Regulation of cholesterol side-chain cleavage cytochrome P450 in mouse testis Leydig cell line I-10

We have characterized regulation of steroidogenesis in a mouse testis Leydig cell line, I-10. Progesterone secretion was increased in a time- and dose-dependent fashion by 8-Br-cAMP treatment. The amount of cholesterol side-chain cleavage enzyme (P450scc), the first and rate-limiting enzyme for the synthesis of steroids, as detected by immunoblotting, was also increased. The calcium ionophore A23187 decreased the amount of P450scc, but it did not interfere with cAMP stimulation of the accumulation of P450scc. This regulation of P450scc expression by cAMP and A23187 is at the post-transcriptional level because the amount of P450scc mRNA was not affected by either treatment. This result was further confirmed by direct measurement of transcription in the presence or absence of forskolin treatment. I-10, however, supported cAMP-dependent transcriptional activation of the exogenous gene, as shown by the increased expression of a reporter gene under the control of the -600 to -2,500 fragment of the P450scc gene. The ability for transcriptional activation of the exogenous but not endogenous P450scc gene makes I-10 a unique steroidogenic cell line.