HEMATOLOGY 2 LECTURE: PLATELET STRUCTURE (Jan. 31-PM) J. BENEDICTO – M. DRAPER-N.

INOCENTES

PLATELET STRUCTURE - The plasma membrane has a phospholipid bilayer and a semipermeable
membrane. It is the barrier of the external and internal environment
A. Chromomere/Granulomere -Fluid Mosaic Model
▪ Located centrally and is granular * Ngaa Mosaic- kay may proteins and cholesterol
▪ Wright’s Stain: PLATELET =PURPLE/VIOLET. Why? Tungod sa * Ngaa Fluid? Ano ang ga favor sa movement/fluidity sang membrane?
granules ya. That’s why gina tawag man siya nga Chromomere kay siya CHOLESTEROL. That’s why if gamay ang cholesterol sa membrane maging
man ang rason nga ga take up sang stain ang platelet rigid siya.
B. Hyalomere - hydrophilic- head (outside);hydrophobic-tail (inside)
▪ Surrounds the chromomere and is nongranular or clear to light blue - glycoprotein- serves as receptor.
- what would happen if may ma bind sa receptor? Platelet Activation. What
▪ Sa TEM, makita ang microtubules. Ang microtubules naga palibot, will happen next? (hindi ko di sure if ano next kay super active si ate mace sa
circumferencial. Siya ga maintain sang shape ni platelet. class, iya voice mabatian ko haha)
▪ Light stained- alpha granules. Darkly stained- dense. - Adhesion > Activation > Change in Shape
▪ Open canalicular system(OCS)- channel nga gina gwaan sang substances - basta may nagtapik sa receptor ni platelet maacdtivate gd na dayon
▪ Neutral Phospholipids (outer leaflet)
ZONE AND COMPONENT  Phosphatidylcholine
A. Peripheral Zone -  Spingomyelin
B. Membrane System  Charge- Neutral
C. Structural or Sol-Gel Zone ▪ Anionic or Polar Phospholipids (inner leaflet)
D. Organelle Zone - maintains platelet structure  Negatively charged phospholipid kay anionic
 Phosphatidylethanolamine
A. PERIPHERAL ZONE  Phosphatidylinositol
▪ Contains plasma membrane − Purpose in the plasma membrane: Needed for
▪ Cholesterol allows fluidity of membrane production of arachidonic acid
− Prostaglandin and thromboxane - converted from
 PLASMA MEMBRANE (phospholipid, submembrane area, PF3) arachidonic acid
 Phosphatidylserine
− Flips to the outer surface serve as charged phospholipid
− Cause of the negativity of the activated platelet -
− Carries the negative charge
− Activated platelet- negatively charge
− Ma transfer sa outer leaflet. That’s why negatively charge
ang platelet after activation
 GLYCOCALYX (Phospholipid, submembrane area, PF3)
▪ 20-30 nm
▪ Exterior coat with glycoprotein receptor sites - outside of the plasma
membrane
▪ Location of receptors
▪ Absorbs albumin, fibrinogen, and other plasma proteins
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 ▪ Platelet does not the synthesize fibrinogen gina absorb ya lang
▪ Sponge like structure ni platelet kay naga pang absorb siya especially HI kamo kay Ate Mace. Kay ga Hi naman siya!! Haha

