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The effect of various hormones on the incorporation of [¹⁴C]acetate into the fatty acids of pregnant mouse mammary gland explants in organ culture was studied.

Of the hormones insulin (I), ovine prolactin (P), bovine growth hormone (GH) and cortisol (F) tested singly, only insulin stimulated fatty acid synthesis. There was synergism between cortisol or prolactin with insulin. The greatest stimulation in fatty acid synthesis occurred when explants were incubated in a medium containing either I + F + P or I + F + GH.

Analysis by radio-gas-liquid chromatography of the fatty acids synthesized by explants after ¹⁴C labelling, showed that the pattern of fatty acids formed in the presence of I + F was distinctly different from that produced in the presence of I + F + P or I + F + GH. In the presence of I + F, the pattern of fatty acids resembled that found in mouse adipose tissue, whilst with I + F + P or I + F + GH the pattern resembled that of mouse milk fat.

Synthesis of RNA was essential for the stimulation of fatty acid synthesis in explants incubated in medium containing I + F + P or I + F + GH. Results obtained when DNA synthesis was blocked with mitomycin C suggest that mitosis is important for the induction of milk-fatty acid synthesis.

Puromycin had no effect for up to 8 h on explants which had been previously cultured in medium containing I + F, I + F + P or I + F + GH. This suggests a slow turnover rate of the enzymes involved in the synthesis of milk fatty acids.