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Optimized amplification and single-cell analysis identify GnRH-mediated activation of Rap1b in primary rat gonadotropes.

Authors: Yuen, T  Choi, SG  Pincas, H  Waring, DW  Sealfon, SC  Turgeon, JL 
Citation: Yuen T, etal., Mol Cell Endocrinol. 2012 Mar 5;350(1):10-9. doi: 10.1016/j.mce.2011.11.017. Epub 2011 Nov 25.
Pubmed: (View Article at PubMed) PMID:22127306
DOI: Full-text: DOI:10.1016/j.mce.2011.11.017

Identifying the early gene program induced by GnRH would help understand how GnRH-activated signaling pathways modulate gonadotrope secretory response. We previously analyzed GnRH-induced early genes in LbetaT2 cells, however these lack GnRH self-potentiation, a physiological attribute of gonadotropes. To minimize cellular heterogeneity, rat primary pituitary cultures were enriched for gonadotropes by 40-60% using a sedimentation gradient. Given the limited number of gonadotropes, RNA was amplified prior to microarray analysis. Thirty-three genes were up-regulated 40 min after GnRH stimulation. Real-time PCR confirmed regulation of several transcripts including fosB, c-fos, egr-2 and rap1b, a small GTPase and member of the Ras family. GnRH stimulated rap1b gene expression in gonadotropes, measured by a sensitive single cell assay. Immunocytochemistry revealed increased Rap1 protein in GnRH-stimulated gonadotropes. These data establish rap1b as a novel gene rapidly induced by GnRH and a candidate to modulate gonadotropin secretion in rat gonadotropes.

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CRRD Object Information
CRRD ID: 10041018
Created: 2015-05-08
Species: All species
Last Modified: 2015-05-08
Status: ACTIVE



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