|Dyer, C - UNIVERSITY OF MISSOURI|
|Keisler, D - UNIVERSITY OF MISSOURI|
Submitted to: Society for Neuroscience Abstracts and Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: November 11, 1995
Publication Date: N/A
Technical Abstract: Neuropeptide Y (NPY) has been shown to exert effects upon the hypothalamic- pituitary-gonadal axis, but the mechanisms through which this peptide acts remain unclear. We report here the development of an ovine cDNA to the putative NPY receptor Y3 (also called LCR1). Ovine hypothalamic RNA was used in RT-PCR with primers derived from the published bovine Y3 sequence to amplify a 578 base pair cDNA. Cloning and sequencing revealed a >90% homology to the corresponding bovine sequence. Radiolabeled riboprobes derived from this ovine cDNA were used in a ribonuclease protection assay (RPA) to detect Y3 mRNA in ovine anterior and posterior hypothalamic and pituitary RNA extracts. High levels of Y3 mRNA expression were consistently detected in the pituitary extracts; much lower levels were detected in the posterior hypothalamus (containing the arcuate nucleus). No expression was detected by RPA in the anterior hypothalamus (containing the pre-optic area). The RNA extracts were then subjected to RT-PCR and Southern hybridization. Expression of Y3 mRNA was detected in all samples, demonstrating that low levels of Y3 mRNA, undetectable by RPA, exist within the anterior hypothalamus. These data suggest that the Y3 receptor mRNA is only weakly expressed in the hypothalamus but strongly expressed in the pituitary. Work is now underway to determine if gonadotrophs within the pituitary express Y3 mRNA.