5 alpha-reductase type 2 and androgen receptor expression in gonadotropin releasing hormone GT1-1 cells

Gonadal steroids are potent modulators of gonadotropin releasing hormone (G nRH) secretion, and androgen binding sites and 5 alpha -reductase activity have been found in the immortalized GnRH secreting cell line GT1-1, suggest ing the existence of a direct androgenic control of GnRH dynamics. Two isof orms of the 5 alpha -reductase have been cloned with very different biochem ical/functional properties: 5 alpha -reductase type 1 (widely distributed i n the body) and 5 alpha -reductase type 2 (confined in androgen target stru ctures). We have analysed whether, in GT1-1, androgen binding sites are lin ked to 'classical' androgen receptor, and which 5 alpha -reductase isoform is active. Reverse transcriptase-polymerase chain reaction analysis showed that the mRNAs coding for androgen receptor and for the two 5 alpha -reduct ase isoforms are all expressed in GT1-1 cells. However, the 5 alpha -reduct ase enzymatic reaction showed a peak of activity at a narrow pH around 5.5, the optimum for the 5 alpha -reductase type 2. The affinity for testostero ne, of the enzyme present in GT1-1 cells, was very similar to that observed for the recombinant type 2 isozyme expressed in yeasts. The data indicate that GT1-1 cells (i) express a 'classical' androgen receptor and (ii) conta in the 5 alpha -reductase type 2 isoform, a specific marker of androgen-res ponsiveness.