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Endocrinology Vol. 141, No. 6 1942-1950
Copyright © 2000 by The Endocrine Society

ARTICLES

Agouti-Related Protein-Like Immunoreactivity: Characterization of Release from Hypothalamic Tissue and Presence in Serum1

Ji-Yao Li, Susan Finniss, Ying-Kui Yang, Qun Zeng, Song-Yi Qu, Greg Barsh2, Chris Dickinson and Ira Gantz

Departments of Surgery (J.-Y.L., Y.-K.Y., Q.Z., S.-Y.Q., I.G.) and Pediatrics (S.F., C.D.), University of Michigan, Ann Arbor, Michigan 48109-0682; and Howard Hughes Medical Institute and Departments of Pediatrics and Genetics (G.B.), Stanford University, Stanford, California 94305-5428

Address all correspondence and requests for reprints to: Ira Gantz, M.D., 6504 MSRB I, 1150 W. Medical Center Drive, Ann Arbor, Michigan 48109-0682. E-mail: IGantz{at}UMich.edu

A novel RIA was used to examine the release of agouti-related protein-like immunoreactivity (AGRP-LI) from perfused rat hypothalamic tissue slices and to characterize AGRP-LI in rat serum. A continuous low level basal AGRP-LI release was observed from hypothalami of rats fed ad libitum before the rats were killed. Basal AGRP-LI release was 3-fold greater in rats fasted 48 h. In fasted animals leptin dose-dependently suppressed basal AGRP-LI release. In fed animals no change in basal AGRP-LI release was detected in response to 10-6 M {alpha}-MSH, orexin B, melanin-concentrating hormone, or serotonin. HPLC analysis of AGRP-LI in rat serum identified a single peak that eluted in close proximity to synthetic AGRP (87–132) and mouse [Leu127Pro]AGRP and that was identical to the peak seen in hypothalamic and adrenal tissue extracts. The serum concentration of AGRP-LI in rats fed ad libitum was 0.865 ± 0.323 nmol/liter (mean ± SE). Food deprivation resulted in a slow, but statistically significant rise in serum immunoreactivity at 48 h [1.174 ± 0.118 nmol/liter (mean ± SE)]. Bilateral adrenalectomy did not change serum levels of AGRP-LI. These studies demonstrate that in the rat there are different levels of basal hypothalamic AGRP-LI release in fed and fasted states and that in the fasted rat this release can be profoundly suppressed by leptin. These studies also suggest that AGRP is present in the systemic circulation of rats.




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