The possible role of microtubules in milk production was examined in mammary gland slices from lactating guinea pigs. Colchicine, 10(-5)-10(-4) M, depressed lactose secretion within 15 min, maintaining maximal inhibition over 2.5 h, accompanied by retention of lactose within the slices. In colchicine dose-response studies (2 h, 10(-8)-10(-5) M), secretion was depressed 22% by 10(-5) M, whereas tissue lactose increased with dose up to +25% at 10(-5) M. Lactose synthesis was inhibited 3-19% without correlation to colchicine concentration. In another study, incubation with 10(-5) M lumicolchicine yielded one-third less inhibition of secretion than 10(-5) M colchicine with no increase in tissue lactose. Both drugs depressed synthesis 31%. Lactose secretion showed a negative correlation with 10(-7)-10(-4) M vincristine yielding a maximal 66% inhibition at 10(-4) M, whereas tissue retention showed a linear increase with concentration up to 151% of control at 10(-4) M. Effects on synthesis were sporadic. These data suggest that microtubules have a role in facilitating the transport and/or secretion of lactose and perhaps other milk components.