granules (A) release their content into
the dilated SCCS (D) .
Unlike Fg, MKs synthesize vWf 104 and thus
analysis of vWf in platelet lysates provides support
of the clinical diagnosis and classification of vWD, 1o5
particularly as platelet vWf may or may not exhibit
the same multimeric/functional abnormality as
plasma vWf. In normal subjects, intraplatelet vWf is
comprised of higher MW multimers,' O6 which are
more effective in haemostasis and represents 25% of
the total circulating pool . Many studies have demon-
strated the importance of platelet vWf in haemostasis .
'The classic study by Bowie et al correlated platelet
vWf with bleeding time in pigs where normal bone
marrow was transplanted into severe vWD pigs ."'
For a comprehensive review readers are referred to
the paper of Gralnick et ." al Clinical observations
in vWD and the porcine model mentioned above
suggest that providing platelet vWf is normal, plasma
levels of vWf can be as low at 5 to 10% of normal
without affecting normal haemostasis . Platelet vWf
can be expressed on the plasma membrane possibly
via Gp IIb/IIIa translocation or by rebinding to
either Gp lb or Gp IIb/Illa 97 and probably functions
in promoting both spreading on exposed subendothel-
ium and inter-platelet bridging .
Platelets contain around 18 to 25% of the total
circulating pool of factor V within the a-granules
which is synthesized by MKs .'2 Although platelet
activation by various agonists results in factor V
secretion, only thrombin stimulation results in the
conversion of factor V to the activated form Va . ws
It has been proposed that platelet factor V may
function as the factor Xa binding site on platelets
resulting in the ultimate generation of the prothrom-
binase complex . Evidence for this hypothesis is
provided by studies in factor V deficient platelets
where the number of Xa binding sites are
decreased ."'
BLOOD REVIEWS 57
TSP is another multifunctional glycoprotein
involved in cell-cell and cell-matrix interactions and
a major constituent of the a-granules . TSP forms
multi-macromolecular complexes with other adhesive
proteins e .g . Fg and Gp IIb/Illa, Fn and
CD36 . 10-173 The TSP-Fg-Gp IIb/Illa complexes are
probably responsible for irreversible platelet aggre-
gation ."' Furthermore, TSP can also mediate the
interaction of activated platelets with monocytes via
bridging between CD-36 on both cells . Both a-
granular Fn and Vn are again expressed on the
activated platelet surface and may participate in
inflammatory and repair processes at the site of tissue
damage .' t4
The expression of a-granular membrane P-selectin
on the platelet surface has been demonstrated to
promote platelet-leukocyte interactions . 115 118 P-se-
lectin is a member of the cell surface glycoproteins,
the 'selectins' which all use carbohydate structures to
promote adhesion of neutrophils and monocytes .
P-selectin may provide an important mechanism for
restriction of both inflammatory and haemostatic
processes to the site of vessel injury ."'
The recent finding that protease nexin-II is localized
within a-granules 119 and secreted during platelet
activation is interesting as the protein is the precursor
molecule of amyloid (3-protein which is deposited
within plaques in patients with Alzheimer's disease ."'
However, the protein exhibits potent protease inhibi-
tor and growth factor activity"' and probably plays
an important role in wound repair . In Alzheimer's
disease it is possible that subtle alterations occur in
both platelet release and processing of APP leading
to the accumulation of amyloid (3-protein within the
central nervous system vasculature .
(3-TG is specific to the MK lineage and is
multifunctional, binding to and inhibiting prostacy-
58 PLATELET a-GRANULES

clin production of endothelial cells' •• and is a potent

fibroblast chemotactic agent . 113 PF4 exhibits potent
antiheparin or glycosaminoglycan binding (e .g .
heparan sulfate) and is chemotactic for neutrophils
and monocytes . PDGF stimulates proliferation of
arterial smooth muscle cells and fibroblasts 124 and
may eventually result in the development of athero-
sclerosis . For a complete review of their functions
and properties the reader is referred to the articles
by Deuel et al 125 and Levine,' 26 In particular a-
granular proteins have been demonstrated to modu-
late the uptake of oxidized low density lipoprotein
(LDL) into macrophages 127,128 e.g . PDGF stimulates
and (3TG inhibits macrophage uptake . These inter-
actions may have implications in foam cell formation
particularly as platelets are often in close proximity
to macrophages in the arterial wall .
Platelets also contain a large pool of TGFP1' 2s
which exhibits a wide range of biological activities
including inhibition of MK growth and endom-
itosis .' 3 o Other platelet proteins within serum and
platelet lysates e .g . PF4"' also inhibit MK growth
so it is feasible that a-granular proteins may function
in regulating thrombopoiesis via feedback mechan-
isms to the bone marrow .

