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Research ArticleNew Research, Sensory and Motor Systems

The Role of the Voltage-Gated Potassium Channel Proteins Kv8.2 and Kv2.1 in Vision and Retinal Disease: Insights from the Study of Mouse Gene Knock-Out Mutations

Nathan S. Hart, Jessica K. Mountford, Valentina Voigt, Paula Fuller-Carter, Melanie Barth, Jeanne M. Nerbonne, David M. Hunt and Livia S. Carvalho
eNeuro 11 February 2019, 6 (1) ENEURO.0032-19.2019; DOI: https://doi.org/10.1523/ENEURO.0032-19.2019
Nathan S. Hart
1School of Biological Sciences, The University of Western Australia, Perth, Western Australia 6009, Australia
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Jessica K. Mountford
1School of Biological Sciences, The University of Western Australia, Perth, Western Australia 6009, Australia
2Centre for Ophthalmology and Vision Science, Lions Eye Institute, The University of Western Australia, Perth, Western Australia 6009, Australia
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Valentina Voigt
2Centre for Ophthalmology and Vision Science, Lions Eye Institute, The University of Western Australia, Perth, Western Australia 6009, Australia
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Paula Fuller-Carter
2Centre for Ophthalmology and Vision Science, Lions Eye Institute, The University of Western Australia, Perth, Western Australia 6009, Australia
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Melanie Barth
2Centre for Ophthalmology and Vision Science, Lions Eye Institute, The University of Western Australia, Perth, Western Australia 6009, Australia
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Jeanne M. Nerbonne
3Department of Developmental Biology, Washington University School of Medicine, St. Louis, MO 63110
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David M. Hunt
1School of Biological Sciences, The University of Western Australia, Perth, Western Australia 6009, Australia
2Centre for Ophthalmology and Vision Science, Lions Eye Institute, The University of Western Australia, Perth, Western Australia 6009, Australia
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Livia S. Carvalho
2Centre for Ophthalmology and Vision Science, Lions Eye Institute, The University of Western Australia, Perth, Western Australia 6009, Australia
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  • Figure 1.
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    Figure 1.

    A, Targeting and excision of exon 1 of the Kcnv2 gene. B, qRT-PCR determination of the expression of the Kcnv2 gene in WT, Kv8.2 KO heterozygous and homozygous mice. Note complete absence of Kcnv2 expression in Kv8.2 KO homozygous tissues. Error bars indicate SEM. hBactP, human beta actin promoter; neo, neomycin; FRT, flip-recombinase targets.

  • Figure 2.
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    Figure 2.

    Retinal images and analysis of retinal layers and cell death. A, OCT images at six months of age. B, H & E images of transverse sections of WT and Kv8.2 KO retinae at six months of age. Scale bar = 100 µm. C, Thickness of retinal layers at six months of age, expressed as a % of WT. Kv8.2 KO, black; Kv2.1 KO, white; double KO, hatched. GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; OLM, outer limiting membrane; IS, inner segments; OS, outer segments; RPE, retinal pigmented epithelium. Kv8.2 KO, black; Kv2.1 KO, white; double KO, hatched. D, Cone cell loss in WT and Kv8.2 KO retinae at six months of age. E, TUNEL-positive cells in the retinae of WT (gray) and Kv8.2 KO (white) mice at three ages. Error bars indicate SEM; * and ** denote significance at the 5% and 1% probability level, respectively.

  • Figure 3.
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    Figure 3.

    ERG traces for WT and KO mice. A, a- and b-waves arising from 1-ms flashes at six stimulus intensities of 1, 10, 100, 1000, 5000, and 10,000 mcd⋅s m−2. Dotted lines and double-headed arrows indicate the a- and b-wave amplitudes at 10,000 mcd⋅s m−2. Top panel also shows comparative positions of OPs arising from 1-ms flashes at a stimulus intensity of 10,000 mcd⋅s m−2. The position of the four OPs are indicated by single arrows. B, c-wave arising from 1-ms flashes at 10,000 mcd⋅s m−2. C, Amplitudes of OPs. Error bars indicate SEM; * and ** denote significance at the 5% and 1% probability level, respectively.

  • Figure 4.
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    Figure 4.

    Amplitudes and implicit times for a-wave. A, a-wave amplitudes. B, Contribution of a-wave to b-wave amplitude. C, Implicit times for a-wave. WT, solid black line; Kv8.2 KO, solid gray line; Kv2.1 KO, dashed black line; double KO, dashed gray line. Error bars represent ± SEM.

  • Figure 5.
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    Figure 5.

    Amplitudes and implicit times for b-wave. A, b-wave amplitudes. B, Peak values of the positive component of b-wave. C, Peak values of b-wave response at four stimulus intensities showing. D, Implicit times for b-wave. WT, solid black line; Kv8.2 KO, solid gray line; Kv2.1 KO, dashed black line; double KO, dashed gray line; ** and * denotes significance at 1% and 5% probability levels, respectively. Error bars represent SEM.

  • Figure 6.
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    Figure 6.

    Dual flash recoveries. A, b-wave recovery. B, a-wave recovery. An initial 5-ms probe flash of 25,000 mcd⋅s m−2 was followed by a second test flash of equal intensity and duration at increasing intervals from 200 to 6000 ms, with a 30-s interval between each probe-test pair. WT, solid black line; Kv8.2 KO, dashed black line; Kv2.1 KO, solid gray line; double KO, dashed gray line. Error bars represent SEM.

  • Figure 7.
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    Figure 7.

    Photopic response in mice light-adapted at a background luminance of 50 cd m−2 before exposure to trains of 1-ms flashes of 10,000 mcd⋅s m−2 intensity at (A) 1 Hz (10 cycles, 1 s between repeats, repeated 10 times) and (B) 20 Hz (60 cycles, 1 s between repeats, repeated 10 times). C, Amplitudes at 1- and 10-Hz frequencies. WT, gray; Kv8.2 KO, green; Kv2.1 KO, blue; double KO, red; ** denotes significance at 1% probability level. Error bars represent SEM.

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The Role of the Voltage-Gated Potassium Channel Proteins Kv8.2 and Kv2.1 in Vision and Retinal Disease: Insights from the Study of Mouse Gene Knock-Out Mutations
Nathan S. Hart, Jessica K. Mountford, Valentina Voigt, Paula Fuller-Carter, Melanie Barth, Jeanne M. Nerbonne, David M. Hunt, Livia S. Carvalho
eNeuro 11 February 2019, 6 (1) ENEURO.0032-19.2019; DOI: 10.1523/ENEURO.0032-19.2019

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The Role of the Voltage-Gated Potassium Channel Proteins Kv8.2 and Kv2.1 in Vision and Retinal Disease: Insights from the Study of Mouse Gene Knock-Out Mutations
Nathan S. Hart, Jessica K. Mountford, Valentina Voigt, Paula Fuller-Carter, Melanie Barth, Jeanne M. Nerbonne, David M. Hunt, Livia S. Carvalho
eNeuro 11 February 2019, 6 (1) ENEURO.0032-19.2019; DOI: 10.1523/ENEURO.0032-19.2019
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Keywords

  • cone dystrophy
  • electroretinogram
  • photoreceptors
  • potassium channels
  • retina
  • retinal degeneration

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