TY - JOUR
T1 - Platelet storage under in vitro condition is associated with calcium-dependent apoptosis-like lesions and novel reorganization in platelet cytoskeleton
AU - Wadhawan, Vinita
AU - Karim, Zubair A.
AU - Mukhopadhyay, Saikat
AU - Gupta, Ramkrishna
AU - Dikshit, Madhu
AU - Dash, Debabrata
N1 - Funding Information:
This work was supported in part by grants received by D. Dash from the Council of Scientific and Industrial Research, Indian Council of Medical Research, R.D. Birla Smarak Kosh, and the Department of Biotechnology, Government of India. The donation from Alexander von Humboldt-Stiftung, Germany, is gratefully acknowledged. The electron microscopic study was carried out in the Department of Anatomy, AIIMS, New Delhi. We thank Dr. Peter Presek and Dr. James Powers for their generous gift of antibodies and reagents.
PY - 2004/2/15
Y1 - 2004/2/15
N2 - Platelets are cleared from circulation after a life span of 8-10 days. The molecular mechanisms underlying platelet senescence remain poorly characterized. Here we report that, progressive functional impairment in the platelets incubated in vitro in a plasma-free isotonic medium for up to 24h at 37°C is associated with release of cytochrome c from platelet mitochondria and cleavage of procaspase-9, but without evidence of caspase-3 activation. Concomitantly, there was proteolysis of survival proteins like focal adhesion kinase, Src, gelsolin, and specific cytoskeleton-associated peptides, in a manner regulated by extracellular calcium and calpain activity. Cytoskeleton played a critical role as evidenced from the association of these proteins and their degradation products, as well as procaspase-3 and the actin regulatory small GTPase, CDC42Hs, with the cytoskeleton of the stored platelets. The cytoskeletal enrichment with specific proteins was not associated with increase in the content of F-actin and was cytochalasin-resistant, thus signifying a novel mechanism of interaction of the translocating proteins with the pre-existing cytoskeleton. There was progressive exposure of phosphatidylserine on the outer leaflet of platelet membrane and specific electron microscopic changes suggestive of apoptotic lesions. Based on these observations we discuss the caspase-independent but calpain-mediated signaling events in the stored platelets resembling the features of apoptosis in the nucleated cells.
AB - Platelets are cleared from circulation after a life span of 8-10 days. The molecular mechanisms underlying platelet senescence remain poorly characterized. Here we report that, progressive functional impairment in the platelets incubated in vitro in a plasma-free isotonic medium for up to 24h at 37°C is associated with release of cytochrome c from platelet mitochondria and cleavage of procaspase-9, but without evidence of caspase-3 activation. Concomitantly, there was proteolysis of survival proteins like focal adhesion kinase, Src, gelsolin, and specific cytoskeleton-associated peptides, in a manner regulated by extracellular calcium and calpain activity. Cytoskeleton played a critical role as evidenced from the association of these proteins and their degradation products, as well as procaspase-3 and the actin regulatory small GTPase, CDC42Hs, with the cytoskeleton of the stored platelets. The cytoskeletal enrichment with specific proteins was not associated with increase in the content of F-actin and was cytochalasin-resistant, thus signifying a novel mechanism of interaction of the translocating proteins with the pre-existing cytoskeleton. There was progressive exposure of phosphatidylserine on the outer leaflet of platelet membrane and specific electron microscopic changes suggestive of apoptotic lesions. Based on these observations we discuss the caspase-independent but calpain-mediated signaling events in the stored platelets resembling the features of apoptosis in the nucleated cells.
KW - Apoptosis
KW - Calpain
KW - Caspases
KW - Cytochrome c
KW - Cytoskeleton
KW - Platelet storage
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U2 - 10.1016/j.abb.2003.12.024
DO - 10.1016/j.abb.2003.12.024
M3 - Article
C2 - 14759606
AN - SCOPUS:0842283383
SN - 0003-9861
VL - 422
SP - 183
EP - 190
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 2
ER -