Presence of insulin receptors in cultured glial C6 cells. Regulation by butyrate.
The presence of insulin receptor and its regulation by butyrate and other short-chain fatty acids was studied in C6 cells, a rat glioma cell line. Intact C6 cells bind 125I-insulin in a rapid, reversible and specific manner. Scatchard analysis of the binding data gives typical curvilinear plots with apparent affinities of approx. 6 nM and 70 nM for the low-affinity (approx. 90% of total) and high-affinity (approx. 10% of total) sites respectively. Incubation with butyrate results in a time- and dose-dependent decrease of insulin binding to C6 cells. A maximal effect was found with 2 mM-butyrate that decreased the receptor by 40-70% after 48 h. Butyrate decreased numbers of receptors of both classes, but did not significantly alter receptor affinity. Other short-chain fatty acids, as well as keto acids, had a similar effect, but with a lower potency. Cycloheximide caused an accumulation of insulin receptors at the cell surface, since insulin binding increased and receptor affinity did not change after incubation with the inhibitor. Simultaneous addition of butyrate and cycloheximide abolished the loss of receptors produced by the fatty acid. In cells preincubated with butyrate, cycloheximide also produced a large increase in receptor numbers, showing that in the absence of new receptor synthesis a large pool of receptors re-appears at the surface of butyrate-treated cells.
Montiel
F
F
Unidad de Endocrinologia Experimental, Instituto de Investigaciones Biomedicas, C.S.I.C., Madrid, Spain.
Ortiz-Caro
J
J
Villa
A
A
Pascual
A
A
Aranda
A
A
D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002460:Cell Line; D005232:Fatty Acids, Volatile / Q000494:pharmacology; D007700:Kinetics; D014176:Protein Biosynthesis; D051381:Rats; D011972:Receptor, Insulin / Q000032:analysis / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism*
D002087:Butyrates; D005232:Fatty Acids, Volatile; D020148:Butyric Acid; D011972:Receptor, Insulin