Gene expression profiling of individual hypothalamic nuclei from single animals using laser capture microdissection and microarrays.

Abstract:

:In order to identify novel genes involved in appetite and body weight regulation we have developed a microarray based method suitable for detecting small changes in gene expression in discrete groups of hypothalamic neurons. The method is based on a combination of stereological sampling, laser capture microdissection (LCM), PCR based amplification (SuperAmp), and one-color cDNA microarray analysis. To validate the method we assessed and compared fasting induced changes in mRNA levels of Neuropeptide Y (NPY) and proopiomelanocortin (POMC) in the hypothalamic arcuate nucleus (ARC) of diet-induced obese rats using cDNA microarrays, quantitative PCR and in situ hybridization. All methods revealed statistically significant fasting-induced changes in NPY and POMC expression. An additional 3480 differentially expressed probes (fold change >1.22, t-test p=0.05) were identified in the microarray analysis. Our findings demonstrate a consistent gene expression pattern across three different gene expression detection methods and strongly suggest that LCM coupled microarray analysis combined with SuperAmp can be used as a semi-quantitative mRNA profiling tool. Importantly, the sensitivity of the method greatly improves the usefulness of the microarray technology for gene expression profiling in non-homogeneous tissues such as the brain.

journal_name

J Neurosci Methods

authors

Paulsen SJ,Larsen LK,Jelsing J,Janssen U,Gerstmayer B,Vrang N

doi

10.1016/j.jneumeth.2008.09.024

subject

Has Abstract

pub_date

2009-02-15 00:00:00

pages

87-93

issue

1

eissn

0165-0270

issn

1872-678X

pii

S0165-0270(08)00577-3

journal_volume

177

pub_type

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