Intracellular function in rehydrated lyophilized platelets.
This study aimed to evaluate the effect of cross-linking and lyophilization on intracellular signalling processes in rehydrated, lyophilized (RL) platelets, which are under development as a platelet substitute for transfusion. Exposure of RL platelets to thrombin resulted in enhanced phosphorylation of several proteins, including 18 kDa and 42 kDa kinase substrates that were shown to be the substrates of myosin light chain and protein kinase C respectively. Cross-linking and lyophilization depleted the platelets of free cytoplasmic ADP and ATP, but had less effect on protein-bound nucleotides. The surface membrane of RL platelets was found to be permeable to poly dT probes less than approximately 3 kDa in size; larger nucleotide probes and proteins did not penetrate the surface membrane. Taken together, our results indicate that RL platelets retain some of the haemostatic stimulus-response functions of fresh platelets and are capable of feedback amplification in coagulation.
Fischer
T H
TH
Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 27599, USA. tfischer@med.unc.edu
Merricks
E P
EP
Russell
K E
KE
Raymer
R A
RA
White
G C
GC
Bode
A P
AP
Nichols
T C
TC
Read
M S
MS
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D001345:Autoradiography; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology*; D015153:Blotting, Western; D002462:Cell Membrane / Q000502:physiology; D003593:Cytoplasm / Q000201:enzymology; D005612:Freeze Drying; D006801:Humans; D018994:Myosin Light Chains; D015345:Oligonucleotide Probes / Q000378:metabolism; D010539:Permeability; D010766:Phosphorylation; D017713:Platelet Transfusion; D011493:Protein Kinase C; D015398:Signal Transduction; D013917:Thrombin / Q000494:pharmacology*
D018994:Myosin Light Chains; D015345:Oligonucleotide Probes; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011493:Protein Kinase C; D013917:Thrombin