SUMMARY Hypophysectomized mice were treated daily for 28 days with 12 i.u. prolactin, 10 μg luteinizing hormone (LH), 10 μg LH plus 12 i.u. prolactin, 50 μg testosterone propionate (TP), or 50 μg TP plus 12 i.u. prolactin. The yield of spermatogenesis was studied quantitatively from the counts of spermatogonia, preleptotene and pachytene spermatocytes and spermatids in the seminiferous tubules at stage VII of spermatogenesis. Prolactin administered alone caused a small, but significant, increase in the yield of spermatogenesis. Treatment with a mixture of LH, follicle-stimulating hormone and growth hormone in amounts 1·5 times higher than those reported as contaminants of prolactin had similar effects. Injections of LH or TP caused partial restoration of spermatogenesis. The yield of spermatogenesis was significantly higher in animals given LH plus prolactin than in the animals given LH alone. Prolactin, however, did not augment the effects of TP on spermatogenesis. It is concluded that prolactin acts on the Leydig cells of the testis to increase their responsiveness to LH. More androgen appears to be produced under the influence of LH when prolactin is also present.