Advertisement

Cardiovascular resistance to thrombosis in 13-lined ground squirrels

  • Alison Bonis
  • Leah Anderson
  • Gaëlle Talhouarne
  • Emily Schueller
  • Jenna Unke
  • Catherine Krus
  • Jordan Stokka
  • Anna Koepke
  • Brittany Lehrer
  • Anthony Schuh
  • Jeremiah J. Andersen
  • Scott Cooper
Original Paper
  • 1 Downloads

Abstract

13-lined ground squirrels (Ictidomys tridecemlineatus) enter hibernation as a survival strategy during extreme environmental conditions. Typical ground squirrel hibernation is characterized by prolonged periods of torpor with significantly reduced heart rate, blood pressure, and blood flow, interrupted every few weeks by brief interbout arousals (IBA) during which blood flow fluctuates dramatically. These physiological conditions should increase the risk of stasis-induced blood clots and myocardial ischemia. However, ground squirrels have adapted to survive repeated bouts of torpor and IBA without forming lethal blood clots or sustaining lethal ischemic myocardial damage. The purpose of this study was to determine if ground squirrels are resistant to thrombosis and myocardial ischemia during hibernation. Blood markers of coagulation, fibrinolysis, thrombosis, and ischemia, as well as histological markers of myocardial ischemia were measured throughout the annual hibernation cycle. Hibernating ground squirrels were also treated with isoprenaline to induce myocardial ischemia. Thrombin–antithrombin complex levels were significantly reduced (p < 0.05) during hibernation, while D-dimer level remained unchanged throughout the annual cycle, both consistent with an antithrombotic state. During torpor, the ground squirrels were in a hyperfibrinolytic state with an elevated ratio of tissue plasminogen activator complexed with plasminogen activator inhibitor to total plasminogen activator inhibitor (p < 0.05). Histological markers of myocardial ischemia were reversibly elevated during hibernation with no increase in markers of myocardial cell death in the blood. These data suggest that ground squirrels do not form major blood clots during hibernation through suppression of coagulation and a hyperfibrinolytic state. These animals also demonstrate myocardial resistance to ischemia.

Keywords

Coagulation Fibrinolysis Hemostasis Hibernation Ischemia TAT complex Torpor tPA-PAI1 

Notes

Acknowledgements

We would like to thank Amy Cooper, for her care of the ground squirrels and surgical expertise. This work was supported by grants from the NIH (1R15HL093680) to SC.

Author contributions

SC, AB and LA conceived the ideas and designed the experiments. GT and BL performed the fibrinolysis assays. LA, AB, KC, JU, JJA, and ES performed the myocardial infarction experiments. AB, JS, AS and AK assayed markers of coagulation and fibrinolysis.

Compliance with ethical standards

Conflict of interest

No conflicts of interest, financial or otherwise, are declared by the author(s).

