July 2018
Volume 59, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2018
Selectively knocking down hexokinase 2 in rods leads to retinal degeneration
Author Affiliations & Notes
  • Rui Zhang
    Clinical Ophthalmology and Eye Health, The University of Sydney, Sydney, New South Wales, Australia
  • Weiyong Shen
    Clinical Ophthalmology and Eye Health, The University of Sydney, Sydney, New South Wales, Australia
  • So-Ra Lee
    Clinical Ophthalmology and Eye Health, The University of Sydney, Sydney, New South Wales, Australia
  • Michelle X. Yam
    Clinical Ophthalmology and Eye Health, The University of Sydney, Sydney, New South Wales, Australia
  • Ling Zhu
    Clinical Ophthalmology and Eye Health, The University of Sydney, Sydney, New South Wales, Australia
  • Ashish Easow Mathai
    Clinical Ophthalmology and Eye Health, The University of Sydney, Sydney, New South Wales, Australia
  • Ching-Kang Jason Chen
    Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, United States
  • Mark C Gillies
    Clinical Ophthalmology and Eye Health, The University of Sydney, Sydney, New South Wales, Australia
  • Footnotes
    Commercial Relationships   Rui Zhang, None; Weiyong Shen, None; So-Ra Lee, None; Michelle X. Yam, None; Ling Zhu, None; Ashish Easow Mathai, None; Ching-Kang Chen, None; Mark Gillies, None
  • Footnotes
    Support  This project was supported by grants from the Lowy Medical Research Institute, Australia National Health and Medical Research Council (NHMRC) and Ophthalmic Research Institute of Australia.
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 1003. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Rui Zhang, Weiyong Shen, So-Ra Lee, Michelle X. Yam, Ling Zhu, Ashish Easow Mathai, Ching-Kang Jason Chen, Mark C Gillies; Selectively knocking down hexokinase 2 in rods leads to retinal degeneration. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1003.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The retina is a metabolically active tissue and its energy demands are normally met through aerobic glycolysis. However, the cell types that carry out glycolysis in the mammalian retina are unclear. The aim of this project was to use a cell-specific approach to study the consequences of selectively knocking down hexokinase 2 (HK2), the first rate-limiting enzyme in the glycolytic pathway, in rod photoreceptor cells of the mouse retina.

Methods : We crossed rhodopsin (RHO)-Cre mice (Li et al. Genesis 2005) with Hk2 flox mice (HK2fl/fl) to create RHO-Cre:HK2+/fl and RHO-Cre:HK2fl/fl mice, with RHO-Cre mice crossed with Z-EG and Rosa-LacZ Cre reporter mice serving as controls. Retinal changes were studied by immunostaining and Western blots using antibodies against Hk2, lactate dehydrogenase A (LDH-A), pyruvate dehydrogenase E1a (PDH-E1a), recoverin, blue- and red-green opsin. Fluorescence-labelled peanut agglutinin (PNA) was also used to study changes in cone photoreceptor apical processes.

Results : The rod-specific gene targeting in RHO-Cre mice was evidenced by reporter gene expression in rod photoreceptors after crossing them with Cre-reporter mice. Hk2 was expressed in ganglion cells, the inner nuclear layer, the outer plexform layer and the photoreceptor inner segments in the normal retina. Western blot analysis indicated that HK2 was knocked down by ~37% in the retina of RHO-Cre mice crossed with Hk2-floxed mice at 8-10 weeks of age, with no significant difference between RHO-Cre:HK2+/fl and RHO-Cre:HK2fl/fl mice. Findings from Western blots were confirmed by immunostaining for Hk2. Selectively knocking down Hk2 in rods led to photoreceptor degeneration, as evidenced by the loss of PNA-stained photoreceptor apical processes and reduced expression of recoverin, blue- and red-green opsin. Hk2 knockdown also led to reduced expression of LDH-A, but with little effect on PDH-E1α expression.

Conclusions : Our data provide evidence that the activity of HK2 in rods is important for the maintenance of photoreceptor health.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×