IOVS Molecular Human Reproduction
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(Investigative Ophthalmology and Visual Science. 2002;43:1884-1890.)
© 2002 by The Association for Research in Vision and Ophthalmology, Inc.

Identification of Differentially Expressed Genes in Mouse Pax6 Heterozygous Lenses

Bharesh K. Chauhan1,2, Weiyan Zhang2,3, Kveta Cveklova1, Marc Kantorow3 and Ales Cvekl1

1 From the Departments of Ophthalmology and Visual Sciences and Molecular Genetics, Albert Einstein College of Medicine, Bronx, New York; and the 3 Department of Biology, West Virginia University, Morgantown, West Virginia.

PURPOSE. Pax6 is a critical regulator of the developing lens, other ocular tissues, central nervous system, and pancreas. Downstream targets of Pax6 are largely unknown. The present study was designed to identify differentially expressed genes in Pax6 heterozygous and normal mouse lenses.

METHODS. RNAs from 8-week-old normal and Pax6 heterozygous mouse lenses were analyzed by both RT-PCR differential display and a candidate-gene approach. The expression levels of identified genes were confirmed by semiquantitative RT-PCR.

RESULTS. Eight transcripts encoding phosphatase inhibitor Pip-1 protein, heat shock protein Hsp40, Purkinje cell protein Pcp4, an expressed sequence tag (EST; AA331381) originally reported in a brain-specific library, Pitx3, and CBP, were confirmed to be downregulated in the Pax6 heterozygous mouse lenses. {alpha}B- and ßA3/A1-crystallin transcripts exhibited decreased expression in Pax6 heterozygous lenses, whereas the expression levels of other crystallins were virtually unchanged.

CONCLUSIONS. The present data identify eight genes with expression levels that are decreased in Pax6 heterozygous lenses and provide evidence that four functional categories of transcripts—namely, small hsps ({alpha}B-crystallin and Hsp40), crystallins ({alpha}B- and ßA3/A1-crystallin), transcription factors (Pitx-3 and CBP), and components of signal transduction cascades (Pip-1) are under direct or indirect transcriptional control by Pax6.




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Copyright © 2002 by the Association for Research in Vision and Ophthalmology