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Monitoring synaptogenesis in the developing mouse cerebellum with an original oligonucleotide microarray
Author(s) -
Takahashi Masaki,
Kondoh Yasumitsu,
Tashiro Hideo,
Koibuchi Noriyuki,
Kuroda Yoichiro,
Tashiro Tomoko
Publication year - 2005
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.20533
Subject(s) - synaptogenesis , dna microarray , microarray , biology , oligonucleotide , gene , gene expression , cerebellum , computational biology , genetics , neuroscience , microbiology and biotechnology
Abstract Use of DNA microarrays in neuroscience have been limited to rough screening purposes, mainly because the reliability and sensitivity of available arrays are not high enough. Because only a few hundred to one thousand genes are usually found to change expression levels in most experiments, we attempted to develop a more quantitative array by the following strategies: 1) limit the genes to those relevant to the experimental system, 2) design oligonucleotide probes of specified molecular properties so that more stringent hybridization conditions can be employed, 3) place six spots per probe on one slide and use multiple normalization genes, and 4) use a new type of gold‐coated slide with higher S/N ratio. Genes involved in the construction and functioning of the synapse were selected from the literature as well as experimentally by comparing cerebella from hypothyroid and control mice at postnatal day 15 (P15). Although the number of genes covered was not yet large (172 genes), the custom array “Synaptoarray” thus constructed was capable of detecting ±20% difference in gene expression ratios. Analysis of the postnatal development of the mouse cerebellum by using Synaptoarray demonstrated a general expression pattern with a peak at P7, followed by a decline at P15 and a partial recovery after P21. P10 clearly marked the end of the initial growth stage and a major transcriptional turning point in this sytem. This result suggests that such a custom array should be useful in monitoring perturbations to the normal developmental program. © 2005 Wiley‐Liss, Inc.