a-Granule Abnormalities
The important role(s) of the platelet granules in
haemostasis have been further indicated by the study
of patients with various congenital platelet disorders,
characterized by the absence of one or more of the
granule types . A combination of biochemical and
electron microscopic analysis of platelets from
patients with storage pool disorders (SPD) reveals
three distinct groups of patients 132 all generally
characterized by different degrees of impaired platelet
function and defective aggregation responses : (1) S-
SPD with decreased dense granule content ; (2) aS-
SPD with a marked deficiency of dense granules with
a parallel variable deficiency of a-granules 132 and
(3) a-SPD or Gray platelet syndrome (GPS) with
loss of a-granules only In the latter group the
platelets not only appear gray after Romanovsky
staining but exhibit decreased aggregation responses
to ADP, collagen and thrombin in vitro and thus
offer an excellent model for studying the role of a-
granules in haemostasis . Biochemical analysis of GPS
platelets reveals a severe deficiency but not complete
absence of synthesized a-granular proteins and near
normal levels of pinocytosed proteins e .g . albumin
and IgG 134. 13 e all which immunolocalize within a
population of small granules (<0 .1 µm in diameter)
(Fig . SA) within platelets and MKs . 1.." ' These
granules appear similar to those granules observed
budding from the trans-golgi complex in immature
MKs and may represent a-granular precursors par-
ticularly as their membranes are often positive for
P-selectin and Gp llb/Illa . P-selectin labelling is also
observed in abnormal vacuolar structures but not in
Fig . 5 A : platelet from a patient with Gray platelet syndrome
(GPS) lacking normal a-granules but demonstrating the
presence of small granules (g), empty vacuolar structures (v),
normal SCCS, mitochondria (m) and Golgi complex (Go) .
B : Gray platelet immunolabelled for P-selectin, demonstrating
that GPS platelets contain a normal amount of a-granular
membrane within small granules (g) and vacuoles (v) . The
SCCS is apparently not labelled . m=mitochondria .
the SCCS (Fig. 5B) . The presence of these granules
and associated proteins would suggest that normal
a-granular protein synthesis and endocytosis still
occur and that the defect is caused by a defect in
targeting of synthesized proteins to the granules,
resulting in their early release . The resulting high
concentrations of a-granular growth factors within
the bone marrow stroma are probably involved in
the pathogenesis of myelofibrosis .' 3s
Conclusions
The a-granule represents a unique secretory organelle
in that it is highly compartmentalized and acquires
its protein content via a combination of biosynthetic
and endocytic mechanisms predominantly within the
MK and to some extent within the circulating
platelet . The importance of the granule in haemostasis
is emphasized by studies on patients with GPS and
in clinical conditions where a-granules are prema-
turely released . Measurement of released membrane
bound or soluble a-granular proteins provides a
number of excellent tools for classification of various
primary haemostatic disorders and monitoring plate-
let activation status both in vivo and in vitro . The a-

granules are a significant intracellular storage pool
of a battery of proteins vital to normal haemostasis,
inflammation and wound healing . Future studies into
the cell biology of a-granules and their proteins will
eventually increase our understanding of the targeting
signals and receptors involved in packaging and
release mechanisms of this unique organelle .
Acknowledgements
The authors are grateful to Dr R . H . Saundry for his helpful
comments, Daniele Tenza for technical help with electron
microscopy and Association pour In Recherche sur le Cancer for
financial support .
References
l . White J G. Clawson C C. Overview article: biostructure of
blood platelets . Ultrastructural Pathology 1980 ; 1 : 533-558 .
2 . White J G. Anatomy and structural organization of the
platelet . In : Colman R W et al eds) Haemostasis and
Thrombosis (2nd Ed) . J B Lippincott Company,
Philadelphia, pp. 537-554 .
3 . Kieffer N, Guichard J . Farcet J P, Vainchenker W, Breton
Gorius J . Biosynthesis of major platelet proteins in human
blood platelets . European Journal of Biochemistry 1987 : 164:
189-195 .
4 . Bezkorovainy A, Rafelson M E. Characterisation of some
proteins from normal human platelets . Journal of
Laboratory and Clinical Medicine 1964 ; 64: 212-225 .
5 . Broekman M J, Westmoreland N P. Cohen P . An improved
method for isolating alpha granules and mitochondria from
human platelets . Journal of Cell Biology 1974; 60 : 507-519 .
6 . Bentfeld M E, Bainton D F . Cytochemical localization of
Ivsosomal enzymes in rat megakaryocytes and platelets .
Journal of Clinical Investigation 1975 ; 56 : 1635-1649 .
7 . Meyers K M, Holmsen H, Seachord C L. Comparative study
of platelet dense granule constituents . American Journal of
Physiology 1982: 243 : R454-8461 .
8 . Bentfeld-Barker M E, Bainton D F . Identification of primary
lysosomes in human megakaryocytes and platelets . Blood
1982 ;59 :472--481 .
9 . Behnke O . Degrading and non-degrading pathways in fluid-
phase (non-adsorptive) endocytosis in human blood platelets .