plasma proteins
B. MEMBRANE SYSTEM
▪ Endocytosis - method of absorption
▪ fibrinogen is an endocytosed (?) protein
 OPEN CANALICULAR MEMBRANE SYSTEM
Other name: Surface-connected canalicular system
 GLYCOPROTEIN RECEPTOR
Surface connected- connected to the plasma membrane
Glycoprotein Platelet Membrane Receptors That Participate In Adhesion And the ▪ Route for endocytosis and for secretion of alpha-granule contents
Initiation of Aggregation by Binding Specific Ligands ▪ Two way- pwede pasulod, pwede man paguwa
Electropho  DENSE TUBULAR SYSTEM
Cluster
resis Current
Ligand Design Comments ▪ One way only- secretion only (paguwa lang)
Nomenclat Nomenclature
ation ▪ Sequesters Ca 2+
ure
GP Ia/IIa Integrin: α2β1 Collagen CD29, Avidity is upregulated via ▪ Bears series of enzymes for platelet activation
CD49b “inside out” activation ▪ Phospholipase A2 , Cyclooxygenase, Thromboxane synthetase
Integrin: αvβ1 Vitronectin − Enzymes for platelet activation
Integrin: α5β1 Laminin CD29, − Needed eicosanoid synthetic pathway
CD49e − Controls platelet activation
Integrin: α6β1 Fibronectin CD29,
CD49f C. STRUCTURAL OR SOL-GEL ZONE
GP IV CAM of the Collagen Key collagen receptor,
immunoglobulin triggers activation, release
gene family  CIRCUMFERENTIAL MICROTUBULES
of agonists that the avidity
▪ Kung isa lang tubulin, pero if damo(bundles of tubulin) or long:
of integrins α2β1 and αIIbβ3
magiging microtubules na ang tawag diyan
GP Ib/IX/V CAM of the VWF and CD42a, GP Ib/IX/V is a 2:2:2:1
leucine-rich thrombin CD42b complex of GP Ibα and Ibβ ,
▪ Cylindrical on cross section kay naga palibot siya sa plasma
repeat family bind GP Ibα ; CD42c, GP IX and GP V. There are membrane
thrombin CD42d 25,000 copies on the ▪ Parallel the plane of the outer surface
cleaves a site resting platelet membrane ▪ During platelet activation:
on GP V surface, 5% to 10% on the 1. Move inward to enable the expression of alpha-granule contents
alpha granule membrane, - It will contract and will eventually lead to change in shape thus
but few on the SCCS allowing movement
membrane. GP Ibα is the 2. Reassemble in long parallel bundles to provide rigidity
VWF-specific site. 50% of
Ibα/Ibβ is cleared from the
membrane on the
Given the situation ma activate si platelet or not?
activation. Benard-Soulier > Microtubules nag form sang bundle? ACTIVATE
syndrome mutation are (basta nag change ang shape may activation nga na tabo)
identified for all but GP C.
Bound to subsurface actin-  MICROFILAMENTS (Actin)
binding protein ▪ Anchors the plasma membrane Gp and proteoglycans
GP IIb/IIIa Integrin: αIIbβ3 Fibrinogen, CD41, GP IIb and IIIa are ▪ Anchors receptors on the plasma membrane
VWF CD61 distributed on the surface ▪ Present throughout the cytoplasm (20%-30% of platelet protein)
membrane, SCCS and alpha
1. Resting platelet(inactive platelet): globular and amorphous
granule membranes (30%).
2. Increased cytoplasmic calcium: filamentous and contractile
Heterodimer forms on
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  INTERMEDIATE FILAMENTS (Desmin and Vimentin) Dense Granule (Dense Body) Contents
▪ Ropelike polymers (8-12 nm) Small Molecule Comment
▪ Connect with actin and the tubules ADP Nonmetabolic, supports neighboring platelet
▪ Anchors between microtubules and microfilament. That’s why it is aggregation by binding to ADP receptors P2Y1, P2Y12
called INTERMEDIATE FILAMENTS kay sa tunga siya. ATP Function unknown, but ATP release is detectable
▪ Sa histopath ta lang daw eh get-to-know sila Desmin and Vimentin upon platelet activation
Serotonin Vasoconstrictor that binds endothelial cells and
Functions of Sol-Gel Zone: (in order)
platelet membranes
1. Platelet shape change
Ca2+ and Mg2+ Divalent cations support platelet activation and
2. Extension of pseudopods
coagulation
3. Secretion of granule contents
First, may contraction then eventually may change in shape (ga gamay ang
- Calcium and Magnesium eh remember gd nga positive charge sila
cytoplasm) causing pseudopods to be extended. If nag extend or nag laba na
Positively-charged calcium is needed for activation of coagulation factor.
ang pseudopods ma open ang OCS para may secretion of granule content.
Calcium mediator between negatively-charged platelet and coagulation factor.
D. ORGANELLE ZONE - ATP is a by product of platelet aggregation. If ATP level is high in patient
meaning may platelet activation thus releasing damo nga energy because
active
▪ Mitochondria - powerhouse of the cell (ari gd ni sa iya ang korona. Amo
gd ni permi role ya) - ADP (for platelet aggregation) recruits/triggers other platelets to go to the
site of injury
▪ Glycogen - storage for carbohydrate derivatives
- ADP helps express GP IIb/IIIa
▪ Granules - important structure
 Dense granules(darkly stained)- Storage non-protein mediators - ADP - major factor that helps in platelet aggregation
 Alpha-granules (lightly stained)- Storage of protein mediators
ALPHA-GRANULES
▪ Lysosomes and Peroxisomes - contains enzymes and lipids
Stain positive for the ff cytochemical stain:
 Arylsulfatase ▪ 50-80 per platelet (granulation)
 Beta-glucuronidase ▪ Stain medium gray in osmium-dye TEM
▪ Release of granule contents:
 Acid phosphatase - main enzyme
 Catalase  Fuse with the SCCS/OCS
 Contents flow to the nearby microenvironment
DENSE GRANULES VS ALPHA GRANULES
COAGULATION PROTEINS NONCOAGULATION
PROTEINS
DENSE GRANULES
Proteins Present in Platelet Cytoplasm and Alpha-Granules
▪ 2-7 per platelet (granulation) Endocytosed Fibronectin Albumin
▪ Appear later than alpha-granules Fibrinogen Immunoglobulins
▪ Stain black with osmium dye in TEM Megakaryocyte- Factor V -
▪ Release of granule contents: synthesized Thrombospodin -
 Migrate to the plasma membrane VWF -
 Release contents directly into the plasma Proteins Present in Alpha Granules but not in the cytoplasm
Megakaryocyte- Beta-thromboglobulin EGF
Synthesized HMWK Multimerin

