Effects of mild heat shock on glycogenesis and its regulation by insulin in cultured fetal hepatocytes.
The effects of a mild heat shock were investigated using cultured 15-day-old fetal rat hepatocytes in which an acute glucocorticoid-dependent glycogenic response to insulin was present. After exposure from 15 min to 2 h at 42.5 degrees C, cell surface [125I]insulin binding progressively decreased down to 60% of the value shown in cells kept at 37 degrees C, due toa decrease in the apparent number of insulin binding sites with little change in insulin receptor affinity. In parallel cultures, protein labeling with [35S]methionine exhibited stimulated synthesis of specific proteins, in particular, 73-kDa Hsc (heat shock cognate) and 72-kDa Hsp (heat shock protein). When cells were returned to 37 degrees C after 2 h at 42.5 degrees C, cell surface insulin binding showed a two-third restoration within 3 h (insulin receptor half-life = 13 h), with similar concomitant return of Hsps72,73 synthesis to preinduction levels. The rate of [14C]glucose incorporation into glycogen measured at 37 degrees C after 1- to 2-h heat treatment revealed a striking yet transient increase in basal glycogenesis (up to 5-fold). At the same time, the glycogenesis stimulation by insulin was reduced (from 3.2 to 1.4-fold), whereas that induced by a glucose load was maintained. Induction of thermotolerance after a first heating was obtained for the heat shock-dependent events except for the enhanced basal glycogenesis. In insulin-unresponsive cells grown in the absence of glucocorticoids, heat shock decreased the glycogenic capacity without modifying the glucose load stimulation, supporting the hypothesis that insulin and thermal stimulation of glycogenesis share at least part of the same pathway. Inverse variations were observed between Hsps72,73 synthesis and both cell surface insulin receptor level and insulin glycogenic response in fetal hepatocytes experiencing heat stress.
Laboratoire de Biologie-Odontologie, Université Paris 7, Institut Biomédical des Cordeliers, France.
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Zachayus
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D000818:Animals; D002478:Cells, Cultured; D006360:Heat-Shock Proteins / Q000096:biosynthesis; D006358:Hot Temperature; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology*; D008112:Liver Glycogen / Q000096:biosynthesis*; D014176:Protein Biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011972:Receptor, Insulin / Q000378:metabolism
D006360:Heat-Shock Proteins; D007328:Insulin; D008112:Liver Glycogen; D011972:Receptor, Insulin