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Von Willebrand factor is reversibly decreased during torpor in 13-lined ground squirrels

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Abstract

During torpor in a hibernating mammal, decreased blood flow increases the risk of blood clots such as deep vein thrombi (DVT). In other animal models platelets, neutrophils, monocytes and von Willebrand factor (VWF) have been found in DVT. Previous research has shown that hibernating mammals decrease their levels of platelets and clotting factors VIII (FVIII) and IX (FIX), increasing both bleeding time and activated partial thromboplastin time. In this study, FVIII, FIX and VWF activities and mRNA levels were measured in torpid and non-hibernating ground squirrels (Ictidomys tridecemlineatus). Here, we show that VWF high molecular weight multimers, collagen-binding activity, lung mRNA and promoter activity decrease during torpor. The VWF multimers reappear in plasma within 2 h of arousal in the spring. Similarly, FIX activity and liver mRNA both dropped threefold during torpor. In contrast, FVIII liver mRNA levels increased twofold while its activity dropped threefold, consistent with a post-transcriptional decrease in FVIII stability in the plasma due to decreased VWF levels. Finally, both neutrophils and monocytes are decreased eightfold during torpor which could slow the formation of DVT. In addition to providing insight in how blood clotting can be regulated to allow mammals to survive in extreme environments, hibernating ground squirrels provide an interesting model for studying.

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Acknowledgments

We would like to thank Amy Cooper, for her care of the ground squirrels and surgical expertise. We thank Dana Vaughan for helping us establish our breeding colony of ground squirrels. J. Evan Sadler and Alisa Wolberg provided critical review of the paper. Jeffrey Wren, Brian Kleinschmidt, and Elizabeth Klatt assisted with the VWF immunoblots. This work was supported by grants from the NIH (1R15HL093680, to S.T.C. and 1K08HL102260 to V.H.F), UW-System WiTAG (to J.A.B.), and an UW-La Crosse faculty development grant to S.T.C. B.M.K. and S.S. received a UW-La Crosse Dean’s Distinguished Undergraduate Summer fellowship. J.A.B. received an NSF-REU summer fellowship.

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Authors and Affiliations

  1. University of Wisconsin-La Crosse, La Crosse, WI, USA

    Scott Cooper, Shawn Sell, Luke Nelson, Jennifer Hawes, Jacob A. Benrud, Bridget M. Kohlnhofer & Bradley R. Burmeister

  2. Medical College of Wisconsin, Milwaukee, WI, 53226, USA

    Veronica H. Flood

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  1. Scott Cooper
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Correspondence to Scott Cooper.

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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.

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Communicated by H.V. Carey.

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360_2015_941_MOESM1_ESM.tif

Supplemental Fig. 1. Sections of brain from ground squirrels. Sections were fixed, paraffin embedded, and 4 µm sections made using a microtome. Panels A and D show creases that are artifacts of sectioning. Slides were stained with hematoxylin and eosin, mounted and images taken on a Nikon Eclipse 80i microscope and DS-Ri1 camera using 10X and 40X objectives. A and E are from a July non-hibernating squirrel, B and F are from a January torpid animal, C and G are from a January IBA animal, D and H are from a spring animal 2 h post-arousal. The blood that is present in the vessels is a feature of port-mortem coagulation, rather than thrombosis/embolism since no organized fibrin networks can be found (TIFF 2836 kb)

360_2015_941_MOESM2_ESM.tif

Supplemental Fig. 2. Sections of lung from ground squirrels. Sections were fixed, paraffin embedded, and 4 µm sections made using a microtome. Slides were stained with hematoxylin and eosin, mounted and images taken on a Nikon Eclipse 80i microscope and DS-Ri1 camera using 10X and 40X objectives. A and E are from a July non-hibernating squirrel, B and F are from a January torpid animal, C and G are from a January IBA animal, D and H are from a spring animal 2 h post-arousal. The blood that is present in the vessels is a feature of port-mortem coagulation, rather than thrombosis/embolism since no organized fibrin networks can be found (TIFF 3139 kb)

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Cooper, S., Sell, S., Nelson, L. et al. Von Willebrand factor is reversibly decreased during torpor in 13-lined ground squirrels. J Comp Physiol B 186, 131–139 (2016). https://doi.org/10.1007/s00360-015-0941-5

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