Insulin regulation of lipoprotein lipase in cultured isolated rat adipocytes

Robert H. Eckel, Judith E. Prasad, Philip A. Kern, Stephen Marshall

Research output: Contribution to journalArticlepeer-review

41 Scopus citations

Abstract

The cellular regulation of adipose tissue lipoprotein lipase by insulin was investigated using cultured isolated rat adipocytes. Evidence for sustained cell viability over 3 days included stability of specific [125I]insulin binding and adipocyte number. Lipoprotein lipase was measured in three functional compartments: 1) enzyme activity secreted into the culture medium, 2) activity releasable from cell suspensions by heparin, and 3) activity extractable from cells (after maximal heparin release) in deoxycholate and detergent. One day after preparation, these activities stabilized and were 1.3 ± 0.2,1.4 ± 0.2, and 7.7 ± 0.9 neq/106 cells min, respectively (n = 24, mean ± SEM). Insulin, added the day after preparation, produced a dose-dependent (1-400 ng/ml) increase in lipoprotein lipase releasable from cells by heparin at 2, 4, and 24 h. Insulin also increased intracellular enzyme measured as deoxycholate-detergent-solubilized activity extracted from previously heparin-released cells. However, insulin-mediated increases in culture medium enzyme only occurred subsequent to cellular effects. All insulin-mediated effects were prevented by cycloheximide (1 µg/ml). Thus, insulin increased two cellular pools of adipocyte lipoprotein lipase in a dose-dependent manner, but had no direct effect on enzyme secretion. Overall, cultured isolated rat adipocytes appear to be a valuable system for the study of lipoprotein lipase regulation at the level of the adipocyte.

Original languageEnglish
Pages (from-to)1665-1671
Number of pages7
JournalEndocrinology
Volume114
Issue number5
DOIs
StatePublished - 1984

ASJC Scopus subject areas

  • Endocrinology

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