Analysis of Transcription Factor MRNAs in Identified Oxytocin and Vasopressin Magnocellular Neurons Isolated by Laser Capture Microdissection

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2013 Jul 30

PMID 23894472

Citations 6

Authors

Madison Humerick

Jeffrey Hanson

Jaime Rodriguez-Canales

Daniel Lubelski

Omar M Rashid

Yasmmyn D Salinas

Yijun Shi

Todd Ponzio

Raymond Fields

Michael R Emmert-Buck

Harold Gainer

Affiliations

Soon will be listed here.

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.

Citing Articles

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A RNA-Seq Analysis of the Rat Supraoptic Nucleus Transcriptome: Effects of Salt Loading on Gene Expression.

Johnson K, Hindmarch C, Salinas Y, Shi Y, Greenwood M, Hoe S PLoS One. 2015; 10(4):e0124523.

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Laser capture microdissection: Big data from small samples.

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