Cold-induced activation of human platelets is associated with the passage of the platelets through their thermotropic phase transition. Anti-freeze glycoproteins (AFGPs) from the blood of certain Arctic and Antarctic fishes are capable of stabilizing human platelets during chilling and storage below the phase transition (4 C) for up to 21 days in vitro. Recent studies suggest that these AFGPs stabilize platelets in part, by preventing lateral phase separation of platelet membrane phospholipids. Such storage conditions would most certainly increase the ease of blood banking this valuable resource, however, prior to human clinical trials, it is essential to determine whether AFGPs can stabilize platelets from several potential animal models. The specific goals of the current project are to: (1) determine if passage the thermotropic phase transition correlates with cold-activation in a variety of animal model systems and to elucidate underlying events during storage, (2) to provide evidence concerning the mechanism by which AFGPs inhibit these events, including possible inhibition of microvesiculation and lateral phase separation, (3) determine the AFGP- platelet recovery and survival times in our animal model systems in vivo. Several techniques will be employed to accomplish these goals. Fourier-transform infrared spectroscopy will be used to monitor the membrane phospholipid phase transitions and lateral phase separations. Fluorescence spectroscopy will be employed for measuring vesiculation and lateral phase separation. Fluorescence activated cell scanning will measure the expression of glycoproteins Ib, IIb-IIIa and P-selectin on the plasma membrane, as well as to determine platelet recovery and survival times in vivo. In addition we have developed a unique shapemeter to quantify the level of shape change, activation and agonist responsiveness of these platelets during and after cold-storage.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL061204-04
Application #
6390074
Study Section
Special Emphasis Panel (ZHL1-CSR-N (S1))
Program Officer
Barbosa, Luiz H
Project Start
1998-09-30
Project End
2002-08-31
Budget Start
2001-09-01
Budget End
2002-08-31
Support Year
4
Fiscal Year
2001
Total Cost
$196,802
Indirect Cost
Name
University of California Davis
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
047120084
City
Davis
State
CA
Country
United States
Zip Code
95618
Field, Cara L; Tablin, Fern (2012) Response of Northern Elephant Seal platelets to pressure and temperature changes: a comparison with human platelets. Comp Biochem Physiol A Mol Integr Physiol 162:289-95
Bali, Rachna; Savino, Laura; Ramirez, Diego A et al. (2009) Macroscopic domain formation during cooling in the platelet plasma membrane: an issue of low cholesterol content. Biochim Biophys Acta 1788:1229-37
Tang, Minke; Wolkers, Willem F; Crowe, John H et al. (2006) Freeze-dried rehydrated human blood platelets regulate intracellular pH. Transfusion 46:1029-37
Oldenhof, Harriette; Wolkers, Willem F; Bowman, John L et al. (2006) Freezing and desiccation tolerance in the moss Physcomitrella patens: an in situ Fourier transform infrared spectroscopic study. Biochim Biophys Acta 1760:1226-34
Krishnan, V V; Lau, Edmond Y; Tsvetkova, Nelly M et al. (2005) Characterization of the restricted rotation of the dimethyl groups in chemically N-terminal 13C-labeled antifreeze glycoproteins: a temperature-dependent study in water to ice through the supercooled state. J Chem Phys 123:044702
He, Xiaoming; Wolkers, Willem F; Crowe, John H et al. (2004) In situ thermal denaturation of proteins in dunning AT-1 prostate cancer cells: implication for hyperthermic cell injury. Ann Biomed Eng 32:1384-98
Leidy, Chad; Gousset, Karine; Ricker, Josette et al. (2004) Lipid phase behavior and stabilization of domains in membranes of platelets. Cell Biochem Biophys 40:123-48
Wolkers, Willem F; Oliver, Ann E; Tablin, Fern et al. (2004) A Fourier-transform infrared spectroscopy study of sugar glasses. Carbohydr Res 339:1077-85
Gousset, Karine; Tsvetkova, Nelly M; Crowe, John H et al. (2004) Important role of raft aggregation in the signaling events of cold-induced platelet activation. Biochim Biophys Acta 1660:7-15
Wolkers, Willem F; Oldenhof, Harriette; Tablin, Fern et al. (2004) Preservation of dried liposomes in the presence of sugar and phosphate. Biochim Biophys Acta 1661:125-34

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