Anoxia-tolerant hepatocytes: Model system for study of reversible metabolic suppression

L. T. Buck, S. C. Land, P. W. Hochachka

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Chrysemis picta bellii is well known for its ability to survive extended anoxic periods and has been widely used as a model system to study anoxic metabolism. Described here is a method for the isolation of anoxia-tolerant hepatocytes from this species. Freshly isolated hepatocytes were determined to be viable based on trypan blue exclusion, gluconeogenic capacity from [14C]lactate, responsiveness to epinephrine and glucagon, and maintenance of cellular adenylate concentrations. Under anoxic conditions for 10 h there was no significant increase in cell staining and no decrease in cellular ATP concentration. Furthermore, the addition of cyanide at the 5-h mark did not result in any significant differences in these parameters; however, iodoacetate added at this time caused trypan blue staining to increase and ATP concentrations to fall. The rate of glucose production from the cells was threefold greater under anoxic than normoxic conditions, underscoring the important role of the liver in supplying substrate during anoxia. From the rate of O2 consumption and rate of lactate production under anaerobic conditions, ATP turnover rates were calculated to be 68.4 ± 7.2 and 6.5 ± 0.43 μmol ATP · g-1 · h-1, respectively; this corresponds to a 90% decrease in metabolic rate during anoxia. Within a cellular system such as this the more complex regulatory mechanisms involved in a large coordinated reduction in metabolism can be probed.

Original languageEnglish
Pages (from-to)R49-R56
Number of pages8
JournalAmerican Journal of Physiology: Regulatory Integrative and Comparative Physiology
Volume265
Issue number1
DOIs
Publication statusPublished - 1 Jul 1993

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Hepatocytes
Adenosine Triphosphate
Trypan Blue
Lactic Acid
Staining and Labeling
Iodoacetates
Cyanides
Glucagon
Epinephrine
Maintenance
Glucose
Hypoxia
Liver

Keywords

  • adenosine 5'-triphosphate turnover
  • Chrysemys picta bellii
  • coordinated metabolic depression
  • turtle

Cite this

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abstract = "Chrysemis picta bellii is well known for its ability to survive extended anoxic periods and has been widely used as a model system to study anoxic metabolism. Described here is a method for the isolation of anoxia-tolerant hepatocytes from this species. Freshly isolated hepatocytes were determined to be viable based on trypan blue exclusion, gluconeogenic capacity from [14C]lactate, responsiveness to epinephrine and glucagon, and maintenance of cellular adenylate concentrations. Under anoxic conditions for 10 h there was no significant increase in cell staining and no decrease in cellular ATP concentration. Furthermore, the addition of cyanide at the 5-h mark did not result in any significant differences in these parameters; however, iodoacetate added at this time caused trypan blue staining to increase and ATP concentrations to fall. The rate of glucose production from the cells was threefold greater under anoxic than normoxic conditions, underscoring the important role of the liver in supplying substrate during anoxia. From the rate of O2 consumption and rate of lactate production under anaerobic conditions, ATP turnover rates were calculated to be 68.4 ± 7.2 and 6.5 ± 0.43 μmol ATP · g-1 · h-1, respectively; this corresponds to a 90{\%} decrease in metabolic rate during anoxia. Within a cellular system such as this the more complex regulatory mechanisms involved in a large coordinated reduction in metabolism can be probed.",
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Anoxia-tolerant hepatocytes : Model system for study of reversible metabolic suppression. / Buck, L. T.; Land, S. C.; Hochachka, P. W.

In: American Journal of Physiology: Regulatory Integrative and Comparative Physiology, Vol. 265, No. 1 , 01.07.1993, p. R49-R56.

Research output: Contribution to journalArticle

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