Journal of Submicroscopy Cytology Pathology 1992 ; 24 :
169-178 .
10 . Breton-Gorius J, Guichard J . Ultrastructural localization of
peroxidase activity in human platelets and megakaryocytes .
American Journal of Pathology 1972 ; 66 : 277 286 .
11, Kaplan K L, Broekman M J . ChemoBA . Lesznik G R,
Drillings M. Platelet alpha-granule proteins: studies on
release and subcellular localization . Blood 1979 ; 53 : 604-618 .
12 . Holmsen H, Kaplan K L, Dangelmaier C A . Differential
requirements for platelet responses : A simultaneous study of
dense, alpha-granule and acid hydrolase secretion .
Biochemical Journal 1982 ; 208 : 9-18 .
13 . Cramer F M . Harrison P, Savidge G F et a] Uncoordinated
expression of alpha-granule proteins in human
megakaryocytes . Progress in Clinical and Biological
Research 1990 :356 :131-142 .
l4 . Corash L, Costa I L, Shafer B . Donlon 1 A, Murphy D .
Heterogeneity of human whole blood platelet
subpopulations .III. Density-dependent differences in
subcellular constituents . Blood 1984; 64 : 185-193 .
15 . Vicic W J . Weiss H 1 . Evidence that platelet alpha-granules
are a major determinant of platelet density : studies in storage
pool deficiency. Thrombosis and Haemostasis 1983 ; 50 :
878-880 .
16 . Harrison P. Savidge G F, Cramer E M . The origin and
physiological relevance of alpha-granule adhesive proteins .
British Journal of Haematology 1990 ; 74: 125-130 .
17 . Spicer S S, Green W B, Hardin J H . Ultrastructural
localisation of acid mucosaccharides and antimonale-
precipitable cation in human and rabbit platelets and
Rr OOD REVIEWS co
megakaryocytes . Journal of Histochemistry and
Cytochemistry 1969 ; 17 :781-792 .
18 . Cramer E M, Meyer D, le Menn R . Breton
Gorius J . Eccentric localization of von Willebrand factor in
an internal structure of platelet alpha-granule resembling
that of Weibel-Palade bodies . Blood 1985: 66 :710-713 .
19 . Cramer E M, Caen I P, Drouet L, Breton Gorius J . Absence
of tubular structures and immunolabeling for von
Willebrand factor in the platelet alpha-granules from porcine
von Willebrand disease . Blood 1986 ; 68 : 774-778 .
20 . Wilbourn B R. Harrison P, Lawrie A S, Savariau E, Savidge
G F, Martin Cramer E . Porcine platelets contain an
increased quantity of ultra-high molecular weight von
Willebrand factor . British Journal of Haematology 1993 (In
press).
21 . Wagner D D, Olmsted J B . Marder V J . lmmunolocalization
of von Willebrand protein in Weibel-Palade bodies of human
endothelial cells . Journal of Cell Biology 1982; 95 : 355-360.
22 . Cramer E M, Breton Gorius J . Beesley J F, Martin J F.
Ultrastructural demonstration of tubular inclusions
coinciding with von Willebrand factor in pig
megakarvocytes . Blood 1988 ; 71 : 1533-1538 .
23 . Schick B P . Proteoglycans in megakarvocyte development .
Progress in Clinical and Biological Research 1990; 356 :
93-104.
24 . Sander H J, Slot J W, Bouma B N . Bolhuis P A, Pepper D S .
Sixma 11 . Immunocytochemical localization of fibrinogen,
platelet factor 4. and beta thromboglobulin in thin frozen
sections of human blood platelets . Journal of Clinical
Investigation 1983 ;72 :1277-1287 .
25 . McLaren K M, Pepper D S . Immunological localisation of
beta-thromboglobulin and platelet factor 4 in human
megakaryocytes and platelets . Journal of Clinical Pathology
1982 ;35 :1227-1231 .
26 . Pham T D, Kaplan K L, Butler V P, Jr . Immunoelectron
microscopic localization of platelet factor 4 and fibrinogen in
the granules of human platelets . Journal of Histochemistry
and Cytochemistry 1983 ; 31 : 905-910 .
27 . Sixma J J, van den Berg A, Schiphorst M, Geuze H J,
Mcdonagh J. Immunocytochcmical localization of albumin
and factor XIII in thin cryo sections of human blood
platelets . Thrombosis and Haemostasis 1984; 51 : 388-391 .
28 . Chesney C M, Pifer D, Colman R W . Subcellular localization
and secretion of factor V from human platelets . Proceedings
of the National Academy of Science USA 1981 ; 78 :
5180-5184 .
29 . Slot J W, Bouma B N, Montgomery R, Zimmerman T S .
Platelet factor VIII-related antigen : Immunofluorescent
localization . Thrombosis Research 1978 : 13 : 871-881 .
30 . Scott J P . Montgomery R . Platelet von Willebrand antigen
H : active release by aggregating agents and a marker of
platelet release reaction in vivo . Blood 1981 ; 58 : 1075-1080 .
