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B. Burnside, J. Lin-Jones, M. Wu, E. Parker; Expression of a Dominant-negative Acting Kinesin II Transgene in Xenopus Laevis Rod Photoreceptors Results in Rod Degeneration . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2821.
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Purpose: To investigate the effects of kinesin II dominant-negative acting transgene expression in Xenopus rod photoreceptors. Kinesin II is a microtubule motor concentrated in photoreceptor connecting cilia, and has been shown to be required for axoneme assembly and maintenance in both motile and sensory cilia in other systems. Methods: To examine the role of kinesin II in photoreceptors, we produced transgenic Xenops laevis with photoreceptor-specific expression of a dominant-negative construct previously shown to disrupt kinesin II function in Xenopus cultured cells (Le Bot, N. et al., J. Cell Biol., 143:1559-1573, 1998). The stalk-tail region of the kinesin II KIF3B subunit (XKlp3) was fused to enhanced green fluorescent protein (eGFP), subcloned into a vector containing the rod opsin promoter, and used to generate transgenic Xenopus. Tadpole retinas were analyzed for transgenic fusion protein expression and localization. The rod:cone ratio was determined from retinal sections of both control and transgenic tadpoles to determine any deleterious effects of transgene expression on rods. Results: Localization of the kinesin II transgenic fusion protein product to the rod synapse, inner segment and connecting cilium was similar to the pattern of expression of endogenous kinesin II in rods. Tadpoles containing the kinesin II transgene expressed the fusion protein transiently. The loss of transgene expression in tadpoles containing the dominant-negative acting kinesin II transgene is reflected in a reduction in the numbers of rods compared to cones. Conclusions: Rod-specific expression of a dominant negative kinesin II subunit in transgenic Xenopus tadpoles results in a reduction in the number of rods in the retina. Our results using a dominant-negative acting kinesin II transgene are consistent with previous studies reporting that photoreceptor-specific knockout of a kinesin II subunit in mice resulted in rod degeneration (Marszalek, J.R. et al., Cell, 102:175-187, 2000), and confirm that photoreceptor survival requires the availability of a functional kinesin II motor. CR: None Support: NIH grants EY 03575 and EY 06859 and Foundation for Fighting Blindness.
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