September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
A role for both kif7 and kif17 in early photoreceptor development
Author Affiliations & Notes
  • Tylor R Lewis
    Cell Biology, Neurobiology & Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Sean R Kundinger
    Ophthalmology, Medical College of Wisconsin, Medical College of Wisconsin, Wisconsin, United States
  • Amira L Pavlovich
    Cell Biology, Neurobiology & Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Jonathan R Bostrom
    Cell Biology, Neurobiology & Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Brian Link
    Cell Biology, Neurobiology & Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Joseph C Besharse
    Cell Biology, Neurobiology & Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Tylor Lewis, None; Sean Kundinger, None; Amira Pavlovich, None; Jonathan Bostrom, None; Brian Link, None; Joseph Besharse, None
  • Footnotes
    Support  NEI R01 EY03222, NEI P30 EY001931, T32 EY014537
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 558. doi:
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    • Get Citation

      Tylor R Lewis, Sean R Kundinger, Amira L Pavlovich, Jonathan R Bostrom, Brian Link, Joseph C Besharse; A role for both kif7 and kif17 in early photoreceptor development. Invest. Ophthalmol. Vis. Sci. 2016;57(12):558.

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

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Abstract

Purpose : Kif17, a kinesin-2 family member, exhibits “tipping”, whereby it accumulates at the distal tip of the axoneme in primary cilia. Kif7, a kinesin-4 family member, additionally localizes to cilia tips, where it controls axoneme structure. We have previously shown that morpholino knockdown of kif17 causes disruption of OS formation at 72hpf. We hypothesize that genetic loss of either kif7, kif17, or both will negatively affect early development of the photoreceptor outer segment (OS).

Methods : TALENs were used to generate an 11bp deletion allele of kif17, while the CRISPR-Cas9 system was utilized to generate both a 74bp deletion allele and a 32bp insertion allele in kif7. Plastic histology was used to quantify both size and number of OS in control retinas compared to kif7-/- retinas, kif17-/- retinas, and compound kif7-/-; kif17-/- retinas. A qPCR panel of each zebrafish opsin (opn1sw1, opn1sw2, opn1mw2, opn1lw1, rho) was used to monitor each photoreceptor sub-type: UV cone, blue cone, green cone, red cone, and rod respectively.

Results : Plastic histology revealed that loss of either kif7 or kif17 led to reduced OS number and size in the central, but not ventral, retina at 72hpf. This effect was similar to that observed previously for morpholino knockdown of kif17. Loss of both kif7 and kif17 did not have an additive effect on OS morphology at 72hpf compared to loss of kif17 alone. However, normal OS number and size recovered by 6dpf. qPCR reveals markedly decreased expression of opn1sw1 in mutants compared to control embryos.

Conclusions : There is a developmental delay in OS formation in either kif7 or kif17 mutant fish. Similar to kif17 morphants, there were defects in OS morphogenesis prior to 3dpf. However, this delay is resolved morphologically by 6dpf. While OS achieve normal size, opn1sw1 expression remains abnormal, indicating that normal OS size can be achieved despite aberrations in opsin expression, particularly in UV cones. Work on the homeostatic mechanisms that control OS size despite altered expression of their principal gene products is ongoing.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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