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Endocrinology Vol. 139, No. 10 4213-4221
Copyright © 1998 by The Endocrine Society

ARTICLES

Alternative Splicing of the D2 Dopamine Receptor Messenger Ribonucleic Acid Is Modulated by Activated Sex Steroid Receptors in the MMQ Prolactin Cell Line1

Dominique Guivarc’h, Jean-Didier Vincent and Philippe Vernier

Institut Alfred Fessard, UPR2212, Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette Cedex, France

Address all correspondence and requests for reprints to: Dr. Philippe Vernier, Institut Alfred Fessard, UPR2212, Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette Cedex, France. E-mail: vernier{at}iaf.cnrs-gif.fr

The two isoforms of the D2 dopamine receptor are generated by alternative splicing of the exon 6 of the premessenger RNA (pre-mRNA), changing the length of the third cytoplasmic loop involved in the coupling to G proteins. In the MMQ PRL cell line, sex steroid hormones modulated the proportion of the two D2 receptor isoforms. Under controlled culture conditions, 17ß-estradiol (E2) strongly favored the production of the long isoform of D2 mRNA over the short one, whereas both isoforms were equally abundant when culture medium was hormone depleted. In the presence of progesterone (P), E2 action was inhibited, and equal amounts of each D2 receptor isoform were produced in the cells. Hormone treatments never modified either the total amount of D2 receptor mRNA and D2 receptor binding sites or D2 receptor-mediated inhibition of adenylyl cyclase. Specific antagonists demonstrated that the activity of each hormone depended on their nuclear receptors. Inhibitors of gene transcription or translation also showed that their activity required protein synthesis. The expression of the short D2 receptor isoform was never prominent, even at the single cell level. Analysis of the intron sequence flanking alternative exon 6 showed that only the upstream intron presented two sequence tracts known to be targets for splicing factors. Taken together, these results provide converging evidence for a physiologically relevant mechanism by which sex steroid receptors could regulate the expression of a splicing factor favoring the production of the long dopamine D2 receptor isoform.




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