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Fluorescence laser microdissection reveals a distinct pattern of gene activation in the mouse hippocampal region.


ABSTRACT: A histoanatomical context is imperative in an analysis of gene expression in a cell in a tissue to elucidate physiological function of the cell. In this study, we made technical advances in fluorescence laser microdissection (LMD) in combination with the absolute quantification of small amounts of mRNAs from a region of interest (ROI) in fluorescence-labeled tissue sections. We demonstrate that our fluorescence LMD-RTqPCR method has three orders of dynamic range, with the lower limit of ROI-size corresponding to a single cell. The absolute quantification of the expression levels of the immediate early genes in an ROI equivalent to a few hundred neurons in the hippocampus revealed that mice transferred from their home cage to a novel environment have distinct activation profiles in the hippocampal regions (CA1, CA3, and DG) and that the gene expression pattern in CA1, but not in the other regions, follows a power law distribution.

SUBMITTER: Yoshioka W 

PROVIDER: S-EPMC3491666 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Fluorescence laser microdissection reveals a distinct pattern of gene activation in the mouse hippocampal region.

Yoshioka Wataru W   Endo Nozomi N   Kurashige Akie A   Haijima Asahi A   Endo Toshihiro T   Shibata Toshiyuki T   Nishiyama Ryutaro R   Kakeyama Masaki M   Tohyama Chiharu C  

Scientific reports 20121107


A histoanatomical context is imperative in an analysis of gene expression in a cell in a tissue to elucidate physiological function of the cell. In this study, we made technical advances in fluorescence laser microdissection (LMD) in combination with the absolute quantification of small amounts of mRNAs from a region of interest (ROI) in fluorescence-labeled tissue sections. We demonstrate that our fluorescence LMD-RTqPCR method has three orders of dynamic range, with the lower limit of ROI-size  ...[more]

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