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Physiol. Genomics (August 19, 2003). doi:10.1152/physiolgenomics.00062.2003
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Submitted on April 10, 2003
Accepted on August 13, 2003

Array analysis of gene expression in Connexin43 null astrocytes

Dumitru A Iacobas1*, Marcia Urban-Maldonado1, Sanda Iacobas1, Eliana Scemes1, and David C Spray1

1 Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY, USA

* To whom correspondence should be addressed. E-mail: diacobas{at}aecom.yu.edu.

Connexin43 (Cx43) is the most abundant gap junction protein in brain, where it is found primarily between astrocytes. Although the morphology of astrocytes from Cx43 null (KO) mice is similar to that of wildtype (WT) astrocytes, KO astrocytes exhibit reduced growth rate in culture. In order to evaluate the impact of deletion of Cx43 on other genes, including those encoding cell cycle proteins, we used DNA arrays to determine expression patterns in cultured astrocytes from sibling Cx43 null and WT mice. RNA samples extracted from astrocytes cultured from wild type and Cx43 null neonatal mice were dye labeled and individually co-hybridized with a reference of labeled cDNAs pooled from a variety of tissues on eight gene arrays containing 8975 mouse DNA sequences. Normal variability in expression of each gene was evaluated and incorporated into "expression scores" in order to statistically compare expression levels between WT and KO samples. In Cx43 null astrocytes, 4.1% of the 4998 adequately quantifiable spots were found to have significantly (p < 0.05) decreased hybridization compared to controls and 9.4% of the spots showed significantly higher hybridization. The significantly different spots corresponded to RNAs encoding 252 known proteins, many not previously linked to gap junctions, including transcription factors, channels and transporters, cell growth and death signals, enzymes and cell adhesion molecules. These data indicate a surprisingly high degree of impact of deletion of Cx43 on other astrocyte genes, implying that gap junction gene expression alters numerous processes in addition to intercellular communication.




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