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Department of Human Physiology (J.L.T., D.W.W.), School of Medicine, University of California, Davis, California 95616; and Division of Life Sciences (G.S.), Lawrence Berkeley National Laboratory, University of California, Berkeley, California 94720
Address all correspondence and requests for reprints to: Judith L. Turgeon, Ph.D., Department of Human Physiology, University of California, Davis, California 95616. E-mail: jlturgeon{at}ucdavis.edu
The PR is critical for normal female reproductive function in mammals, including primates, and the PR-knockout mouse is an important model for establishing PR targets. For example, LH secretion is significantly altered both in vivo and in vitro in female PR-knockout mice, but to establish specific mechanisms affected by the absence of the PR in the mouse requires characterization of wild-type mouse cell biology. As steps toward this, the aims were to establish whether altered LH secretion in PR-knockout mice reflects altered mouse gonadotrope cell lineage during development secondary to PR deletion and to test the assumption that PR in wild-type mouse pituitaries has the same exclusive gonadotrope localization and E2 and progesterone regulation as in rat and monkey pituitaries. As an in vitro model, dispersed pituitary cells from 2-wk ovariectomized wild-type or PR-knockout mice were cultured ± E2 for 3 d. These cells were subjected to dual immunofluorescence staining for PR and LH, PRL, or GH. The proportion of LH-gonadotropes (8–9%) and somatotropes (26–29%) was not different for PR-knockout and wild-type cultures with or without E2. Lactotrope composition (41–42%) was the same in wild-type and PR-knockout, and E2 resulted in a similar and significant increase in the proportion (57–59%) for both mouse types. Nuclear PR immunoreactivity was absent in all PR-knockout pituitary cells. For wild-type, all LH gonadotropes showed nuclear PR immunoreactivity that was up-regulated by E2 (>10-fold increase). Progesterone exposure for 10 h but not 3 h led to a 40% decrease in PR immunoreactivity in LH-gonadotropes. Unexpectedly, PR immunoreactivity also localized to all lactotropes and was up-regulated by E2 and down-regulated by progesterone. In summary, the absence of PR has no effect on the proportion of LH gonadotropes, lactotropes, and somatotropes in ovariectomized PR-knockout mouse pituitary cultures. For ovariectomized wild-type mice, gonadotropes in the in vitro model contain PR that is up-regulated by E2, but the downregulation by progesterone is modest, compared with that previously reported for an in vitro rat model. In contrast to rats and monkeys, E2-dependent PR also is present in lactotropes of ovariectomized wild-type mice. These results underscore the risks in assuming identical cell biology between rats and mice.
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