Journal of Comparative Physiology B

, Volume 189, Issue 2, pp 199–211 | Cite as

Contractile function of the excised hagfish heart during anoxia exposure

  • L. A. Gatrell
  • E. Farhat
  • W. G. Pyle
  • Todd E. GillisEmail author
Original Paper


Pacific hagfish, Eptatretus stoutii, can recover from 36 h of anoxia and their systemic hearts continue to work throughout the exposure. Recent work demonstrates that glycogen stores are utilized in the E. stoutii heart during anoxia but that these are not sufficient to support the measured rate of ATP production. One metabolic fuel that could supplement glycogen during anoxia is glycerol. This substrate can be derived from lipid stores, stored in the heart, or delivered via the blood. The purpose of this study was to determine the effect of glycerol on the contractile function of the excised E. stoutii heart during anoxia exposure. When excised hearts, perfused with metabolite free saline (mf-saline), were exposed to anoxia for 12 h, there was no difference in heart rate, pressure generation (max-dP), rate of contraction (max-dP/dtsys), or rate of relaxation (max-dP/dtdia) compared to hearts perfused with mf-saline in normoxia. However, hearts perfused with saline containing glycerol (gly-saline) in anoxia had higher max-dP, max-dP/dtsys, and max-dP/dtdia than hearts perfused with mf-saline in anoxia. Tissue levels of glycerol increased when hearts were perfused with gly-saline in normoxia, but not when perfused with gly-saline in anoxia. Anoxia exposure did not affect the activities of triglyceride lipase, glycerol kinase, or glycerol-3-phosphate dehydrogenase. This study suggests that glycerol stimulates cardiac function in the hagfish but that it is not derived from stored lipids. How glycerol may stimulate contraction is not known. This could be as an energy substrate, as an allosteric factor, or a combination of the two.


Contractile function Anaerobic metabolism Pressure generation Working heart preparation Glycerol 



The Authors would like to thank C.R. Freedman and Dr. D.S. Fudge for comments on an earlier draft, as well as Dr. I. Lorenzen-Schmidt and A. Pierce for technical assistance.


This work was supported by an operating grant from the Canadian Institutes of Health Research to W.G.P, and a Discovery Grant, and Discovery Accelerator Supplement, from the National Sciences and Engineering Research Council of Canada to T.E.G.