31 . Jaffe E, Leung L L K, Nachman R L, Levin R, Mosher D F .
Thrombospondin is the endogenous lectin of human
platelets . Nature 1982 ; 295 : 246-248 .
32 . Leung L L K, Harpel PC ., Nachman R L, Rabellino E M .
Histidine rich glycoprotein is present in human platelets and
is cleared during thrombin stimulation . Blood 1983 ; 62 :
1016-1021 .
33 . Zucker M B, Mosesson M W, Broekman M J . Kaplan K L.
Release of platelet frbronectin (cold-insoluble globulin) from
alpha granules induced by thrombin or collagen : lack of
requirement for plasma fibronectin in ADP-induced platelet
aggregation . Blood 1979 ; 54 : 8-12 .
34 . Broekman M J, Handin R I, Cohen P . Distribution of
fibrinogen, and platelet factors 4 and XIII in subcellular
fractions of human platelets . British Journal of Haematology
1975 ;31 :51-55 .
35 . Preissner K T, Holzhuter S . Justus C, Muller Berghaus G .
Identification of and partial characterization of platelet
vitronectin : evidence for complex formation with platelet-
derived plasminogen activator inhibitor-l . Blood 1989 ; 74 :
1989-1996 .
36 . Schmaier A H, Zuckerberg A, Silverman C et al . High
molecular weight kininogen, a secreted platelet protein .
Journal of Clinical Investigation 1983 ; 71 : 1477-1489 .
37 . George J N, Saucerman S, Levine S P, Knieriem L K ..
Balaton D F . Immunoglobulin G is a platelet alphaa granule-
secreted protein . Journal of Clinical Investigation 76 :
2020 2025 .
AB Pr ATFI FT n-r:R ANI II cc
38 . George J N, Saucerman S . Platelet IgG, IgA, IgM, and
albumin : correlation of platelet and plasma concentrations in
normal subjects and in patients with ITP or dysproteinemia .
Blood 72: 362-365.
39 . Heldin C H, Westermark B, Wasteson C . Platelet derived
growth factor: purification and partial characterisation .
Proceedings of the National Acadamy of Sciences USA 1979 ;
76:3722-3726.
40 . Assoian R K, Komorija A, Meyers C A, Miller D M, Spent
M B . Transforming growth factor-beta in human platelets .
Identification of a major storage site, purification and
characterisation . Journal of Biological Chemistry 1983 ; 258:
7155-7160.
41 . Schmaier A H, Smith P M, Colman R W . Platelet Cl-
inhibitor. A secreted alpha-granule protein . Journal of
Clinical Investigation 1985 ; 75 : 242-250 .
42 . Kenney P M, Davis A E . Association of alternative
complement pathway components with human blood
platelets : Secretion and localization of Factor D and I H
globulin . Clinical Immunology Immunopathology 1981 ; 21 :
351-363 .
43 . King G L, Buchwald S. Characterization and partial
purification of an endothelial cell growth factor from human
platelets . Journal of Clinical Investigation 1984 ; 73 : 392-396 .
44 . Van Nostrand W E, Schmaier A H, Farrow J S, Cines D B,
Cunningham D . Protease nexin-2/amyloid beta-protein
precursor in blood is a platelet-specific protein . Biochemical
Biophysical Research Communications 1991 ; 175 : 15-21 .
45 . Breton Gorius J, Clezardin P, Guichard J et al . Localization
of platelet osteonectin at the internal face of the alpha-
granule membranes in platelets and megakaryocytes . Blood
1992; 79 :936-941 .
46. Holt J C, Niewiarowski S. Secretion of Plasminogen by
washed human platelets (abstr) . Circulation 1980 ; 62: 342
(suppl 3).
47 . Booth N A, Simpson A J, Croll A, Bennett B, MacGregor
I R . Plasminogen activator inhibitor (PAI-1) in plasma and
platelets . British Journal of Haematology 1988; 70 : 327-333 .
48 . Cooper T W, Eisen A Z, Stricklin G P, Welgus H G . Platelet-
derived collagenase inhibitor: characterization and
subeellular localization . Proceedings of the National
Academy of Sciences of the USA 1985 ; 82 : 2779-2783 .
49. Schwarz H P, Heeb M J, Wencel Drake J D, Griffin J H .
Identification and quantitation of protein S in human
platelets . Blood 1985; 66 : 1452-1455 .
50. Nachman R L, Harpel P C . Platelet alpha2-macroglobulin
and alpha (-antitrypsin . Journal of Biological Chemistry
1976;251 :4512-4521 .
51 . Gogstad G O, Stormorken H, Solum N O. Platelet alpha
2-antiplasmin is located in the platelet alpha-granules .
Thrombosis Research 1983 ; 31 : 387-390 .
