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      Expression of Candidate Pro-GnRH Processing Enzymes in Rat Hypothalamus and an Immortalized Hypothalamic Neuronal Cell Line

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          Abstract

          Since gonadotropin-releasing hormone (GnRH, also referred to as LHRH) is a major hormone regulating mammalian reproduction, identification of the processing steps involved in the conversion of the pro-LHRH to LHRH is fundamental to our understanding of its physiology. Extracts from immortalized LHRH neurons (GT1) were used to isolate the pro-LHRH intermediate products and to identify the enzymes which may participate in these conversions. The GT1 cells contain and secrete a pro-LHRH species that elutes at approximately 10,000-12,000 molecular weight. The pro-LHRH is metabolized to various N- and C-terminally modified LHRH products and to gonadotropin-releasing hormone-associated peptide (GAP). Analyses of these intermediates suggests that, at least, four different enzymatic steps are involved in pro-LHRH processing. Northern blot analyses reveal that prohormone convertase 2 (PC2), carboxypeptidase E, glutaminyl cyclase, and peptidyl-glycine α-amidating monooxygenase are expressed in the GT1 cells and rat hypothalamus. PC2 immunoreactivity is localized to the perikarya and beaded axon-like processes of these cells. SDS-PAGE analyses indicate that PC2 is biosynthesized, processed and secreted by the immortalized LHRH neurons. Our results indicate that the GT1 cell line may serve as a useful model to study the regulation of pro-LHRH processing and that it may also represent an important tool for dissecting the molecular and cellular basis of mammalian reproduction.

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          Author and article information

          Journal
          NEN
          Neuroendocrinology
          10.1159/issn.0028-3835
          Neuroendocrinology
          S. Karger AG
          0028-3835
          1423-0194
          1995
          1995
          09 April 2008
          : 62
          : 2
          : 166-177
          Affiliations
          aHormone Action Workgroup, Laboratory of Cellular and Molecular Pharmacology, National Institute of Environmental Health Sciences, Research Triangle Park, N.C., USA; bDepartment of Anatomy, Albert Szent-Györgyi Medical University, Szeged, Hungary; cJ.A. deSève Laboratory of Biochemical Neuroendocrinology, Clinical Research Insitute of Montreal (affiliated with ΓUniversité de Montréal), Que., Canada; dDivision of Biological Psychiatry, Duke University Medical Center, Durham, N.C., USA
          Article
          127001 Neuroendocrinology 1995;62:166–177
          10.1159/000127001
          8584116
          © 1995 S. Karger AG, Basel

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          Page count
          Pages: 12
          Categories
          Regulation of Gonadotropin-Releasing Hormone

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