Identification of the thyrotropin-releasing hormone precursor, its processing products, and its coexpression with convertase 1 in primary cultures of hypothalamic neurons: anatomic distribution of PC1 and PC2

Endocrinology
Eduardo A NillniP McMillan

Abstract

The processing of pro-TRH, has been extensively studied in our laboratory using a corticotropic cell line, AtT20, transfected with the pro-TRH gene. We have also demonstrated that the convertases PC1 and PC2 process pro-TRH to cryptic peptides in vitro. However, although these processing pathways have been well characterized in vitro, little is known about the processing and subcellular distribution of pro-TRH and its derived peptides in hypothalamic neurons, an endogenous source of pro-TRH and PC enzymes. In this study we used multiple approaches to identify, both biochemically and anatomically, the presence and localization of pro-TRH (26 kDa) and its processing products. We also investigated the presence of PC1 and PC2 enzymes and the coexpression of pro-TRH and PC1 messenger RNAs. Identification of the TRH precursor was demonstrated by 1) Western blot analysis of cellular extracts, 2) immunoprecipitation of radiolabeled pro-TRH followed by analysis on acrylamide gel electrophoresis, 3) fluorescence immunocytochemistry, and 4) immunoelectron microscopy. The presence of the convertases PC1 and PC2 was determined by Western blot analysis of cellular extracts and fluorescence immunocytochemistry. The coexpression of pro-TRH wit...Continue Reading

Citations

Jun 24, 1997·Proceedings of the National Academy of Sciences of the United States of America·M FurutaD F Steiner
Sep 10, 2008·The Journal of Biological Chemistry·Amparo RomeroEduardo A Nillni
Apr 23, 2009·Endocrinology·Kristen R Vella, Anthony N Hollenberg
Dec 19, 2009·PloS One·Işin CakirEduardo A Nillni
Oct 9, 2012·Neuron·Paul H Taghert, Michael N Nitabach
Apr 27, 2017·Briefings in Functional Genomics·Dean C Semmens, Maurice R Elphick
Jan 28, 2015·International Journal of Molecular Sciences·Toshio Takahashi, Noriyo Takeda
Oct 26, 1999·Endocrine Reviews·E A Nillni, K A Sevarino
Aug 24, 2006·American Journal of Physiology. Endocrinology and Metabolism·Veronica Paez EspinosaTheodore C Friedman
Dec 1, 1999·Endocrinology·E A NillniO H Pescovitz
Sep 12, 2020·Journal of Physiology and Biochemistry·María Silvina LandaCarlos José Pirola
Aug 8, 1997·The Journal of Biological Chemistry·P SchanerE A Nillni
Oct 6, 2005·The Journal of Biological Chemistry·Lawrence R MulcahyEduardo A Nillni

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