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The role of Pax6 splice variants in embryonic development

, : The role of Pax6 splice variants in embryonic development. Society for Neuroscience Abstract Viewer & Itinerary Planner : Abstract No 564 1

The transcription factor Pax6 is involved in regulating the development of the central nervous system, affecting such diverse processes as cell-cell adhesion, proliferation, and cellular differentiation. Pax6 contains two DNA-binding domains; a paired domain and a paired-like homeodomain. Alternative splicing of the Pax6 mRNA has been well documented, and a number of splice variants have been described. Best understood is the insertion of a 42 base pair fragment (exon 5a) within the paired domain, which is known to alter the DNA-binding properties of Pax6. A second variant, in which exon 6 is absent, is thought to abolish all DNA-binding via the paired domain. Alternative promoter usage is also important in the regulation of Pax6 expression, with transcripts being initiated from three promoters, designated P0, P1 and P-alpha.We have studied variations in the ratio of Pax6 isoforms present in the developing murine eye, forebrain, hindbrain and spinal cord across a variety of developmental stages from embryonic day 10.5 (E10.5) to E18.5. We have used RNase protection assays (RPA) to detect Pax6 mRNA. Transcripts containing exon 5a were found in a wide range of tissues of different ages, and the ratio of Pax6:Pax6+5a appears lower than previously suggested. Plasmid constructs have been made to overexpress both Pax6 and Pax6+5a, under the control of the CMV promoter, in immortalised neuroblastoma and fibroblast cell lines. These in vitro overexpression studies show differences in cellular behaviour mediated by the differing activities of the two isoforms, thus lending some functional significance to the in vivo expression data.Taken together, these data imply that the various Pax6 transcripts may play distinct, important roles in the development of the central nervous system.


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