DNA synthesis inhibition and reduced functional differentiation of midpregnant mouse mammary epithelia on collagen gels.
Mammary epithelial cells were examined for a link between DNA synthesis and subsequent synthesis and secretion of casein. Cells isolated from mice midway through pregnancy and cultured on collagen gels spread to form monolayers (spreading phase). Release of monolayer/gels to float in surrounding culture medium induces synthesis and secretion of casein (secretory phase). DNA synthesis was blocked during the spreading phase with cytosine arabinofuranoside (ARA C). Culture medium was assayed for casein by direct quantification of protein from SDS-PAGE fluorographs, and by immunoblotting. When induced to become secretory, cells exposed to ARA C during the spreading phase showed a marked reduction of secretion of casein as compared to control cultures (72% reduction). In contrast, cells exposed to ARA C during the secretory phase (after monolayer formation was complete) showed no significant reduction in secretion of casein. Measurement of intracellular casein in secretory phase cells showed that reduced secretion of casein by cultures blocked during the spreading phase occurs as a consequence of reduced levels of casein synthesized, and not because of an inability to secrete intracellular accumulations. The inhibitor effect was specific; there was no significant reduction in levels of total intracellular protein synthesis, and neither cell spreading nor monolayer formation was impaired by treatments. These data support the notion that DNA synthesis is a prerequisite to functional differentiation of midpregnant mouse mammary epithelia maintained on floating collagen gels.
Trumpbour
V
V
Biology Board of Studies, University of California, Santa Cruz 95064.
Rocha
V
V
D000818:Animals; D002364:Caseins / Q000096:biosynthesis; D002454:Cell Differentiation; D002478:Cells, Cultured; D003094:Collagen / Q000502:physiology; D003561:Cytarabine / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis*; D004847:Epithelial Cells; D005260:Female; D066298:In Vitro Techniques; D008321:Mammary Glands, Animal / Q000166:cytology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000502:physiology*
D002364:Caseins; D003561:Cytarabine; D003094:Collagen; D004247:DNA