Platelet Dynamics during Natural and Pharmacologically Induced Torpor and Forced Hypothermia

Hibernation is an energy-conserving behavior in winter characterized by two phases: torpor and arousal. During torpor, markedly reduced metabolic activity results in inactivity and decreased body temperature. Arousal periods intersperse the torpor bouts and feature increased metabolism and euthermic body temperature. Alterations in physiological parameters, such as suppression of hemostasis, are thought to allow hibernators to survive periods of torpor and arousal without organ injury. While the state of torpor is potentially procoagulant, due to low blood flow, increased viscosity, immobility, hypoxia, and low body temperature, organ injury due to thromboembolism is absent. To investigate platelet dynamics during hibernation, we measured platelet count and function during and after natural torpor, pharmacologically induced torpor and forced hypothermia. Splenectomies were performed to unravel potential storage sites of platelets during torpor. Here we show that decreasing body temperature drives thrombocytopenia during torpor in hamster with maintained functionality of circulating platelets. Interestingly, hamster platelets during torpor do not express P-selectin, but expression is induced by treatment with ADP. Platelet count rapidly restores during arousal and rewarming. Platelet dynamics in hibernation are not affected by splenectomy before or during torpor. Reversible thrombocytopenia was also induced by forced hypothermia in both hibernating (hamster) and non-hibernating (rat and mouse) species without changing platelet function. Pharmacological torpor induced by injection of 5′-AMP in mice did not induce thrombocytopenia, possibly because 5′-AMP inhibits platelet function. The rapidness of changes in the numbers of circulating platelets, as well as marginal changes in immature platelet fractions upon arousal, strongly suggest that storage-and-release underlies the reversible thrombocytopenia during natural torpor. Possibly, margination of platelets, dependent on intrinsic platelet functionality, governs clearance of circulating platelets during torpor.

[1]  W. Milsom,et al.  Regulation of cardiac rhythm in hibernating mammals. , 1999, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[2]  E. Lechler,et al.  Blood clotting defect in hibernating ground squirrels (Citellus tridecemlineatus). , 1963, The American journal of physiology.

[3]  I. Schwarzinger,et al.  The new hematology analyzer Sysmex XE-2100: performance evaluation of a novel white blood cell differential technology. , 2001, Archives of pathology & laboratory medicine.

[4]  T. Mayadas,et al.  The Clearance Mechanism of Chilled Blood Platelets , 2003, Cell.

[5]  Siegfried Krause,et al.  Monitoring the effects of platelet glycoprotein IIb/IIIa antagonists with a microtiter plate method for detection of platelet aggregation , 2001, Platelets.

[6]  Gerhard Heldmaier,et al.  Natural hypometabolism during hibernation and daily torpor in mammals , 2004, Respiratory Physiology & Neurobiology.

[7]  M. B. Zucker,et al.  Reversible alterations in platelet morphology produced by anticoagulants and by cold. , 1954, Blood.

[8]  John H. Zhang,et al.  Platelet–Leukocyte–Endothelial Cell Interactions after Middle Cerebral Artery Occlusion and Reperfusion , 2004, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[9]  I. Bakaltcheva,et al.  Effect of taurine on platelets and the plasma coagulation system , 2002, Platelets.

[10]  A. Tilles,et al.  The near-wall excess of platelet-sized particles in blood flow: its dependence on hematocrit and wall shear rate. , 1987, Microvascular research.

[11]  Hjalmar R. Bouma,et al.  Low body temperature governs the decline of circulating lymphocytes during hibernation through sphingosine-1-phosphate , 2011, Proceedings of the National Academy of Sciences.

[12]  Hannah V Carey,et al.  Hibernation: the immune system at rest? , 2010, Journal of leukocyte biology.

[13]  P. Bray,et al.  Platelets of Female Mice Are Intrinsically More Sensitive to Agonists Than Are Platelets of Males , 2004, Arteriosclerosis, thrombosis, and vascular biology.

