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

2012; Nature Portfolio; Volume: 2; Issue: 1 Linguagem: Inglês

10.1038/srep00783

2045-2322

Wataru Yoshioka, Nozomi Endo, Akie Kurashige, Asahi Haijima, Toshihiro Endo, Toshiyuki Shibata, Ryutaro Nishiyama, Masaki Kakeyama, Chiharu Tohyama,

Cell Image Analysis Techniques

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.