Transgenic mice expressing Cre-recombinase specifically in retinal rod bipolar neurons

Invest Ophthalmol Vis Sci. 2005 Oct;46(10):3515-20. doi: 10.1167/iovs.04-1201.

Abstract

Purpose: To establish a transgenic mouse line that expresses Cre-recombinase in retinal rod bipolar cells for the generation of rod bipolar cell-specific knockout mutants.

Methods: The IRES-Cre-cDNA fragment was inserted into a 173-kb bacterial artificial chromosome (BAC) carrying the intact Pcp2 gene, by using red-mediated recombineering. Transgenic mice were generated with the modified BAC and identified. The Cre-transgenic mice were crossed with ROSA26 and Z/EG reporter mice to detect Cre-recombinase activity.

Results: X-gal staining showed that strong Cre-recombinase activities were present in retinal inner nuclear layers and cerebellar Purkinje cells. Double staining with an anti-GFP antibody and an anti-PKCalpha antibody (specific for retinal rod bipolar cells) revealed that Cre-recombinase activity localized exclusively to the rod bipolar cells in the retina.

Conclusions: A mouse BAC-Pcp2-IRES-Cre transgenic line that expresses Cre-recombinase in retinal rod bipolar neurons has been established. Because mutations in some ubiquitously expressed genes may result in retinal degenerative diseases, the mouse strain BAC-Pcp2-IRES-Cre will be a useful new tool for investigating the effects of retinal rod bipolar cell-specific gene inactivation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Chromosomes, Artificial, Bacterial
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Galactosides / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Guanine Nucleotide Exchange Factors
  • Indoles / metabolism
  • Integrases / genetics
  • Integrases / metabolism*
  • Interneurons / enzymology*
  • Male
  • Mice
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Neuropeptides / genetics
  • Pregnancy
  • Purkinje Cells / enzymology
  • Retinal Rod Photoreceptor Cells / cytology*
  • Retinal Rod Photoreceptor Cells / embryology
  • beta-Galactosidase / metabolism

Substances

  • Galactosides
  • Guanine Nucleotide Exchange Factors
  • Indoles
  • Neuropeptides
  • Pcp2 protein, mouse
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Cre recombinase
  • Integrases
  • beta-Galactosidase
  • 5-bromo-4-chloro-3-indolyl beta-galactoside