title: Analysis of transcription factor mRNAs in identified oxytocin and vasopressin magnocellular neurons isolated by laser capture microdissection.
abstract: The oxytocin (Oxt) and vasopressin (Avp) magnocellular neurons (MCNs) in the hypothalamus are the only neuronal phenotypes that are present in the supraoptic nucleus (SON), and are characterized by their robust and selective expression of either the Oxt or Avp genes. In this paper, we take advantage of the differential expression of these neuropeptide genes to identify and isolate these two individual phenotypes from the rat SON by laser capture microdissection (LCM), and to analyze the differential expression of several of their transcription factor mRNAs by qRT-PCR. We identify these neuronal phenotypes by stereotaxically injecting recombinant Adeno-Associated Viral (rAAV) vectors which contain cell-type specific Oxt or Avp promoters that drive expression of EGFP selectively in either the Oxt or Avp MCNs into the SON. The fluorescent MCNs are then dissected by LCM using a novel Cap Road Map protocol described in this paper, and the purified MCNs are extracted for their RNAs. qRT-PCR of these RNAs show that some transcription factors (RORA and c-jun) are differentially expressed in the Oxt and Avp MCNs.
authors.affiliation: Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Madison
authors.initials: M
authors.lastname: Humerick
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Hanson
authors.forename: Jaime
authors.initials: J
authors.lastname: Rodriguez-Canales
authors.forename: Daniel
authors.initials: D
authors.lastname: Lubelski
authors.forename: Omar M
authors.initials: OM
authors.lastname: Rashid
authors.forename: Yasmmyn D
authors.initials: YD
authors.lastname: Salinas
authors.forename: Yijun
authors.initials: Y
authors.lastname: Shi
authors.forename: Todd
authors.initials: T
authors.lastname: Ponzio
authors.forename: Raymond
authors.initials: R
authors.lastname: Fields
authors.forename: Michael R
authors.initials: MR
authors.lastname: Emmert-Buck
authors.forename: Harold
authors.initials: H
authors.lastname: Gainer
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D061346:Laser Capture Microdissection / Q000379:methods*; D008297:Male; D010121:Oxytocin / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014157:Transcription Factors / Q000235:genetics*; D014667:Vasopressins / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D014157:Transcription Factors; D014667:Vasopressins; D010121:Oxytocin
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