  1. Areta JL, Hopkins WG (2018) Skeletal muscle glycogen content at rest and during endurance exercise in humans: a meta-analysis. Sports Med. Google Scholar
  2. Bergmeyer HU, Bergmeyer Jr, Grassl M (1983) Methods of enzymatic analysis, 3rd edn. Verlag Chemie, WeinheimGoogle Scholar
  3. Bucking C, Glover CN, Wood CM (2011) Digestion under duress: nutrient acquisition and metabolism during hypoxia in the Pacific hagfish. Physiol Biochem Zool 84(6):607–617. CrossRefGoogle Scholar
  4. Cox GK, Sandblom E, Farrell AP (2010) Cardiac responses to anoxia in the Pacific hagfish, Eptatretus stoutii. J Exp Biol 213(21):3692–3698. doi: CrossRefGoogle Scholar
  5. Cox GK, Sandblom E, Richards JG, Farrell AP (2011) Anoxic survival of the Pacific hagfish (Eptatretus stoutii). J Comp Physiol B Biochem Syst Environ Physiol 181(3):361–371. CrossRefGoogle Scholar
  6. Crabtree B, Newsholme EA (1972) The activities of phosphorylase, hexokinase, phosphofructokinase, lactate dehydrogenase and the glycerol 3-phosphate dehydrogenases in muscles from vertebrates and invertebrates. Biochem J 126(1):49–58CrossRefGoogle Scholar
  7. Cutler CP (2006) Cloning of an aquaporin 9 gene orthologue from the hagfish (Myxine glutinosa). Bull Mt Desert Isl Biol Lab 45:42–43Google Scholar
  8. Dhar-Chowdhury P, Harrell MD, Han SY, Jankowska D, Parachuru L, Morrissey A, Srivastava S, Liu W, Malester B, Yoshida H, Coetzee WA (2005) The glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase, triose-phosphate isomerase, and pyruvate kinase are components of the K(ATP) channel macromolecular complex and regulate its function. J Biol Chem 280(46):38464–38470. CrossRefGoogle Scholar
  9. Driedzic WR, Ewart KV (2004) Control of glycerol production by rainbow smelt (Osmerus mordax) to provide freeze resistance and allow foraging at low winter temperatures. Comp Biochem Physiol B Biochem Mol Biol 139(3):347–357. CrossRefGoogle Scholar
  10. Driedzic WR, West JL, Sephton DH, Raymond JA (1998) Enzyme activity levels associated with the production of glycerol as an antifreeze in liver of rainbow smelt (Osmerus mordax). Fish Physiol Biochem 18(2):125–134. CrossRefGoogle Scholar
  11. Farrell AP, Smith F (2017) Cardiac form, function and physiology. In: Gamperl A, Gillis TE, Farrell AP, Brauner CJ (eds) The cardiovascular syste, form function and control, vol 36A. Elsivier: New York pp 155–264CrossRefGoogle Scholar
  12. Farrell A, Gamperl A, Hicks J, Shiels H, Jain K (1996) Maximum cardiac performance of rainbow trout (Oncorhynchus mykiss) at temperatures approaching their upper lethal limit. J Exp Biol 199(Pt 3):663–672Google Scholar
  13. Forster ME, Axelsson M, Farrell AP, Nilsson S (1991) Cardiac-function and circulation in hagfishes. Can J Zool 69(7):1985–1992. doi: CrossRefGoogle Scholar
  14. Gambert S, Helies-Toussaint C, Grynberg A (2007) Extracellular glycerol regulates the cardiac energy balance in a working rat heart model. Am J Physiol Heart Circ Physiol 292(3):H1600–H1606. CrossRefGoogle Scholar
  15. Gillis TE, Regan MD, Cox GK, Harter TS, Brauner CJ, Richards JG, Farrell AP (2015) Characterizing the metabolic capacity of the anoxic hagfish heart. J Exp Biol 218:3754–3761. CrossRefGoogle Scholar
  16. Graham MS, Farrell AP (1989) The effect of temperature acclimation and adrenaline on the performance of a perfused trout heart. Physiol Zool 62:38–61CrossRefGoogle Scholar
  17. Hansen CA, Sidell BD (1983) Atlantic hagfish cardiac muscle: metabolic basis of tolerance to anoxia. Am J Physiol 244:R356–R362CrossRefGoogle Scholar
  18. Herr JE, Clifford AM, Goss GG, Fudge DS (2014) Defensive slime formation in Pacific hagfish requires Ca2+- and aquaporin-mediated swelling of released mucin vesicles. J Exp Biol 217(Pt 13):2288–2296. CrossRefGoogle Scholar
  19. Hibuse T, Maeda N, Nakatsuji H, Tochino Y, Fujita K, Kihara S, Funahashi T, Shimomura I (2009) The heart requires glycerol as an energy substrate through aquaporin 7, a glycerol facilitator. Cardiovasc Res 83(1):34–41. CrossRefGoogle Scholar