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 PAI-1 PDC1 PRESENT IN ALPHA-GRANULES ONLY
Plasminogen PDGF
PF4 TGF-beta  Megakaryocyte synthesized only
Protein C Inhibitor VEGF/VPF  Beta-thromboglobulin
Platelet Membrane Bound Proteins ▪ Chemoattractant for neutrophils, fibroblasts
▪ Neutralizes heparin - thus may clotting kay inhibited si heparin
Restricted to alpha- P-selectin GMP33
▪ Procoagulant
granule membrane - Osteonectin
 Protein S, TFPI, Alpha1 Protease Inhibitor, C1 Esterase Inhibitor
In alpha-granule and GP IIb/IIIa Cap1 ▪ Inhibitors of coagulation
plasma membrane GP IV CD9 ▪ Platelet favors coagulation and at the same time it can inhibit
GP Ib/IX/V PECAM-1 coagulation due to the proteins nga ara sa iya
 Plasminogen
Diin halin ang mga proteins? ▪ Converted to plasmin (fibrinolysis)
- ENDOCYTOSED- from the plasma. Kung manotice mo puro plasma proteins  PAI-1
AGAIN WALA GINA PRODUCE NI PLATELET ANG FIBRINOGEN, GINA ABSORB ▪ Fibrinolytic inhibitor
YA LANG!!  Platelet Factor 4 (PF4)
- MEGAKARYOCYTE-SYNTHESIZED- ara mismo kay platelet kay halin siya sa ▪ Neutralizes heparin
mother ni platelet ang megakaryocyte. ▪ Suppresses megakarycytopoiesis
▪ Weak chemoattractant for neutrophils, fibroblasts
Receptors ang proteins sa membrane bound proteins :) ▪ Same function with beta-thromboglobulin
 PDGF, TGF, EGF, IGF
PRESENT IN PLATELET CYTOPLASM AND ALPHA-GRANULES ▪ Mga growth factors helps in wound healing and repair
▪ Promotes regrowth of smooth muscle cells (wound repair)
 Fibrinogen ▪ Mitogenic factors
▪ Platelet aggregation
▪ converted into fibrin SUMMARY: DENSE GRANULES vs ALPHA GRANULES
▪ Converted to fibrin to form the fibrin clot
▪ May coagulation factor involved GRANULE DENSE ALPHA
 Factor V, XI Number of Granules 2-7 per platelets 50-80 per platelets
▪ Fibrin formation Staining Darkly Stained Lightly Stained
 Von Willebrand factor
Release Mechanism Direct release to plasma Fuse with the OCS
▪ Indirect platelet adhesion (primary)
Visible color with Osmium Black Medium Gray
▪ (secondary) vWF is the carrier of factor VIII in plasma (maskin damo
Dye
ang Factor VIII but if wala ang vWF, hndi man siya gyapon ka
participate sa coagulation pathway. Kung sa love pa, one sided lang
ti kundi wala gid chance maglevel up ang relationship) PLATELET SECRETION
▪ vWF-VIII
▪ From the endothelial cell and alpha granule  Alpha-granules and lysosomes: SCCS
▪ Platelets, Fibrinogen and vWF - involved sa primary and secondary ▪ Coagulation proteins
 Thrombospondin, Fibronectin, Vitronectin (receptors ni sila)  Dense granules
▪ Adhesive glycoproteins ▪ Migrate to the plasma membrane
▪ Vasoconstrcictors
▪ Platelet agonists
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 PLATELET ACTIVATION PATHWAY - The last enzyme, Thromboxane Synthase will convert prostaglandin to
thromboxane.
G PROTEIN MECHANISM - Ano dayon ang purpose ni thromboxane? For Platelet activation.
- Mabalik naman sa thromboxane sa cycle.
- REPEAT PATHWAY NAMAN
- if sige sige ang cycle, madamo man ang thromboxane mas bongga ang
platelet activation. Ti kundi bongga. (Daw ikaw, sige sige pakatanga sa iya kundi
sige sige man ang activation sang luha mo )
- What if sa endothelial cell ang cycle? Instead of Thromboxane synthase
maging Prostacyclin synthase. End product, prostacyclin = Platelet inhibitor.

