Perifused precision-cut liver slice system for the study of hormone-regulated hepatic glucose metabolism

Roberta L. Mckee, Victor J Hruby, Devendra B. Trivedi, David G Johnson, A Jay Gandolfi, Carlos L. Krumdieck, Klaus Brendel

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A nonrecirculatory perifusion system for precision-cut rat liver slices has been developed and utilized for investigating hormone-regulated hepatic glucose metabolism. In this system, slices are cultured in a highly controlled environment and exhibit excellent retention of viability as judged by their maintenance of intracellular potassium and glycogen contents. Using this system, the complex physiological phenomenon of hormone-regulated glycogenolysis was investigated at both extra- and intracellular sites. Specifically, the sensitive responses of intracellular cyclic AMP (cAMP) production, activation of cyclic AMP-dependent protein kinase, and production of glucose upon glucagon stimulation have been measured. The maximal responses observed for these parameters were either equal to or greater than those previously reported for either isolated hepatocytes or perfused livers, demonstrating the sensitivity of this technique. Upon doseresponse examination of glucagon challenge, it was observed that high doses of glucagon (>16 nM) stimulate glucose production by activating the cAMP-second messenger cascade. In contrast, low doses (<4 nM) stimulate this process without production of intracellular cAMP or activation of cAMP-dependent protein kinase, suggesting the operation of cAMP-independent messenger. Since this system permits measurements of parameters common to many cellular processes, this methodology is suitable for addressing both pharmacological and toxicological questions.

Original languageEnglish (US)
Pages (from-to)339-354
Number of pages16
JournalJournal of Pharmacological Methods
Volume19
Issue number4
DOIs
StatePublished - 1988

Fingerprint

Metabolism
Liver
Cyclic AMP
Glucagon
Hormones
Glucose
Cyclic AMP-Dependent Protein Kinases
Chemical activation
Physiological Phenomena
Glycogenolysis
Controlled Environment
Second Messenger Systems
Glycogen
Toxicology
Rats
Hepatocytes
Potassium
Maintenance
Pharmacology

Keywords

  • cAMP
  • Glucose metabolism
  • Liver slice
  • Peptide hormone glucagon
  • Perifusion
  • Precision-cut

ASJC Scopus subject areas

  • Pharmacology

Cite this

Perifused precision-cut liver slice system for the study of hormone-regulated hepatic glucose metabolism. / Mckee, Roberta L.; Hruby, Victor J; Trivedi, Devendra B.; Johnson, David G; Gandolfi, A Jay; Krumdieck, Carlos L.; Brendel, Klaus.

In: Journal of Pharmacological Methods, Vol. 19, No. 4, 1988, p. 339-354.

Research output: Contribution to journalArticle

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