52. Hayward C P M, Smith J W, Horsewood P, Warkentin T E,
Kelton J G. p-155, a multimerie platelet protein that is
expressed on activated platelets . Journal of Biological
Chemistry 1991 ; 266 : 7114-7120 .
53 . Novotny W F, Girard T J, Miletich J P, Broze G J . Platelets
secrete a coagulation inhibitor functionally and antigenically
related to the lipoprotein associated coagulation inhibitor .
Blood 1988 ; 78 : 2020-2025.
54. Hawrylowicz C M, Sanotoro S A, Platt F M, Unanue E R .
Activated Platelets express IL-I activity . Journal of
Immunology 1989; 143 : 4015-4018 .
55 . Cramer E M, Savidge G F, Vainchenker W et al . Alpha-
granule pool of glycoprotein lib-Illa in normal and
pathologic platelets and megakaryocytes . Blood 1990 ; 75 :
1220-1227 .
56 . McEver R P . Properties of GMP-140, an inducible granule
membrane protein of platelets and endothelium . Blood Cells
1990;16 :73-80 .
57 . Metzelaar M J, Hcijnen H F, Sixma J J, Nieuwenhuis H K .
Identification of a 33-Kd protein associated with the alpha-
granule membrane (GMP-33) that is expressed on the surface
of activated platelets . Blood 1992 ; 79 : 372-379 .
58 . van der Meulen J, Bhullar R P, Chancellor Maddison K A .
Association of a 24-kDa GTP-binding protein, Gn24, with
human platelet alpha-granule membranes . FEBS Letters
1991 ;291 :122-126 .
59 . Berger G, Tenza D, Caen J P, Martin Cramer E .
Glycoprotein IV is found on the platelet alpha granule
membrane . Blood 1992 ; 80:132a (abstract) .
60 . Israels S J, Gerrard J M, Jacques Y V, McNicol A, Cham B,
Nishibori NI, Bainton D F . Platelet dense granule
membranes contain both granulophysin and P-selectin
(GMP-140) . Blood 1992 ; 80: 143-152.
61 . Gerrard J M, Lind D, Sims P J et al . Identification of a
platelet dense granule membrane protein that is deficient in a
patient with the Hermansky-Pudlack syndrome . Blood 1991 ;
'77 : 101-12 .
62 . Metzelaar M J, Clevers H C . Lysosomal membrane
glycoproteins in platelets . Thrombosis and Haemostasis
1992 ;68 :378-382 .
63 . Febbraio M, Silverstein R L . Identification and
characterization of LAMP-I as an activation-dependent
platelet surface glycoprotein . Journal of Biological Chemistry
1990 ;265 :18531-18537 .
64 . Metzelaar M J, Sixma J J, Nieuwenhuis H K . Detection of
platelet activation using activation specific monoclonal
antibodies . Blood Cells 1990 ; 16 : 85 93 .
65 . Corash L . Measurement of platelet activation by
fluorescence-activated flow cytometry . Blood Cells 1990 ; 16 :
97-106 .
66 . Abrams C S, Ellison N, Budzynski A Z, Shattil S J . Direct
detection of activated platelets and platelet-derived
microparticles in humans . Blood 1990; 75 : 128- 138 .
67 . Shattil S J, Cunningham M, Hoxie J A . Detection of
activated platelets in whole blood using activation-dependent
monoclonal antibodies and flow cytometry . Blood 1987 ; 70:
307-315 .
68 . Kaplan K L, Owen J . Plasma levels of platelet secretory
proteins. Critical Reviews in Oncology/Haematology 1986 ; 5 :
235-255 .
69 . Dawes J, Clemetson K J, Gogstad G O et al . A
radioimmunoassay for thrombospondin, used in a
comparative study of thrombospondin, beta-
thromboglobulin and platelet factor 4 in healthy volunteers .
Thrombosis Research 1983 ; 29 : 569-581 .
70 . Nichols A B, Owen J, Kaplan K L, Sciacca R R, Cannon P l,
Nossel H L . Fibrinopeptide A, platelet factor 4, and beta-
thromboglobulin levels in coronary heart disease . Blood
1982 ;60 :650-654 .
71 . Leven R M, Schick P K, Budzinski A M . Fibrinogen
biosynthesis in isolated Guinea Pig megakaryocytes . Blood
1985 ;65 :501-504 .
72 . Chin H C, Schick P K, Colman R W . Biosynthesis of factor
V in isolated guinea pig megakaryocytes . Journal of Clinical
Investigation 1985 ; 75: 339-346.
73 . George J N . Platelet immunoglobulin G : its significance for
the evaluation of thrombocytopenia and for understanding
the origin of alpha-granule proteins . Blood 1990 ; 76 :
859-870 .
74 . Handagama P J, George J N, Shuman M A, McEver R P,
Bainton D F . Incorporation of a circulating protein into
megakaryocyte and platelet granules . Proceedings of the
National Academy of Sciences in the USA 1987 ; 84 :
861-865 .
