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Endocrinology Vol. 140, No. 6 2781-2789
Copyright © 1999 by The Endocrine Society

ARTICLES

Stage- and Region-Specific Expression of Estrogen Receptor {alpha} Isoforms during Ontogeny of the Pituitary Gland1

Catherine Pasqualini, Dominique Guivarc’h, Ysander v. Boxberg, Fatiha Nothias, 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. Catherine Pasqualini or Dr. Philippe Vernier, Institut Alfred Fessard, UPR2212, Centre National de la Recherche Scientifique, avenue de la Terrasse, 91198 Gif-sur-Yvette-Cedex, France. E-mail: Catherine.Pasqualini{at}iaf.cnrs-gif.fr

The expression time course of estrogen receptor {alpha} (ER{alpha}) was analyzed by RT-PCR in fetal and newborn rat pituitaries. In addition to the classical ER{alpha} messenger RNA (mRNA), three shorter transcripts were detected and subsequently cloned. Sequence analysis showed that they corresponded to ER{alpha} mRNAs lacking exon 3 (which encodes a zinc finger in the DNA-binding domain), exon 4 (which encodes the nuclear localization signal and part of the steroid-binding domain), or both exons 3 and 4. As analyzed by RT-PCR and ribonuclease protection assay, the respective expression levels of the different transcripts varied dramatically during pituitary development; short forms appeared 4 days before full-length ER{alpha} mRNA. On Western blots from rat pituitaries of different ages, an ER{alpha}-specific antiserum labeled four protein bands of the expected molecular weights, revealing that all four ER{alpha} mRNAs are translated in vivo. Immunocytochemistry, using the same antiserum, showed the ER{alpha} to be present first in the cytosol of intermediate lobe cells (around embryonic day 16). Only 5 days later, nuclear staining became detectable in the anterior lobe. We argue that the observed cytosolic staining will be essentially due to short ER{alpha} isoforms, which are indeed more abundantly expressed in the intermediate lobe. These data suggest that during pituitary development, the activity of the ER{alpha} might be specifically regulated by differential splicing of its primary transcript, resulting in a differential subcellular localization of the isoforms.




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