EXPRESSION OF GENES INVOLVED IN LIPID AND FATTY ACID METABOLISM IN EX VIVO CULTURED MOUSE TESTES

ISOLER-ALCARAZ J; DEL MAZO J; KAMPLACHAS A; ORESTI GM ; AVELDAÑO MI

Paraná, Entre Ríos

Congreso; LIV Reunión Anual de la Sociedad. Argentina de Invesgtigación en Bioquímica y Biología Molecular; 2018

Comisión directiva de la SAIB

Recently, spermatogenesis has been achieved in vitro using neonatal mouse testes maintained in a gas-liquid interphase culture system. In this setting, it is conceivable to manipulate lipid metabolism to comprehend the role that lipids play during the spermatogenic process, thereby gathering information potentially useful for the expansion of the 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 qPCR. Testis explants were prepared from 6 days-old CD1 mice and 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 enough at day 22. The whole process showed some 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 6 to 22. Fabp3 mRNA also increased with time in the explants, linked to interstitial cell 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 (e.g., growth factors, hormones) will be a promissory way to optimize spermatogenesis in explants.