[14]  Penick Gd,et al.  Thrombocytopenia of hibernation. Mechanism of induction and recovery. , 1973 .

[15]  G. Körtner,et al.  THE TEMPORAL ORGANIZATION OF DAILY TORPOR AND HIBERNATION: CIRCADIAN AND CIRCANNUAL RHYTHMS , 2000, Chronobiology international.

[16]  R. W. Bullard,et al.  Estimated regional blood flow by rubidium 86 distribution during arousal from hibernation. , 1962, The American journal of physiology.

[17]  T. Mayadas,et al.  Hypoxia-induced exocytosis of endothelial cell Weibel-Palade bodies. A mechanism for rapid neutrophil recruitment after cardiac preservation. , 1996, The Journal of clinical investigation.

[18]  R. McCarron,et al.  Effects of plasma from hibernating ground squirrels on monocyte-endothelial cell adhesive interactions. , 1997, American journal of physiology. Regulatory, integrative and comparative physiology.

[19]  Trevor Coward,et al.  An In-Vitro Study , 2016 .

[20]  G. R. Ultsch,et al.  Blood viscosity and hematological changes during prolonged submergence in normoxic water of northern and southern musk turtles (Sternotherus odoratus). , 2000, The Journal of experimental zoology.

[21]  R. Henning,et al.  Differential regulation of glomerular and interstitial endothelial nitric oxide synthase expression in the kidney of hibernating ground squirrel. , 2004, Nitric oxide : biology and chemistry.

[22]  David N. Ku,et al.  Determination of Critical Parameters in Platelet Margination , 2012, Annals of Biomedical Engineering.

[23]  D. Reilly,et al.  A Sequence within the Cytoplasmic Tail of GpIIb Independently Activates Platelet Aggregation and Thromboxane Synthesis* , 1998, The Journal of Biological Chemistry.

[24]  M. Hickey,et al.  Evidence of Platelet Activation at Medically Used Hypothermia and Mechanistic Data Indicating ADP as a Key Mediator and Therapeutic Target , 2011, Arteriosclerosis, thrombosis, and vascular biology.

[25]  A. D’Alessandro,et al.  Troubleshooting in platelet storage temperature and new perspectives through proteomics. , 2010, Blood transfusion = Trasfusione del sangue.

[26]  D. Granger,et al.  Platelet–Vessel Wall Interactions in the Microcirculation , 2005, Microcirculation.

[27]  D. Shields,et al.  Monitoring modulators of platelet aggregation in a microtiter plate assay. , 2006, Analytical biochemistry.

[28]  R. Reddick,et al.  Thrombocytopenia of hibernation. Mechanism of induction and recovery. , 1973, Laboratory investigation; a journal of technical methods and pathology.

[29]  M. Hampton,et al.  Circulation and metabolic rates in a natural hibernator: an integrative physiological model. , 2010, American journal of physiology. Regulatory, integrative and comparative physiology.

[30]  A. Toker,et al.  Mechanisms of Cold-induced Platelet Actin Assembly* , 2001, The Journal of Biological Chemistry.

[31]  M. G. Lötter,et al.  Kinetics, Distribution and Sites of Destruction of 111Indium‐labelled Human Platelets , 1980, British journal of haematology.

[32]  J. Fehr,et al.  The hypersplenic spleen. A contractile reservoir of granulocytes and platelets. , 1985, Archives of internal medicine.

[33]  Jian Jin,et al.  AMP Is an Adenosine A1 Receptor Agonist* , 2012, The Journal of Biological Chemistry.

[34]  D. Baruch.  Platelet-Vessel Wall Interactions , 1988, The Bloomsbury Series in Clinical Science.

[35]  S. Morrison,et al.  Central Activation of the A1 Adenosine Receptor (A1AR) Induces a Hypothermic, Torpor-Like State in the Rat , 2013, The Journal of Neuroscience.

[36]  J. F. Storz,et al.  Elucidating Nature's Solutions to Heart, Lung, and Blood Diseases and Sleep Disorders , 2012, Circulation research.