75 . Handagama P J . Shuman M A, Bainton D F . Incorporation
of intravenously injected albumin, IgG and fibrinogen in
guinea pig megakaryocyte granules . Journal of Clinical
Investigation 1989 ; 84: 73-82.
76 . Cramer E M, Debili N, Martin J F et al . Uncoordinated
expression of fibrinogen compared with thrombospondin and
von Willebrand factor in maturing human megakaryocytes .
Blood 1989; 73 : 1123-1129 .
77 . Handagama P J, Shuman M A, Bainton D F . In vivo
defibrination results in markedly decreased amounts of
fibrinogen in rat megakaryocytes and platelets . American
Journal of Pathology 1990 ; 137 : 1393-1399 .
78 . Harrison P, Wilbourn B, Debili N et al . Uptake of plasma
fibrinogen into the alpha granules of human megakaryocytes
and platelets . Journal of Clinical Investigation 1989 ; 84 :
1320-1324.
79 . Lange W, Luig A, Dolken G, Mertelsmann R, Kanz L .
Fibrinogen gamma-chain mRNA is not detected in human
megakaryocytes . Blood 1991 ; 78 : 20-25 .
80 . Handagama P, Rappolee D A, Werh Z, Levin J, Bainton
D F . Platelet alpha-granule fibrinogen, albumin, and
immunoglobulin G are not synthesized by rat and mouse
megakaryocytes . Journal of Clinical Investigation 1990 ; 86:
1364-1368 .
81 . Louache F, Debili N, Cramer E M, Breton-Gorius J,

Vainchenker W . Fibrinogen is not synthesized by human
megakaryocytes . Blood 1991 ; 77 : 311-316 .
82 . Suzuki M . Kawakatsu T, Nagata H et al . Effects of injected
antibody against the platelet glycoprotein IIb/IIIa complex
on monkey platelet fibrinogen . Thrombosis and Haemostasis
1992 ;67 :578-581 .
83 . Harrison P . Platelet a-granular fibrinogen . Platelets 1992 : 3 :
I l0 .
84. Harrison P, Wilhourn B R, Cramer E M et al . The Influence
of therapeutic blocking of Gp IIb/IIIa on platelet a-granular
fibrinogen . British Journal of Haematology 1992 ; 82 :
721-728 .
85 . Coller B S . Seligsohn U, West S M . Scudder L E, Norton
K J . Platelet fibrinogen and vitronectin in Glanzmann
thrombasthenia : evidence consistent with specific roles for
glycoprotein IIb/Jifa and a„/1, integrins in platelet protein
trafficking . Blood 1991 ;78 :2603-2610.
86 . Kieffer N, Phillips D R . Platelet membrane glycoproteins :
function in cellular interactions . Annual Review in Cell
Biology 1990; 6 :329-357 .
87 . Wencel-Drake J D . Plasma membrane Gp IIb/IIIa : evidence
for a cycling receptor pool . American Journal of Pathology
1988;136:61-70.
88 . Morganstern E. Coated membranes in blood platelets .
European Journal of Cell Biology 1982 ; 26 : 315-3 I8 .
89 . Morgenstern E, Ruf A, Patscheke H . Transport of anti-
glycoprotein IIb/Illa-antibodies into the alpha granules of
unstimulated human blood platelets . Thrombosis and
laemostasis 1992: 67: 121-125 .
90 . Behnke O. Coated Pits and vesicles transfer plasma
components to platelet granules . Thrombosis and
Haemostasis 1989 ; 62 : 718-722 .
91 . White J G, Krumwiede M . Further studies of the secretory
pathway in thrombin-stimulated human platelets . Blood
1987 ; 69 :1196 1203 .
92 . Escolar G, White J G . The platelet open canalicular system : a
final common pathway . Blood Cells 17 : 467-485 .
93 . White 3 G, Eseolar G . The blood platelet open canalicular
system : a two-way street. European Journal of Cell Biology
1991 ;56 :233-24i .
94 . Sims P J . Faioni E M . Wiedmer T, Shattil S J . Complement
proteins C5h-9 cause release of membrane vesicles from the
platelet surface that are enriched in the membrane receptor
for coagulation factor Va and express prothrombinase
activity . Journal of Biological Chemistry 1988 ; 283 :
18205-18212 .
95 . Zucker-Franklin D, Benson K A, Myers K M . Absence of a
surface-connected canalicular system in bovine platelets .
Blood 1985;65 :241-244.
96 . Legrand C, Dubernard V, Nurdcn A T . Studies on the
mechanism of expression of secreted fibrinogen on the
surface of activated human platelets . Blood 1989; 73 :
1226 1234.
97 . Parker R I, Gralnick H R . Identification of platelet
glycoprotein Ilb/IIIa as the major binding site for released
platelet von Willebrand factor . Blood 1986 ; 68 : 732-736 .
