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We have isolated and characterized a P-45017 alpha cDNA fragment from a rat testis library. The partial length rat P-45017 alpha cDNA (1Kb) has high overall nucleotide and deduced amino acid similarity with human and bovine P-45017 alpha cDNA's and contains the conserved tridecapeptide and heme regions, the termination codon and polyadenylation site. Using this rat testis cDNA probe we measured P-45017 alpha mRNA levels of rat Leydig cells from animals treated with hCG. Temporal studies with a low hCG dose showed an early increase in mRNA levels returning to control values at later times, while a higher desensitizing dose caused a marked reduction in the mRNA (24 h) and a small recovery at 48 h. Fetal rat Leydig cells maintained in the presence of LH treated with estradiol showed a 70% decrease in P-45017 alpha mRNA levels and testosterone production followed closely the changes in P-45017 alpha mRNA. These studies suggest that gonadotropin stimulation and desensitization of P-45017 alpha dependent enzymes in the adult rat testis as well as estradiol induced desensitization in fetal Leydig cells are related to levels P-45017 alpha mRNA.