Intracorneal positioning of the lens in Pax6-GAL4/VP16 transgenic mice

Venkatesh Govindarajan, Wilbur R. Harrison, Ningna Xiao, Dongcai Liang, Paul A. Overbeek

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Purpose: The purpose of this study was to establish a GAL4/VP16-based binary transactivation system that was active in the lens and corneal epithelium of transgenic mice. Methods: We generated transgenic mice with the transcriptional transactivator GAL4/VP16 driven by a modified Pax6 promoter that is active in lens and corneal epithelial cells. We also generated and tested UAS-lacZ reporter mice. Wild type and transgenic mice were analyzed by histological, in situ, and Southern hybridization techniques. Results: Five families (OVE1931, OVE1934, OVE1935, OVE1936, and OVE1937) that carry the Pax6-GAL4/VP16 transgene were generated. Unexpectedly, mice from three of the transgenic lines showed ocular abnormalities. In the family OVE1936, cataracts were seen in the heterozygous mice at the time of eyelid opening and homozygotes showed microphthalmia. Transgenic mice in families OVE1931 and OVE1937 appeared normal. Histological analysis of ocular sections of OVE1934, OVE1935, and OVE1936 homozygous transgenic mice showed intracorneal positioning of the lens. The corneal stromal cells were disorganized and there was no distinctive corneal endothelial layer. In situ hybridizations showed robust expression of the GALVP16 transgene in the lens and corneal epithelial cells of the OVE1934, OVE1935, and OVE1936, but not in OVE1931 or OVE1937 families. Bigenic embryos generated by mating the Pax6-GAL4/VP16 mice to the UAS-lacZ mice showed that the GAL4/VP16 transgenic protein is functional and can induce eye-specific expression of a UAS-lacZ reporter gene. Conclusions: Our data suggest that (1) expression the GAL4/VP16 transgene induces changes in gene expression in lens cells, (2) that developmentally important genes are affected, and (3) that bigenic phenotypes will need to be interpreted with caution.

Original languageEnglish
Pages (from-to)876-886
Number of pages11
JournalMolecular Vision
Volume11
StatePublished - Oct 26 2005

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Transgenic Mice
Lenses
Transgenes
In Situ Hybridization
Herpes Simplex Virus Protein Vmw65
Epithelial Cells
Eye Abnormalities
Microphthalmos
Corneal Epithelium
Lac Operon
Trans-Activators
Homozygote
Eyelids
Stromal Cells
Reporter Genes
Cataract
Transcriptional Activation
Embryonic Structures
Phenotype
Gene Expression

All Science Journal Classification (ASJC) codes

  • Ophthalmology

Cite this

Govindarajan, V., Harrison, W. R., Xiao, N., Liang, D., & Overbeek, P. A. (2005). Intracorneal positioning of the lens in Pax6-GAL4/VP16 transgenic mice. Molecular Vision, 11, 876-886.

Intracorneal positioning of the lens in Pax6-GAL4/VP16 transgenic mice. / Govindarajan, Venkatesh; Harrison, Wilbur R.; Xiao, Ningna; Liang, Dongcai; Overbeek, Paul A.

In: Molecular Vision, Vol. 11, 26.10.2005, p. 876-886.

Research output: Contribution to journalArticle

Govindarajan, V, Harrison, WR, Xiao, N, Liang, D & Overbeek, PA 2005, 'Intracorneal positioning of the lens in Pax6-GAL4/VP16 transgenic mice', Molecular Vision, vol. 11, pp. 876-886.
Govindarajan V, Harrison WR, Xiao N, Liang D, Overbeek PA. Intracorneal positioning of the lens in Pax6-GAL4/VP16 transgenic mice. Molecular Vision. 2005 Oct 26;11:876-886.
Govindarajan, Venkatesh ; Harrison, Wilbur R. ; Xiao, Ningna ; Liang, Dongcai ; Overbeek, Paul A. / Intracorneal positioning of the lens in Pax6-GAL4/VP16 transgenic mice. In: Molecular Vision. 2005 ; Vol. 11. pp. 876-886.
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