May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Expression of Gaba, Gad65 and Vgat in Guinea Pig Horizontal Cells
Author Affiliations & Notes
  • C. Guo
    Neurobiology, David Geffen School of Medicine, Univ of California Los Angeles, Los Angeles, California
  • A. A. Hirano
    Neurobiology, David Geffen School of Medicine, Univ of California Los Angeles, Los Angeles, California
  • S. L. Stella, Jr.
    Neurobiology, David Geffen School of Medicine, Univ of California Los Angeles, Los Angeles, California
  • M. Bitzer
    Neurobiology, David Geffen School of Medicine, Univ of California Los Angeles, Los Angeles, California
  • N. C. Brecha
    Neurobiology, David Geffen School of Medicine, Univ of California Los Angeles, Los Angeles, California
    Jules Stein Eye Institute, Los Angeles, California
  • Footnotes
    Commercial Relationships  C. Guo, None; A.A. Hirano, None; S.L. Stella, None; M. Bitzer, None; N.C. Brecha, None.
  • Footnotes
    Support  EY15573 and VA Senior Career Scientist funds.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5795. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      C. Guo, A. A. Hirano, S. L. Stella, Jr., M. Bitzer, N. C. Brecha; Expression of Gaba, Gad65 and Vgat in Guinea Pig Horizontal Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5795. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : To evaluate the expression of GABA, its rate-limiting synthetic enzymes GAD (glutamate acid decarboxylase) 65 and 67, the vesicular GABA transporter (VGAT), and GABA plasma membrane transporter-1 (GAT-1) and GAT-3 in guinea pig outer retina. These studies are focused on testing the hypothesis that GABA, GAD and VGAT are expressed in horizontal cells of the guinea pig retina.

Methods: : Immunohistochemistry with antibodies to GABA, GAD65, GAD67, VGAT, GAT-1 and GAT-3 were used to evaluate adult guinea pig retina in vertical sections and after acute dissociation. Retinal whole-mounts were evaluated with antibodies to GAD65 and the horizontal cell marker calbindin. Horizontal cells were collected using laser capture microdissection (LCM). Total RNA was extracted from LCM microdissected horizontal cells using a RNA Nanoprep kit (Stratagene, CA). Degenerate RT-PCR was performed to detect GAD65 mRNA in horizontal cells.

Results: : Horizontal cells contained GABA, GAD65 and VGAT immunoreactivity, and they did not contain GAD67, GAT-1 and GAT-3 immunoreactivity, indicating the presence of GABA synthesis in horizontal cells. GABA, GAD65 and VGAT immunoreactivity was distributed to all horizontal cell bodies, as well as their processes and tips. GAD65, and GAD67 immunostaining in enzymatically dissociated horizontal cells and in retinal whole-mounts showed that GAD65 immunoreactivity was present in both A-type and B-type horizontal cells. Degenerate RT-PCR using primers specific to GAD65 detected the presence of GAD65 mRNA in horizontal cells. Sequencing of the amplified GAD65 fragment (126 bp) showed an 87% identity of the GAD65 mRNAs between guinea pig and mouse horizontal cells.

Conclusions: : These studies provide evidence for GABA synthesis in horizontal cells of the guinea pig retina and support the idea that vesicular transmitter release machinery is present in the mammalian horizontal cells.

Keywords: horizontal cells • gene/expression • retina 
×
×

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.

×