98 . George J N, Onofre A R . Human platelet surface binding of
endogeneous secreted factor VIII-von Willebrand factor and
platelet factor 4 . Blood 1982 ; 59 : 194-197 .
99 . George J N, Caen J P, Nurden A T . Glanzmann
thrombasthenia : a spectrum of clinical disease . Blood 1990 ;
75 :1383-1395.
100 . Cattaneo M, Bettega D, Lombardi R, Lecchi A, Mannucci
P M . Sustained correction of the bleeding time in an
afibrinogenemic patient after infusion of fresh frozen plasma .
British Journal of Haematology 1992; 82 : 388-390.
101, Kaplan K L, Dauzier M J, Rose S . ADP and epinephrine
induced release of platelet fibrinogen . Blood 1981 ; 58 :
797-802 .
102 . Suzuki H, Kinlough-Rathbone R L. Packham M A,
Tanoue K, Yamasaki H, Mustard J F . Immunocytochemical
localisation of fibrinogen during thrombin-induced
aggregation of washed human platelets . Blood 1988; 71 :
1310-1320 .
103 . Suzuki H, Baba 1, Tanoue K, Yamazaki H . A possible role of
a-granule Gp IIb/IIIa as a carrier of intragranular fibrinogen
during release reaction . Thrombosis and Haemostasis 1991 ;
65 : 688 (abstract).
104 . Sporn L A, Chavin S I, Marder V J, Marder V J, Wagner
D D . Biosynthesis of von Willebrand protein by human
BLOOD nrvrrwc 61
megakaryocytes . Journal of Clinical Investigation 1985; 76 :
1102-1106 .
105 . Gralnick H R, Williams S B, Mckeown L P . Magruder L,
Hausmann K, Vail M . Platelet von Willebrand factor . Mayo
Clinic Proceedings 1991 ; 66: 634-640 .
106 . Gralnick H R, Williams S B, McKeown L P, Krizek D M,
Shafer B C, Rick M E . Platelet von Willebrand factor :
comparison with plasma von Willebrand factor . Thrombosis
Research 1985; 38 : 623-633 .
107 . Bowie E J W, Solberg L A . Fass D N, Johnson C M,
Knutson G J, Stewart M L, Zoecklin L l . Transplantation of
normal bone marrow into a pig with severe von Willebrand's
disease. Journal of Clinical Investigation 1986 ; 78 : 26-30 .
108 . Tracy P B, Mann K G . A model for the assembly of
coagulation factor complexes on cell surfaces : Prothrotnbin
activation on platelets. In : Phillips D R, Shuman M A (eds)
Biochemistry of Platelets . Academic Press Inc (London) Ltd .
1986 : pp . 295-318 .
109 . Miletich J P, Majerus D W . Majerus P W . Patients with
congenital factor V deficiency have decreased factor Xa
binding sites on their platelets . Journal of Clinical
Investigation 1978; 62 : 824-831 .
110 . Leung L L K, Nachman R L. Complex formation of platelet
thrombospondin with fibrinogen. Journal of Clinical
Investigation 1982 : 70: 542-549 .
111 . Lahav J, Schwartz M A, Hynes R O. Analysis of platelet
adhesion with a radioactive cross linking reagent : interaction
of thrombospondin with fibronectin and collagen . Cell 1982 ;
31 :253-362 .
112 . Leung L L K . Role of thrombospondin in platelet
aggregation . Journal of Clinical Investigation 1984 : 74 :
1764-1772 .
113 . Aiken M L, Ginsburg M A, Plow E F . Identification of a
new class of inducible receptors on platelets,
thrombospondin interacts with platelets via a Gp IIb/IIIa
independent mechanism. Journal of Clinical Investigation
1986 ; 78 : 1713 1716 .
114 . Parker C J . Stone 0 L, White V F, Bernshaw
N J . Vitronectin is associated with blood platelets . British
Journal of Haematology 71 : 245-252 .
115 . Stenberg P E . McEver R P, Shuman M A . Jacques Y V,
Balaton D F. A platelet alpha-granule membrane protein
(GMP-140) is expressed on the plasma membrane after
activation . Journal of Cell Biology 1985 ; 101 : 880-886 .
116. Hamburger S A. McEver R P . GMP-140 mediates adhesion
of stimulated platelets to neutrophils . Blood 1990 ; 75 :
550-554 .
117 . Larsen E, Celi A, Gilbert G E . Furie B C, Erban 1 K .
Bonfanti R, Wagner D D, Furie B . PADGEM protein : a
receptor that mediates the interaction of activated platelets
with neutrophils and monocytes . Cell 1989; 59: 305 312 .
118 . Palabrica T, Lobb R, Furie B C, Aronovitz M, Benjamin C,
Hsu Y M, Sajer S A, Furie B . Leukocyte accumulation
promoting fibrin deposition is mediated in vivo by P-selectin
on adherent platelets . Nature 1992 ; 359 : 848-851 .