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

References

  1. Albadawi H, Witting AA, Pershad Y, Wallace A, Fleck AR, Hoang P, Khademhosseini A, Oklu R (2017) Animal models of venous thrombosis. Cardiovasc Diagn Therapy 7:S197–S206CrossRefGoogle Scholar
  2. Bertinchant JP, Robert E, Polge A, Marty-Double C, Fabbro-Peray P, Poirey S, Aya G, Juan JM, Ledermann B, de la Coussaye JE, Dauzat M (2000) Comparison of the diagnostic value of cardiac troponin I and T determinations for detecting early myocardial damage and the relationship with histological findings after isoprenaline-induced cardiac injury in rats. Clin Chim Acta 298:13–28CrossRefGoogle Scholar
  3. Bhowmick S, Moore JT, Kirschner DL, Drew KL (2017) Arctic ground squirrel hippocampus tolerates oxygen glucose deprivation independent of hibernation season even when not hibernating and after ATP depletion, acidosis, and glutamate efflux. J Neurochem 142:160–170CrossRefGoogle Scholar
  4. Bogren LK, Olson JM, Carpluk J, Moore JM, Drew KL (2014) Resistance to systemic inflammation and multi organ damage after global ischemia/reperfusion in the arctic ground squirrel. PLoS One 9:e94225CrossRefGoogle Scholar
  5. Buja LM (2005) Myocardial ischemia and reperfusion injury. Cardiovasc Pathol 14:170–175CrossRefGoogle Scholar
  6. Bullard R, Funkhouser G (1962) Estimated regional blood flow by rubidium 86 distribution during arousal from hibernation. Am J Physiol 203:266–270PubMedGoogle Scholar
  7. Byrnes JR, Wolberg AS (2017) New findings on venous thrombogenesis. Hamostaseologie 37:25–35CrossRefGoogle Scholar
  8. Canty JM, Suzuki G (2012) Myocardial perfusion and contraction in acute ischemia and chronic ischemic heart disease. J Mol Cell Cardiol 52:822–831CrossRefGoogle Scholar
  9. Carey HV, Andrews MT, Martin SL (2003) Mammalian hibernation: cellular and molecular responses to depressed metabolism and low temperature. Physiol Rev 83:1153–1181CrossRefGoogle Scholar
  10. Chandra D, Parisini E, Mozaffarian D (2009) Meta-analysis: travel and risk for venous thromboembolism. Ann Intern Med 151:180–190CrossRefGoogle Scholar
  11. Cooper ST, Richters KE, Melin TE, Liu ZJ, Hordyk PJ, Benrud RR, Geiser LR, Cash SE, Simon Shelley C, Howard DR, Ereth MH, Sola-Visner MC (2012) The hibernating 13-lined ground squirrel as a model organism for potential cold storage of platelets. Am J Physiol Regul Integr Comp Physiol 302:R1202–R1208CrossRefGoogle Scholar
  12. Cooper S, Sell S, Nelson L, Hawes J, Benrud JA, Kohlnhofer BM, Burmeister BR, Flood VH (2016a) Von Willebrand factor is reversibly decreased during torpor in 13-lined ground squirrels. J Comp Physiol B 186:131–139CrossRefGoogle Scholar
  13. Cooper ST, Sell SS, Fahrenkrog M, Wilkinson K, Howard DR, Bergen H, Cruz E, Cash SE, Andrews MT, Hampton M (2016b) Effects of hibernation on bone marrow transcriptome in thirteen-lined ground squirrels. Physiol Genomics 120:02015Google Scholar
  14. Cooper S, Lloyd S, Koch A, Lin X, Dobbs K, Theisen T, Zuberbuehler M, Bernhardt K, Gyorfi M, Tenpas T, Hying S, Mortimer S, Lamont C, Lehmann M, Neeves K (2017) Temperature effects on the activity, shape, and storage of platelets from 13-lined ground squirrels. J Comp Physiol B 187:815–825CrossRefGoogle Scholar
  15. de Vrij EL, Vogelaar PC, Goris M, Houwertjes MC, Herwig A, Dugbartey GJ, Boerema AS, Strijkstra AM, Bouma HR, Henning RH (2014) Platelet dynamics during natural and pharmacologically induced torpor and forced hypothermia. PLoS One 9:e93218CrossRefGoogle Scholar
  16. Diaz JA, Obi AT, Myers DD, Wrobleski SK, Henke PK, Mackman N, Wakefield TW (2012) Critical review of mouse models of venous thrombosis. Arterioscler Thromb Vasc Biol 32:556–562CrossRefGoogle Scholar
  17. Douketis J, Tosetto A, Marcucci M, Baglin T, Cushman M, Eichinger S, Palareti G, Poli D, Tait RC, Iorio A (2010) Patient-level meta-analysis: effect of measurement timing, threshold, and patient age on ability of D-dimer testing to assess recurrence risk after unprovoked venous thromboembolism. Ann Intern Med 153:523–531CrossRefGoogle Scholar