[37]  D. Granger,et al.  Low venular shear rates promote leukocyte-dependent recruitment of adherent platelets. , 2003, American journal of physiology. Gastrointestinal and liver physiology.

[38]  G. Scharbert,et al.  Mild and moderate hypothermia increases platelet aggregation induced by various agonists: a whole blood in vitro study , 2010, Platelets.

[39]  J. Harlan,et al.  Adhesion of Activated Platelets to Endothelial Cells: Evidence for a GPIIbIIIa-dependent Bridging Mechanism and Novel Roles for Endothelial Intercellular Adhesion Molecule 1 (ICAM-1), αvβ3 Integrin, and GPIbα , 1998, The Journal of experimental medicine.

[40]  R. Henning,et al.  The role of endogenous H2S formation in reversible remodeling of lung tissue during hibernation in the Syrian hamster , 2012, Journal of Experimental Biology.

[41]  Sandra L Martin,et al.  Mammalian hibernation: cellular and molecular responses to depressed metabolism and low temperature. , 2003, Physiological reviews.

[42]  D. Devine,et al.  Platelet storage lesion: a new understanding from a proteomic perspective. , 2008, Transfusion medicine reviews.

[43]  F. Geiser,et al.  Metabolic rate and body temperature reduction during hibernation and daily torpor. , 2004, Annual review of physiology.

[44]  H. Morita,et al.  Central A1-receptor activation associated with onset of torpor protects the heart against low temperature in the Syrian hamster. , 2008, American journal of physiology. Regulatory, integrative and comparative physiology.

[45]  K. Drew,et al.  Season Primes the Brain in an Arctic Hibernator to Facilitate Entrance into Torpor Mediated by Adenosine A1 Receptors , 2011, The Journal of Neuroscience.

[46]  J. Hartwig,et al.  Galactosylation does not prevent the rapid clearance of long-term, 4°C-stored platelets , 2008 .

[47]  S. Cash,et al.  The hibernating 13-lined ground squirrel as a model organism for potential cold storage of platelets. , 2012, American journal of physiology. Regulatory, integrative and comparative physiology.

[48]  V. Deutsch,et al.  Megakaryocyte development and platelet production , 2006, British journal of haematology.

[49]  Julio Caballero,et al.  Protective mechanisms of adenosine 5′-monophosphate in platelet activation and thrombus formation , 2013, Thrombosis and Haemostasis.

[50]  J. Vostal,et al.  Liquid cold storage of platelets: a revitalized possible alternative for limiting bacterial contamination of platelet products. , 1997, Transfusion medicine reviews.

[51]  W. B. Sinnamon,et al.  Blood coagulation studies in normothermic, hibernating, and aroused Spermophilus franklini. , 1981, Cryobiology.

[52]  S. Daan,et al.  Normalization of aortic function during arousal episodes in the hibernating ground squirrel. , 2002, Life sciences.

[53]  S. Daan,et al.  Body temperature patterns before, during, and after semi-natural hibernation in the European ground squirrel , 2001, Journal of Comparative Physiology B.

[54]  M. Mangino,et al.  Hibernation confers resistance to intestinal ischemia-reperfusion injury. , 2006, American journal of physiology. Gastrointestinal and liver physiology.

[55]  M. Mangino,et al.  Natural resistance to liver cold ischemia-reperfusion injury associated with the hibernation phenotype. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[56]  R. Henning,et al.  Blood cell dynamics during hibernation in the European Ground Squirrel. , 2010, Veterinary immunology and immunopathology.

[57]  Mehran Ghasemzadeh,et al.  Platelet-leukocyte crosstalk: Linking proinflammatory responses to procoagulant state. , 2013, Thrombosis research.

[58]  Judith N. Mandl,et al.  5′‐AMP impacts lymphocyte recirculation through activation of A2B receptors , 2013, Journal of leukocyte biology.

[59]  S. Lewis,et al.  Spleen function and platelet kinetics. , 1981, Journal of clinical pathology.