Effects of transglutaminase or phospholipase A2 inhibitors on down-regulation of prolactin receptors and stimulation of casein and DNA synthesis in mammary gland explants.
A number of compounds have been found to inhibit the internalization of alpha 2-macroglobulin and/or epidermal growth factor into fibroblasts. Using the same inhibitor we tried to block the down-regulation of prolactin receptors, supposed to mirror internalization of PRL receptors, in order to investigate the possible role of internalization in the mechanism of prolactin action. In rabbit mammary cells it appeared that bacitracin and ethylamine completely block prolactin receptor down-regulation, but dansylcadaverine, the most potent inhibitor of transglutaminase, was without effect. The blockage of phospholipase A2 by chlorpromazine or bromophenacyl bromide (BPB) was without effect on the down-regulation of prolactin receptor. Subcellular distribution of [125I]oPRL has been studied after incubation with or without these various inhibitors after fractionation of mammary gland homogenate on sucrose gradient. Any of these compounds was able to increase the labelling of prolactin receptors located in plasma membrane fractions, suggesting that the rate of internalization of prolactin was not modified. In addition, none of these compounds inhibited the stimulation of beta-casein and DNA synthesis by prolactin. These results suggest that both transglutaminase and phospholipase A2 are not involved in the mechanism of prolactin-induced down-regulation of prolactin receptors, although bacitracin and ethylamine are able to block this phenomenon probably by different mechanism. In all cases, inductions of mitogenesis and beta-casein synthesis by prolactin in the rabbit mammary cells were not modified by the various compounds utilized. We conclude that neither transglutaminase nor phospholipase A2 are involved in the internalization of prolactin-receptor complexes, although bacitracin, ethylamine and quinacrine are able to block the down regulation of prolactin receptors by other means.
D000217:Acyltransferases / Q000037:antagonists & inhibitors*; D000818:Animals; D001414:Bacitracin / Q000494:pharmacology; D002364:Caseins / Q000096:biosynthesis*; D004247:DNA / Q000096:biosynthesis*; D005021:Ethylamines / Q000494:pharmacology; D005260:Female; D008321:Mammary Glands, Animal / Q000378:metabolism*; D009924:Organ Culture Techniques; D010740:Phospholipases / Q000037:antagonists & inhibitors*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D011388:Prolactin / Q000378:metabolism / Q000494:pharmacology; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D011981:Receptors, Prolactin; D011503:Transglutaminases
D002364:Caseins; D005021:Ethylamines; D011956:Receptors, Cell Surface; D011981:Receptors, Prolactin; D001414:Bacitracin; D011388:Prolactin; D004247:DNA; D000217:Acyltransferases; D011503:Transglutaminases; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2