Expression of nuclear hormone receptors in the rat supraoptic nucleus.
To investigate the role of nuclear hormone receptors on neuropeptide gene expression in the hypothalamo-neurohypophyseal system (HNS) of the rat, a survey was made of members of the nuclear hormone receptor superfamily that are expressed in the supraoptic nucleus (SON). A polymerase chain reaction cloning strategy based on homologies in the DNA-binding domain of AGGTCA-binding factors was devised for the identification of receptors in microdissected SON tissue. Cloning of the amplified products led to the identification of five true receptors, thyroid hormone receptor-alpha (THR alpha), retinoic acid receptor-alpha, retinoic acid receptor-gamma, retinoid X receptor-alpha, and retinoid X receptor-gamma, as well as four orphan receptors, apolipoprotein AI regulatory protein (ARP-1), chicken ovalbumin upstream promoter transcription factor I (COUP-TF I), estrogen-related receptor 2, and testis receptor 4 (TR4). Dot-blot screening of amplified gene fragment analysis showed that THR alpha, ARP-1, TR4, and COUP-TF I were the most abundant factors expressed in the SON region, in the order THR alpha > ARP-1 > TR4 approximately COUP-TF I. THR alpha has previously been localized to HNS neurons. In situ hybridization analysis showed that ARP-1, COUP-TF I, and TR4 were not expressed in magnocellular neurons at appreciable levels, but rather in surrounding structures. Furthermore, in lactating female rats there were no significant differences in the composition of the nine identified nuclear hormone receptors in the SON region compared with control animals. From these experiments, it is concluded that there is a multitude of hypothalamically expressed nuclear hormone receptors, but that only THR alpha is expressed at relatively high abundance in HNS neurons. This indicates that the peptide-producing magnocellular neurons of the SON express a specific set of transcription factors of the nuclear hormone receptor family.
Lopes da Silva
S
S
Rudolf Magnus Institute for Neurosciences, Department of Medical Pharmacology, Utrecht University, The Netherlands.
Van Horssen
A M
AM
Chang
C
C
Burbach
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002645:Chickens; D003001:Cloning, Molecular; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005260:Female; D015723:Gene Library; D007774:Lactation; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D011956:Receptors, Cell Surface / Q000096:biosynthesis*; D018160:Receptors, Cytoplasmic and Nuclear / Q000096:biosynthesis*; D018168:Receptors, Retinoic Acid / Q000096:biosynthesis; D011988:Receptors, Thyroid Hormone / Q000096:biosynthesis; D000072482:Retinoic Acid Receptor alpha; D047488:Retinoid X Receptors; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity; D013495:Supraoptic Nucleus / Q000378:metabolism*; D013737:Testis / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis; D000097768:Retinoic Acid Receptor gamma
D017931:DNA Primers; D004268:DNA-Binding Proteins; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; D018168:Receptors, Retinoic Acid; D011988:Receptors, Thyroid Hormone; D000072482:Retinoic Acid Receptor alpha; D047488:Retinoid X Receptors; D014157:Transcription Factors; D000097768:Retinoic Acid Receptor gamma; C000607073:Rara protein, rat