A 221-bp fragment of the mouse opsin promoter directs expression specifically to the rod photoreceptors of transgenic mice

Alexander B. Quiambao, Neal S. Peachey, Nancy J. Mangini, P. Röhlich, Joe G. Hollyfield, Muayyad R. Al-Ubaidi

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Mutations in the human rod opsin gene have been shown to segregate with autosomal dominant retinitis pigmentosa (ADRP) and photoreceptor degeneration in transgenic mice. While these degenerations are characterized by the primary degeneration of rods, cones eventually die as well. To determine whether this subsequent cone degeneration is the result of expression of mutant rod opsin in the cones, the retinal cell-type specificity of a 221-bp fragment of the mouse rod opsin promoter was evaluated. Two transgenic mouse lines generated by injecting a fusion gene comprised of a 221-bp fragment of the mouse rod opsin promoter and the simian virus 40 large tumor antigen gene (Tag) were examined. The expression of Tag causes photoreceptor cell degeneration in members of both transgenic lines. However, the two lines differed with respect to the level of Tag expression and the rate and extent of photoreceptor cell degeneration. Immunocytochemical localization of opsin and Tag in surviving photoreceptor cells was determined and the results were confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). Rod- and cone-mediated function was evaluated by electroretinography (ERG). In the higher Tag-expressing transgenic line only one row of nuclei remained in the outer nuclear layer at postnatal day (P) 150. While these nuclei showed no antigenicity for rod opsin or Tag, they did stain with an antibody that reacts with both rod and cone S-antigens (arrestins), indicating that these cells were surviving photoreceptor nuclei. Positive staining with peanut agglutinin, which uniquely decorates matrix domains surrounding cones in the normal retina, confirmed that the surviving photoreceptor nuclei were of cone origin. RT-PCR substantiated the results from immunostaining; amplification product was obtained using blue cone opsin transcripts but nor from either Tag or rod opsin transcripts. The second transgenic mouse line exhibited a much slower photoreceptor cell death that was associated with low levels of Tag transgene transcript. At P120, ~50% of photoreceptors remained and an ~45% reduction in the rod ERG a-wave was observed. Cone-mediated ERGs, however, were normal. The results demonstrate the rod-specific expression of Tag as directed by the 221-bp fragment of the mouse rod opsin promoter and suggest that the cone degeneration in ADRP or transgenic mice associated with mutations in the rod opsin gene is a secondary effect of rod degeneration.

Original languageEnglish
Pages (from-to)617-625
Number of pages9
JournalVisual Neuroscience
Volume14
Issue number4
Publication statusPublished - Jul 1997

Fingerprint

Opsins
Retinal Rod Photoreceptor Cells
Rod Opsins
Neoplasm Antigens
Transgenic Mice
Photoreceptor Cells
Genes
Electroretinography
Vertebrate Photoreceptor Cells
Retinitis Pigmentosa
Reverse Transcriptase Polymerase Chain Reaction
Cone Opsins
Arrestins
Peanut Agglutinin
Retinal Cone Photoreceptor Cells
Mutation
Simian virus 40
Gene Fusion
Transgenes
Retina

Keywords

  • Cone photoreceptor
  • Opsin promoter
  • Retinal degeneration
  • T-antigen
  • Transgenic mice

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Quiambao, A. B., Peachey, N. S., Mangini, N. J., Röhlich, P., Hollyfield, J. G., & Al-Ubaidi, M. R. (1997). A 221-bp fragment of the mouse opsin promoter directs expression specifically to the rod photoreceptors of transgenic mice. Visual Neuroscience, 14(4), 617-625.

A 221-bp fragment of the mouse opsin promoter directs expression specifically to the rod photoreceptors of transgenic mice. / Quiambao, Alexander B.; Peachey, Neal S.; Mangini, Nancy J.; Röhlich, P.; Hollyfield, Joe G.; Al-Ubaidi, Muayyad R.

In: Visual Neuroscience, Vol. 14, No. 4, 07.1997, p. 617-625.

Research output: Contribution to journalArticle

Quiambao, AB, Peachey, NS, Mangini, NJ, Röhlich, P, Hollyfield, JG & Al-Ubaidi, MR 1997, 'A 221-bp fragment of the mouse opsin promoter directs expression specifically to the rod photoreceptors of transgenic mice', Visual Neuroscience, vol. 14, no. 4, pp. 617-625.
Quiambao, Alexander B. ; Peachey, Neal S. ; Mangini, Nancy J. ; Röhlich, P. ; Hollyfield, Joe G. ; Al-Ubaidi, Muayyad R. / A 221-bp fragment of the mouse opsin promoter directs expression specifically to the rod photoreceptors of transgenic mice. In: Visual Neuroscience. 1997 ; Vol. 14, No. 4. pp. 617-625.
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