Pituitary-specific and hormonally regulated gene expression directed by the rat proopiomelanocortin promoter in transgenic mice.
All aspects of POMC biosynthesis exhibit tissue-specific regulation. The single copy gene is highly expressed in anterior lobe (AL) corticotrophs and intermediate lobe (IL) melanotrophs of the pituitary gland and in the arcuate nucleus of the hypothalamus. POMC gene transcription in corticotrophs is induced by hypothalamic CRH and vasopressin and inhibited by adrenal glucocorticoids, while in melanotrophs it is predominantly regulated by beta-adrenergic neural input and dopamine. To identify the rat POMC (rPOMC) gene sequences necessary and sufficient to target expression and hormonal regulation in corticotrophs and melanotrophs, we generated 13 transgenic mice carrying rPOMC fusion genes. The genes consisted of 706 or 480 basepairs of rPOMC 5' flanking sequences ligated to either the E. coli LacZ gene encoding beta-galactosidase or the K1 mutant of the SV40 large T-antigen gene. Overall, half of the transgenic lines had reporter gene expression in their AL and IL in a pattern indistinguishable from ACTH immunohistochemistry. In three of these lines, beta-galactosidase or K1 T-antigen was localized by double immunofluorescence exclusively to ACTH-positive corticotrophs and melanotrophs. Transcriptional regulation of the rPOMC-LacZ fusion gene in response to hormonal manipulation was quantified by a fluorescence assay for beta-galactosidase enzyme activity in pituitary extracts. There was a 15-fold increase in AL enzyme activity after adrenalectomy and a 3-fold increase in IL activity after haloperidol treatment. X-gal histochemistry of pituitaries from hormonally treated mice confirmed the cellular specificity of these effects.(ABSTRACT TRUNCATED AT 250 WORDS)
Hammer
G D
GD
Neuroscience Program Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts 02111.
Fairchild-Huntress
V
V
Low
M J
MJ
D000324:Adrenocorticotropic Hormone / Q000494:pharmacology; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics / Q000276:immunology; D000242:Cyclic AMP / Q000494:pharmacology; D004247:DNA / Q000737:chemistry*; D004298:Dopamine / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D007763:Lac Operon; D051379:Mice; D008822:Mice, Transgenic; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D011333:Pro-Opiomelanocortin / Q000235:genetics* / Q000276:immunology; D011401:Promoter Regions, Genetic; D014667:Vasopressins / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics
D000957:Antigens, Viral, Tumor; D014667:Vasopressins; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone; D004247:DNA; D000242:Cyclic AMP; D001616:beta-Galactosidase; D004298:Dopamine