119 . Van Nostrand W E, Schmaier A H, Farrow J S, Cunningham
D D. Protease Nexin H (amyloid beta-protein precursor) : a
platelet alpha-granule protein . Science 1990 ;248 : 745-748 .
120 . Van Nostrand W E, Schmaier A H, Farrow J S, Cunningham
D D. Platelet protease nexin-2/amyloid beta-protein
precursor . Possible pathologic and physiologic functions .
Annals of the New York Academy of Science 1991 ; 640 :
140-144 .
121 . Van Nostrand W E, Wagner S L, Farrow J S . Cunningham
D D. Immunopurification and protease inhibitory properties
of protease nexin-2/amyloid beta-protein precursor . Journal
of Biological Chemistry 1990 ; 265 : 9591-9594 .
122 . Hope W, Martin T J, Chesterman C N, Morgan F J . Human
(3-thromboglobulin inhibits PGI 1 production and binds to a
specific site in bovine aortic endothelial cells . Nature 1979 ;
282 :210-212 .
123 . Senior R M, Griffin G L, Huang J S, Walz D A, Dcuel T F .
Chemotactie activity of platelet alpha granule proteins for
fibroblasts . Journal of Cell Biology 1983 ; 96 : 382-385 .
124. Ross R, Vogcl A . The platelet derived growth factor . Celt
1978 ;14:203-210 .
125 . Deuel T F, Huang S S, Huang J S . Platelet derived growth
factor: purification, characterization and role in normal and
abnormal cell growth . In : Phillips D R, Shuman M A (eds)
F7 PT ATPI PT n-f7R AHII II PS
Biochemistry of Platelets, Academic Press Inc (London) Ltd.
1986 :pp .347-375 .
126 . Levine S P . Secreted Platelet proteins as markers for
pathological disorders . In : Phillips D R, Shaman M A (eds)
Biochemistry of Platelets . Academic Press Inc (London) Ltd .
1986 ; pp . 377-415 .
127 . Fuhrman B, Brook G J, Aviram M . Activated platelets
secrete a protein-like factor that stimulates oxidized-LDL
receptor activity in macrophages . Journal of Lipid Research
1991 ;32 :1113-1123 .
128 . Fuhrman B, Brook G J, Aviram M . Proteins derived from
platelet alpha granules modulate the uptake of oxidized low
density lipoprotein by macrophages . Acta Biochimica
Biophysica 1992:1127: 15-21 .
129 . Fava R A, Casey T T, Wilcox J, Pelton R W, Moses H L,
Nanney L B . Synthesis of transforming growth factor 0 by
megakaryocytes and its localization to megakaryocyte and
platelet x-granules . Blood 1990; 76 :1946-??,
130 . Kuter D J, Gminski D M, Rosenberg R D . Transforming
growth factor (3 inhibits megakaryocyte growth and
endomitosis . Blood 1990; 79 : 619-626,
131 . Gerwitz A M, Calabretta B, Rucinski B, Niewiarowski S, Xu
W Y . Inhibition of human megakaryocytopoiesis in vitro by
platelet factor 4 (PF4) and a synthetic COOH-terminal PF4
peptide. Journal of Clinical Investigation 1989 ; 83 :
1477-1486 .
132 . Lages B, Shattil S J, Bainton D F, Weiss H J . Decreased
content and surface expression of alpha-granule membrane
protein GMP-140 in one of two types of platelet alpha delta
storage pool deficiency . Journal of Clinical Investigations
1991 ;87 :919-929 .
133 . Levy Toledano S, Caen J P, Breton Gorius J et al Gray
platelet syndrome : alpha-granule deficiency . Its influence on
platelet function . Journal of Laboratory and Clinical
Medicine 1981 ; 98 : 831-848.
134 . Gerrard I M . Phillips D R, Rao G H et al Biochemical
studies of two patients with the gray platelet syndrome .
Selective deficiency of platelet alpha granules . Journal of
Clinical Investigation 1980 ; 66 : 102-109 .
135 . White J G . Ultrastructural studies of the gray platelet
syndrome. American Journal of Pathology 1979 ; 95 :
445-462 .
136 . Rosa J P, George J N, Bainton D F, Nurden A T, Caen J P,
McEver R P. Gray platelet syndrome . Demonstration of
alpha granule membranes that can fuse with the cell surface .
Journal of Clinical Investigation 1987 ; 80 : 1138-1146 .
137 . Cramer E M, Vainchenker W, Vinci G, Guichard J, Breton
Gorius J . Gray platelet syndrome : immunoelectron
microscopic localization of fibrinogen and von Willebrand
factor in platelets and megakaryocytes. Blood 1985 ; 66 :
1309-1316 .
138 . Caen J P, Deschamps J F, Bodevin E, Bryckaert M C,
Dupuy E . Wasteson A . Megakaryocytes and myelofrbrosis in
gray platelet syndrome . Nouvelle Revue Francaise
Hematologe 1987 ; 29 : 109-114 .