Brefeldin A differently affects basal and prolactin-stimulated milk protein secretion in lactating rabbit mammary epithelial cells.
When lactating mammary epithelial cells were treated with prolactin in vitro, numerous small vesicles rapidly accumulated in the Golgi area, and secretion of milk proteins increased. The effects of brefeldin A on these intracellular events were investigated. As observed by electron microscopy, stacks of the median Golgi were not altered after incubation in the presence of 50 nM brefeldin A but were dissociated when the drug concentration was > or = 500 nM. Small vesicles did not accumulate in the Golgi area when mammary cells were incubated in medium containing both prolactin and brefeldin A, whatever the concentration of the latter. Immunofluorescence experiments showed that 50 nM brefeldin A did not modify the localization of the CTR 433 median Golgi protein, but it induced redistribution of trans-Golgi network-associated proteins such as TGN38, AP-1 adaptor and clathrin. These effects occurred in the presence of brefeldin A plus prolactin. Pulse-chase experiments showed that brefeldin A concentrations > or = 100 nM induced the intracellular accumulation of milk proteins, provoked the appearance of immature forms of caseins, and inhibited milk protein secretion. In contrast, concentrations of brefeldin A of < or = 50 nM did not affect basal casein secretion but inhibited the secretagogue effect of prolactin. These data show not only that several biochemical events in the transport of milk proteins which are sensitive to different brefeldin A concentrations occur in lactating mammary epithelial cells, but also that it is possible to inhibit a hormonal stimulus in a selective manner, while the machinery responsible for basal secretion is still active.
Pauloin
A
A
INRA, Centre de Recherche de Jouy, France.
Delpal
S
S
Chanat
E
E
Lavialle
F
F
Aubourg
A
A
Ollivier-Bousquet
M
M
D000818:Animals; D020126:Brefeldin A; D002364:Caseins / Q000096:biosynthesis*; D003517:Cyclopentanes / Q000494:pharmacology*; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006056:Golgi Apparatus / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008321:Mammary Glands, Animal / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008565:Membrane Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D011388:Prolactin / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar
D002364:Caseins; D003517:Cyclopentanes; D008565:Membrane Proteins; D011500:Protein Synthesis Inhibitors; D020126:Brefeldin A; D011388:Prolactin