December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Expression Of A C-terminal Truncated Rhodopsin On A Rhodopsin Null Background
Author Affiliations & Notes
  • ES Lee
    Molecular and Cell Biology University of California Berkeley CA
  • BB Burnside
    Molecular and Cell Biology University of California Berkeley CA
  • JG Flannery
    Vision Science and the Helen Wills Neuroscience Institute UC Berkeley CA
  • Footnotes
    Commercial Relationships   E.S. Lee, None; B.B. Burnside, None; J.G. Flannery, None. Grant Identification: NIH Grant EY11123, Foundation for Fighting Blindness
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 2691. doi:
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    • Get Citation

      ES Lee, BB Burnside, JG Flannery; Expression Of A C-terminal Truncated Rhodopsin On A Rhodopsin Null Background . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2691.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Previous studies in transgenic animals have shown that mutant opsin (s334ter) harboring a 15 aa C-terminal truncation is found in the outer segment even though the majority of these truncated opsins mis-sort to other photoreceptor membranes. This investigation continues studies of the transport mechanism of normal and truncated mutant rhodopsin in rod photoreceptors. Methods: CTA9 mice expressing s334 mutant rhodopsin were crossed to rhodopsin null mice to generate animals on different knockout backgrounds. Eyes from mice of genotype (s334 +/-,rho -/-) and (s334 +/-, rho +/-) were processed at P7-10 for histology and immunocytochemistry to examine outer segment formation and rhodopsin distribution. Littermate and age-matched animals (rho +/+, rho +/-, rho -/-) were used as controls. Opsin levels were quantified by RT-PCR and Western Blotting. Results: Mice expressing C-terminal truncated rhodopsin s334ter in the absence of wildtype rhodopsin failed to form outer segments at postnatal day eight (P8), although mutant opsin was detected in the distal tip of the connecting cilium. Restoration of one copy of full-length wildtype rhodopsin in s334 +/-, rho +/- animals was sufficient to restore membrane accumulation and outer segment disk formation, albeit to a lower level compared to rho +/- controls. Conclusion: These data argue against a model of photoreceptor disk morphogenesis in which a bulk-flow mechanism soley governs rhodopsin transport to the outer segment during development.

Keywords: 517 photoreceptors • 606 transgenics/knock-outs • 527 protein structure/function 
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