  20. Icardo JM, Colvee E, Schorno S, Lauriano ER, Fudge DS, Glover CN, Zaccone G (2016) Morphological analysis of the hagfish heart. I. The ventricle, the arterial connection and the ventral aorta. J Morphol 277(3):326–340. CrossRefGoogle Scholar
  21. Inui Y, Yu JY, Gorbman A (1978) Effect of bovine insulin on the incorporation of [14C]glycine into protein and carbohydrate in liver and muscle of hagfish, Eptatretus stouti. General Comp Endocrinol 36(1):133–141CrossRefGoogle Scholar
  22. Johansen KL (1960) Circulation in the hagfish, Myxine glutinosa. Biol Bull, 118:298CrossRefGoogle Scholar
  23. Klaiman JM, Pyle WG, Gillis TE (2014) Cold acclimation increases cardiac myofilament function and ventricular pressure generation in trout. J Exp Biol 217(Pt 23):4132–4140. CrossRefGoogle Scholar
  24. Martini FH (1998) The ecology of hagfishes. In: Jørgensen JM, Lomholt JP, Weber RE, Malte H (eds) Biology of hagfishes. Springer Nature, Switzerland, pp 57–77CrossRefGoogle Scholar
  25. Mendonca PC, Genge AG, Deitch EJ, Gamperl AK (2007) Mechanisms responsible for the enhanced pumping capacity of the in situ winter flounder heart (Pseudopleuronectes americanus). Am J Physiol Regul Integr Comp Physiol 293(5):R2112–R2119. CrossRefGoogle Scholar
  26. Perez-Jimenez A, Hidalgo MC, Morales AE, Arizcun M, Abellan E, Cardenete G (2009) Use of different combinations of macronutrients in diets for dentex (Dentex dentex): effects on intermediary metabolism. Comp Biochem Physiol A Mol Integr Physiol 152(3):314–321. CrossRefGoogle Scholar
  27. Robertson JD (1976) Chemical composition of body-fluids and muscle of hagfish Myxine-glutinosa and rabbit-fish Chimaera-monstrosa. J Zool 178(Feb):261–277Google Scholar
  28. Ryan NM (1996) Subcellular fractionation of animal tissues. Methods Mol Biol 59:49–56. Google Scholar
  29. Satchell GH (1991) Physiology and form of fish circulation. Cambridge University Press, CambridgeCrossRefGoogle Scholar
  30. Sidell BD, Beland KF (1980) Lactate dehydrogenases of Atlantic hagfish: physiological and evolutionary implications of a primitive heart isozyme. Science 207(4432):769–770CrossRefGoogle Scholar
  31. Treberg JR, Wilson CE, Richards RC, Ewart KV, Driedzic WR (2002) The freeze-avoidance response of smelt Osmerus mordax: initiation and subsequent suppression of glycerol, trimethylamine oxide and urea accumulation. J Exp Biol 205(10):1419–1427Google Scholar
  32. van der Vusse GJ, Glatz JF, Stam HC, Reneman RS (1992) Fatty acid homeostasis in the normoxic and ischemic heart. Physiol Rev 72(4):881–940CrossRefGoogle Scholar
  33. Wakayama JE, Swanson JR (1977) Ultraviolet spectrometry of serum triglycerides by a totally enzymic method adapted to a centrifugal analyzer. Clin Chem 23(2 PT. 1):223–228Google Scholar
  34. Walsh PJ, Foster GD, Moon TW (1983) The effects of temperature on metabolism of the American eel Anguilla-rostrata (Lesueur)—compensation in the summer and torpor in the winter. Physiol Zool 56(4):532–540. doi: CrossRefGoogle Scholar
  35. Wilson CM, Roa JN, Cox GK, Tresguerres M, Farrell AP (2016) Introducing a novel mechanism to control heart rate in the ancestral Pacific hagfish. J Exp Biol 219(Pt 20):3227–3236. CrossRefGoogle Scholar
  36. Wittels B, Spann JF Jr (1968) Defective lipid metabolism in the failing heart. J Clin Investig 47(8):1787–1794. CrossRefGoogle Scholar
  37. Young DA, King DS, Chen M, Norris B, Nemeth PM (1988) A novel method for measurement of triglyceride lipase activity: suitable for microgram and nanogram quantities of tissue. J Lipid Res 29:527–532Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Integrative BiologyUniversity of GuelphGuelphCanada
  2. 2.Department of BiologyUniversity of OttawaOttawaCanada
  3. 3.Department of Biomedical SciencesUniversity of GuelphGuelphCanada

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