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

• Published in: Biochimica et biophysica acta 2012-02, Vol.1820 (2), p.84-89
• Main Authors: Oliveira, P.F., Alves, M.G., Rato, L., Laurentino, S., Silva, J., Sá, R., Barros, A., Sousa, M., Carvalho, R.A., Cavaco, J.E., Socorro, S.
• Format: Article
• Language: English
• Subjects: Alanine - biosynthesis; Cells, Cultured; diabetes mellitus; Energy metabolism; gene expression; Gene Expression Regulation - drug effects; genes; germ cells; Glucose; Glucose - metabolism; Glucose Transporter Type 3 - genetics; Glucose Transporter Type 3 - metabolism; glucose transporters; Human Sertoli cell; Humans; Insulin; Insulin - deficiency; Insulin - pharmacology; L-Lactate Dehydrogenase - genetics; L-Lactate Dehydrogenase - metabolism; Lactate; lactate dehydrogenase; Lactic Acid - biosynthesis; Male; messenger RNA; metabolism; metabolites; Monocarboxylic Acid Transporters - genetics; Monocarboxylic Acid Transporters - metabolism; Muscle Proteins - genetics; Muscle Proteins - metabolism; nuclear magnetic resonance spectroscopy; Pyruvates - metabolism; reverse transcriptase polymerase chain reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; secretion; Sertoli cells; Sertoli Cells - enzymology; Sertoli Cells - metabolism
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2011.11.006

Sertoli 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. hSC-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. Insulin 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. This is the first report regarding the effect of insulin-deprivation on hSC metabolism. [Display omitted] ► 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.