Endothelial cell > Prostacyclin synthase > Prostacyclin > Platelet inhibitor
Platelet > Thromboxane Synthase > Thromboxane > Platelet Activator

For Hemostasis:
- Sa platelet activation dapat may gastop man. And ang mastop sa iya is ang
prostacyclin from the endothelial cell (Dapat gid man abi kung tam’an kana ka
tanga sa sigesige na lang may ga tampa mn simo para magbugtaw kag maguntat.)
-If wala may ma bind sa receptor, inactive. If may ara then activated.
-G-proteins are activated by Ligand or agonist SECOND MESSENGER SYSTEM

EICOSANOID SYNTHESIS

- Thromboxane suppresses adenylate cyclase causing the decrease in cyclic

AMP
- Ang decrease in cyclic AMP is done para ang calcium sa DTS maguwa.
- Increase cytoplasmic calcium leads to platelet activation
- Pero ang calcium ara lang sa DTS, inactive ang platelet
- Starting sa receptor, may nagbind nga agonist (ADP, Thrombin, Collagen,
Epinephrine)
- Phosphatidylinositol converted to arachidonic acid with the help of the
enzyme Phospholipase A2..
- (DENSE TUBULAR SYSTEM) The arachidonic acid, with the aid of
Cycloxygenase(enzyme), will be converted to PgG2 . The same with the enzyme
Peroxidase ma convert man to PgH2

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