  18. Emerson TE Jr, Fournel MA, Redens TB, Taylor FB Jr (1989) Efficacy of antithrombin III supplementation in animal models of fulminant Escherichia coli endotoxemia or bacteremia. Am J Med 87:27S–33SCrossRefGoogle Scholar
  19. Esmon CT (2009) Basic mechanisms and pathogenesis of venous thrombosis. Blood Rev 23:225–229CrossRefGoogle Scholar
  20. Frerichs K, Dienel G, Cruz N, Sokoloff L, Hallenbeck J (1995) Rates of glucose utilization in brain of active and hibernating ground squirrels. Am J Physiol 268:R445–R453PubMedGoogle Scholar
  21. Friedrich AU, Kakuturu J, Schnorr PJ, Beyer DE Jr, Palesty JA, Dickson EW, Basadonna G, Cahan MA (2017) Comparative coagulation studies in hibernating and summer-active black bears (Ursus americanus). Thromb Res 158:16–18CrossRefGoogle Scholar
  22. Ganote CE (1983) Contraction band necrosis and irreversible myocardial injury. J Mol Cell Cardiol 15:67–73CrossRefGoogle Scholar
  23. Hampton M, Nelson BT, Andrews MT (2010) Circulation and metabolic rates in a natural hibernator: an integrative physiological model. Am J Physiol Regul Integr Comp Physiol 299:R1478–R1488CrossRefGoogle Scholar
  24. Hampton M, Melvin RG, Andrews MT (2013) Transcriptomic analysis of brown adipose tissue across the physiological extremes of natural hibernation. PLoS One 8:e85157CrossRefGoogle Scholar
  25. Harada N, Okajima K, Kushimoto S, Isobe H, Tanaka K (1999) Antithrombin reduces ischemia/reperfusion injury of rat liver by increasing the hepatic level of prostacyclin. Blood 93:157–164PubMedGoogle Scholar
  26. Jani A, Orlicky DJ, Karimpour-Fard A, Epperson LE, Russell RL, Hunter LE, Martin SL (2012) Kidney proteome changes provide evidence for a dynamic metabolism and regional redistribution of plasma proteins during torpor-arousal cycles of hibernation. Physiol Genom 44:717–727CrossRefGoogle Scholar
  27. Jochum M (1995) Influence of high-dose antithrombin concentrate therapy on the release of cellular proteinases, cytokines, and soluble adhesion molecules in acute inflammation. Semin Hematol 32:19–32PubMedGoogle Scholar
  28. Kurtz CC, Lindell SL, Mangino MJ, Carey HV (2006) Hibernation confers resistance to intestinal ischemia-reperfusion injury. Am J Physiol Gastrointest Liver Physiol 291:G895–G901CrossRefGoogle Scholar
  29. Lau CL, Zhao Y, Kim J, Kron IL, Sharma A, Yang Z, Laubach VE, Linden J, Ailawadi G, Pinsky DJ (2009) Enhanced fibrinolysis protects against lung ischemia-reperfusion injury. J Thorac Cardiovasc Surg 137:1241–1248CrossRefGoogle Scholar
  30. Lechler E, Penick GD (1963) Blood clotting defect in hibernating ground squirrels (Citellus tridecemlineatus). Am J Physiol 205:985–988PubMedGoogle Scholar
  31. Lee SY, Niikura T, Iwakura T, Sakai Y, Kuroda R, Kurosaka M (2017) Thrombin-antithrombin III complex tests. J Orthop Surg (Hong Kong) 25:170840616684501Google Scholar
  32. Lindenblatt N, Menger MD, Klar E, Vollmar B (2005) Sustained hypothermia accelerates microvascular thrombus formation in mice. Am J Physiol Heart Circ Physiol 289:H2680–H2687CrossRefGoogle Scholar
  33. Lyman CP, O’brien RC (1960) Circulatory changes in the thirteen-lined ground squirrel during the hibernation cycle. Bull Mus Comp Zool 124:353–372Google Scholar
  34. Ma YL, Zhu X, Rivera PM, Toien O, Barnes BM, LaManna JC, Smith MA, Drew KL (2005) Absence of cellular stress in brain after hypoxia induced by arousal from hibernation in Arctic ground squirrels. Am J Physiol Regul Integr Comp Physiol 289:R1297–R1306CrossRefGoogle Scholar
  35. McArthur MD, Milsom WK (1991) Changes in ventilation and respiratory sensitivity associated with hibernation in Columbian (Spermophilus columbianus) and golden-mantled (Spermophilis lateralis) ground squirrels. Physiol Zool 64:940–959CrossRefGoogle Scholar
