Effect of insulin deprivation on metabolism and metabolism-associated gene transcript levels of in vitro cultured human Sertoli cells

BackgroundSertoli cells metabolize glucose producing lactate for developing germ cells. As insulin regulates glucose uptake and its disturbance/insensitivity is associated with diabetes mellitus, we aimed to determine the effect of insulin deprivation in human Sertoli cell (hSC) metabolism and metabolism-associated gene expression.MethodshSC-enriched primary cultures were maintained in the absence/presence of insulin and metabolite variations were determined by 1H-NMR. mRNA expression levels of glucose transporters (GLUT1, GLUT3), lactate dehydrogenase (LDHA) and monocarboxylate transporter (MCT4) were determined by RT-PCR.ResultsInsulin deprivation resulted in decreased lactate production and in decrease of glucose consumption that was completely reverted after 6 h. Cells of both groups consumed similar amounts of glucose. In insulin-deprived cells, transcript levels of genes associated to lactate metabolism (LDHA and MCT4) were decreased. Transcript levels of genes involved in glucose uptake exhibited a divergent variation: GLUT3 levels were decreased while GLUT1 levels increased.Insulin-deprived hSCs presented: 1) altered glucose consumption and lactate secretion; 2) altered expression of metabolism-associated genes involved in lactate production and export; 3) an adaptation of glucose uptake by modulating the expression of GLUT1 and GLUT3.General significanceThis is the first report regarding the effect of insulin-deprivation on hSC metabolism.

Graphical abstractFigure optionsDownload full-size imageDownload high-quality image (50 K)Download as PowerPoint slideHighlights► The first hours of insulin deprivation are critical in hSCs in vitro. ► Insulin deprivation affects glucose uptake and lactate production/export. ► Insulin-deprived hSCs present altered expression of metabolism-associated genes. ► GLUT1 and GLUT3 expression levels are modulated by insulin-deprivation in hSCs.