Expression of genes involved in lipid and fatty acid metabolism in ex vivo cultured mouse testes

ORESTI, G.M.; ISOLER ALACARAZ, J.; KLAMPACHAS, A.; SANTIAGO VALTIERRA, FLORENCIA X.; AVELDAÑO, M.I.; DEL MAZO, J.

Congreso; LIV Annual meeting of the Society of Biochemistry & Molecular Biology (SAIB); 2018

Spermatogenesis has been achieved in vitro using neonatal mouse testes maintained in a gas-liquid interphase culture system. In this setting, it is possible to manipulate lipid metabolism to know its role during the spermatogenic process, thereby gathering information potentially useful for ex vivo spermatogenesis technology. Here, the progression of spermatogenesis was followed in mouse explants at cytological and histological levels and the gene expression of some proteins involved in lipid metabolism were compared in vitro vs in vivo. Two PUFA elongases (Elovl2 and Elovl4), fatty acid 2-hydroxylase (Fa2h), two fatty acid binding proteins (Fabp3 and Fabp9) and a diacylglycerol acyltransferase (Dgat2) were examined by RT-qPCR. Testis explants from 6 days-old CD1 mice were cultured for 22 days. Primary spermatocytes (PS) appeared at around days 10-12, and the first round spermatids (RS) emerged after day 18 to become abundant at day 22. The whole process showed a delay compared with that in vivo. Interestingly, akin to findings in vivo, mRNA levels of Elovl4 were high in the pre-meiotic phase to decrease thereafter, while those of Elovl2 steadily increased from days 1 to 22. Fabp3 mRNA also increased with time in the explants, linked to interstitial cells differentiation. Maximal expression Fa2h, Fabp9, and Dgat2 occurred at day 22 in culture, associated with the increase in RS numbers. Our results suggest that finding influences that promote lipid metabolism (growth factors, hormones) will be a promissory way to optimize spermatogenesis in explants. Supported by the MCIyU, Spain [BFU2013?42164-R, BFU2017?87095-R to JdM], PGI-UNS [24/B272 to GMO].