  36. Mythili S, Malathi N (2015) Diagnostic markers of acute myocardial infarction. Biomed Rep 3:743–748CrossRefGoogle Scholar
  37. Nelson CJ, Otis JP, Carey HV (2009) A role for nuclear receptors in mammalian hibernation. J Physiol 587:1863–1870CrossRefGoogle Scholar
  38. Ohkura N, Shirakura M, Nakatani E, Oishi K, Atsumi G-i (2012) Associations between plasma PAI-1 concentrations and its expressions in various organs in obese model mice. Thromb Res 130:e301–e304CrossRefGoogle Scholar
  39. Otis JP, Ackermann LW, Denning GM, Carey HV (2010) Identification of qRT-PCR reference genes for analysis of opioid gene expression in a hibernator. J Comp Physiol B 180:619–629CrossRefGoogle Scholar
  40. Otis JP, Pike AC, Torrealba JR, Carey HV (2017) Hibernation reduces cellular damage caused by warm hepatic ischemia-reperfusion in ground squirrels. J Comp Physiol B 187:639–648CrossRefGoogle Scholar
  41. Pinsky DJ, Liao H, Lawson CA, Yan SF, Chen J, Carmeliet P, Loskutoff DJ, Stern DM (1998) Coordinated induction of plasminogen activator inhibitor-1 (PAI-1) and inhibition of plasminogen activator gene expression by hypoxia promotes pulmonary vascular fibrin deposition. J Clin Invest 102:919–928CrossRefGoogle Scholar
  42. Pivorun EB, Sinnamon WB (1981) Blood coagulation studies in normothermic, hibernating, and aroused Spermophilus franklini. Cryobiology 18:515–520CrossRefGoogle Scholar
  43. Prendergast BJ, Freeman DA, Zucker I, Nelson RJ (2002) Periodic arousal from hibernation is necessary for initiation of immune responses in ground squirrels. Am J Physiol Regul Integr Comp Physiol 282:R1054–R1062CrossRefGoogle Scholar
  44. Pulivarthi S, Gurram MK (2014) Effectiveness of D-dimer as a screening test for venous thromboembolism: an update. N Am J Med Sci 6:491–499CrossRefGoogle Scholar
  45. Quinones QJ, Zhang Z, Ma Q, Smith MP, Soderblom E, Moseley MA, Bain J, Newgard CB, Muehlbauer MJ, Hirschey M, Drew KL, Barnes BM, Podgoreanu MV (2016) Proteomic profiling reveals adaptive responses to surgical myocardial ischemia-reperfusion in hibernating arctic ground squirrels compared to rats. Anesthesiology 124:1296–1310CrossRefGoogle Scholar
  46. Reddick RL, Poole BL, Penick GD (1973) Thrombocytopenia of hibernation. Mechanism of induction and recovery. Lab Invest 28:270–278PubMedGoogle Scholar
  47. Saleem TS, Lokanath N, Prasanthi A, Madhavi M, Mallika G, Vishnu MN (2013) Aqueous extract of Saussurea lappa root ameliorate oxidative myocardial injury induced by isoproterenol in rats. J Adv Pharm Technol Res 4:94–100CrossRefGoogle Scholar
  48. Salzman MM, Cheng Q, Deklotz RJ, Dulai GK, Douglas HF, Dikalova AE, Weihrauch D, Barnes BM, Riess ML (2017) Lipid emulsion enhances cardiac performance after ischemia-reperfusion in isolated hearts from summer-active arctic ground squirrels. J Comp Physiol B 187:715–724CrossRefGoogle Scholar
  49. Sarkar K, Cai Z, Gupta R, Parajuli N, Fox-Talbot K, Darshan MS, Gonzalez FJ, Semenza GL (2012) Hypoxia-inducible factor 1 transcriptional activity in endothelial cells is required for acute phase cardioprotection induced by ischemic preconditioning. Proc Natl Acad Sci USA 109:10504–10509CrossRefGoogle Scholar
  50. Scheer FA, Shea SA (2014) Human circadian system causes a morning peak in prothrombotic plasminogen activator inhibitor-1 (PAI-1) independent of the sleep/wake cycle. Blood 123:590–593CrossRefGoogle Scholar
  51. Scheer FA, Michelson AD, Frelinger AL III, Evoniuk H, Kelly EE, McCarthy M, Doamekpor LA, Barnard MR, Shea SA (2011) The human endogenous circadian system causes greatest platelet activation during the biological morning independent of behaviors. PLoS One 6:e24549CrossRefGoogle Scholar
  52. Siddiqui M, Ahmad U, Khan A, Ahmad M, Badruddeen Khalid M, Akhtar J (2016) Isoprenaline: a tool for inducing myocardial infarction in experimental animals. Int J Res Rev Pharm Appl Sci 6:1318–1326Google Scholar
