Gene Expression

Using RT-PCR, a separate expression of the testicular genes was determined. In brief, the TRIzol reagent (Invitrogen, Life Technologies, Carlsbad, CA, USA) was used to extract total RNA from around 100 mg of testicular tissue. Quantitative Nanodrop RNA samples of 1.8 or more A260/A280 were used to synthesize DNA using a cDNA synthesis package (Fermentas, Waltham, MA, United States). To amplify cDNA, the SYBR Green master mix and the primers listed in Table 1 with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a household gene were added. Data on amplification were analyzed using 2^−ΔΔ methods (26).

Primer sequences.

STAR, steroidogenic acute regulatory protein; CYP11A1, cytochrome P450 cholesterol side-chain cleavage enzyme; 3B-HSD, hydroxy-delta-5-steroid dehydrogenase, 3 beta-and steroid delta-isomerase 7; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.