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Temperature effects on the activity, shape, and storage of platelets from 13-lined ground squirrels

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Abstract

The objective of this study is to determine how a hibernating mammal avoids the formation of blood clots under periods of low blood flow. A microfluidic vascular injury model was performed to differentiate the effects of temperature and shear rate on platelet adhesion to collagen. Human and ground squirrel whole blood was incubated at 15 or 37 °C and then passed through a microfluidic chamber over a 250-µm strip of type I fibrillar collagen at that temperature and the shear rates of 50 or 300 s−1 to simulate torpid and aroused conditions, respectively. At 15 °C, both human and ground squirrel platelets showed a 90–95% decrease in accumulation on collagen independent of shear rate. At 37 °C, human platelet accumulation reduced by 50% at 50 s−1 compared to 300 s−1, while ground squirrel platelet accumulation dropped by 80%. When compared to platelets from non-hibernating animals, platelets from animals collected after arousal from torpor showed a 60% decrease in binding at 37 °C and 300 s−1, but a 2.5-fold increase in binding at 15 °C and 50 s−1. vWF binding in platelets from hibernating ground squirrels was decreased by 50% relative to non-hibernating platelets. The source of the plasma that platelets were stored in did not affect the results indicating that the decreased vWF binding was a property of the platelets. Upon chilling, ground squirrel platelets increase microtubule assembly leading to the formation of long rods. This shape change is concurrent with sequestration of platelets in the liver and not the spleen. In conclusion, it appears that ground squirrel platelets are sequestered in the liver during torpor and have reduced binding capacity for plasma vWF and lower accumulation on collagen at low shear rates and after storage at cold temperatures, while still being activated by external agonists. These adaptations would protect the animals from spontaneous thrombus formation during torpor but allow them to restore normal platelet function upon arousal.

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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 S.C. and NSF CAREER (CBET-1351672), American Heart Association (14GRNT20410094), and the National Institutes of Health (R01HL120728, R21NS082933) to K.N. S.L. and C.L. received a UW-La Crosse Dean’s Distinguished Undergraduate Summer fellowship.

Author contributions

S.C. and K.N. conceived the ideas and designed the experiments. A.K. and M.Z. performed the platelet vWF and fibrinogen binding assays. S.L., K.D., T. Theisen, and M.L. performed microfluidics. M.G. and T. Tenpas performed immunohistochemistry. X.L., K.B., and S.H. measured microtubule kinetics. S.M. and C.L. performed splenectomy experiments.

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Authors and Affiliations

  1. Biology Department, University of Wisconsin-La Crosse, 1725 State St., La Crosse, WI, 54601, USA

    Scott Cooper, Sarah Lloyd, Anthony Koch, Xingxing Lin, Katie Dobbs, Thomas Theisen, Matt Zuberbuehler, Kaley Bernhardt, Michael Gyorfi, Tanner Tenpas, Skyler Hying, Sarah Mortimer & Christine Lamont

  2. Chemical and Biological Engineering Department, Colorado School of Mines, Golden, CO, 80401, USA

    Marcus Lehmann & Keith Neeves

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  1. Scott Cooper
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Correspondence to Scott Cooper.

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

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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.

Additional information

Communicated by F. Breukelen.

This manuscript is part of the special issue Hibernation—Guest Editors: Frank van Breukelen and Jenifer C. Utz.

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Cooper, S., Lloyd, S., Koch, A. et al. Temperature effects on the activity, shape, and storage of platelets from 13-lined ground squirrels. J Comp Physiol B 187, 815–825 (2017). https://doi.org/10.1007/s00360-017-1081-x

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