  53. Smith DE, Lewis YS, Svihla G (1954) Prolongation of clotting time in the dormant bat (Myotis lucifugus). Experientia 10:218CrossRefGoogle Scholar
  54. Stampfli SF, Camici GG, Keller S, Rozenberg I, Arras M, Schuler B, Gassmann M, Garcia I, Luscher TF, Tanner FC (2014) Restraint stress enhances arterial thrombosis in vivo—role of the sympathetic nervous system. Stress 17:126–132CrossRefGoogle Scholar
  55. Suomalainen P, Lehto E (1952) Prolongation of clotting time in hibernation. Experientia 8:65CrossRefGoogle Scholar
  56. Svihla A, Bowman H, Pearson R (1952a) Prolongation of blood clotting time in the dormant hamster. Science 115:272CrossRefGoogle Scholar
  57. Svihla A, Bowman H, Ritenour R (1952b) Relation of prothrombin to the prolongation of clotting time in aestivating ground squirrels. Science 115:306–307CrossRefGoogle Scholar
  58. Svihla A, Bowman H, Ritenour R (1953) Stimuli and their effects on awakening of dormant ground squirrels. Am J Physiol 172:681–683PubMedGoogle Scholar
  59. van Adrichem RA, Debeij J, Nelissen RG, Schipper IB, Rosendaal FR, Cannegieter SC (2014) Below-knee cast immobilization and the risk of venous thrombosis: results from a large population-based case-control study. J Thromb Haemost 12:1461–1469CrossRefGoogle Scholar
  60. von Brühl L, Stark K, Steinhart A, Chandraratne S, Konrad I, Lorenz M, Khandoga A, Tirniceriu A, Coletti R, Köllnberger M, Byrne R, Laitinen I, Walch A, Brill A, Pfeiler S, Manukyan D, Braun S, Lange P, Riegger J, Ware J, Eckart A, Haidari S, Rudelius M, Schulz C, Echtler K, Brinkmann V, Schwaiger M, Preissner K, Wagner D, Mackman N, Engelmann B, Massberg S (2012) Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo. J Exp Med 209:819–835CrossRefGoogle Scholar
  61. Wang J, Wang Y, Wang J, Gao J, Tong C, Manithody C, Li J, Rezaie AR (2013) Antithrombin is protective against myocardial ischemia and reperfusion injury. J Thromb Haemost 11:1020–1028CrossRefGoogle Scholar
  62. Watanabe R, Wada H, Miura Y, Murata Y, Watanabe Y, Sakakura M, Okugawa Y, Nakasaki T, Mori Y, Nishikawa M, Gabazza EC, Shiku H, Nobori T (2001) Plasma levels of total plasminogen activator inhibitor-I (PAI-I) and tPA/PAI-1 complex in patients with disseminated intravascular coagulation and thrombotic thrombocytopenic purpura. Clin Appl Thromb Hemost 7:229–233CrossRefGoogle Scholar
  63. Wei B, You MG, Ling JJ, Wei LL, Wang K, Li WW, Chen T, Du QM, Ji H (2013) Regulation of antioxidant system, lipids and fatty acid beta-oxidation contributes to the cardioprotective effect of sodium tanshinone IIA sulphonate in isoproterenol-induced myocardial infarction in rats. Atherosclerosis 230:148–156CrossRefGoogle Scholar
  64. Weymann A, Sabashnikov A, Ali-Hasan-Al-Saegh S, Popov AF, Jalil Mirhosseini S, Baker WL, Lotfaliani M, Liu T, Dehghan H, Yavuz S, de Oliveira Sa MP, Jang JS, Zeriouh M, Meng L, D’Ascenzo F, Deshmukh AJ, Biondi-Zoccai G, Dohmen PM, Calkins H, Cardiac S, Cardiology-Group Imcsc-Group, IM (2017) Predictive role of coagulation, fibrinolytic, and endothelial markers in patients with atrial fibrillation, stroke, and thromboembolism: a meta-analysis, meta-regression, and systematic review. Med Sci Monit Basic Res 23: 97–140PubMedPubMedCentralGoogle Scholar
  65. Zatzman ML (1984) Renal and cardiovascular effects of hibernation and hypothermia. Cryobiology 21:593–614CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Alison Bonis
    • 1
  • Leah Anderson
    • 1
  • Gaëlle Talhouarne
    • 1
  • Emily Schueller
    • 1
  • Jenna Unke
    • 1
  • Catherine Krus
    • 1
  • Jordan Stokka
    • 1
  • Anna Koepke
    • 1
  • Brittany Lehrer
    • 1
  • Anthony Schuh
    • 1
  • Jeremiah J. Andersen
    • 2
  • Scott Cooper
    • 1
  1. 1.Biology DepartmentUniversity of Wisconsin-La CrosseLa CrosseUSA
  2. 2.Gundersen Health SystemLa CrosseUSA

Personalised recommendations