Guinea pig horizontal cells express GABA, the GABA-synthesizing enzyme GAD 65 , and the GABA vesicular transporter

Chenying Guo, Arlene A. Hirano, Salvatore Stella, Michaela Bitzer, Nicholas C. Brecha

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

γ-Aminobutyric acid (GABA) is likely expressed in horizontal cells of all species, although conflicting physiological findings have led to considerable controversy regarding its role as a transmitter in the outer retina. This study has evaluated key components of the GABA system in the outer retina of guinea pig, an emerging retinal model system. The presence of GABA, its ratelimiting synthetic enzyme glutamic acid decarboxylase (GAD 65 and GAD 67 isoforms), the plasma membrane GABA transporters (GAT-1 and GAT-3), and the vesicular GABA transporter (VGAT) was evaluated by using immunohistochemistry with well-characterized antibodies. The presence of GAD 65 mRNA was also evaluated by using laser capture microdissection and reverse transcriptase-polymerase chain reaction. Specific GABA, GAD 65 , and VGAT immunostaining was localized to horizontal cell bodies, as well as to their processes and tips in the outer plexiform layer. Furthermore, immunostaining of retinal whole mounts and acutely dissociated retinas showed GAD 65 and VGAT immunoreactivity in both A-type and B-type horizontal cells. However, these cells did not contain GAD 67 , GAT-1, or GAT-3 immunoreactivity. GAD 65 mRNA was detected in horizontal cells, and sequencing of the amplified GAD 65 fragment showed approximately 85% identity with other mammalian GAD 65 mRNAs. These studies demonstrate the presence of GABA, GAD 65 , and VGAT in horizontal cells of the guinea pig retina, and support the idea that GABA is synthesized from GAD 65 , taken up into synaptic vesicles by VGAT, and likely released by a vesicular mechanism from horizontal cells.

Original languageEnglish (US)
Pages (from-to)1647-1669
Number of pages23
JournalJournal of Comparative Neurology
Volume518
Issue number10
DOIs
StatePublished - May 15 2010

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gamma-Aminobutyric Acid
Guinea Pigs
Retina
Enzymes
Messenger RNA
GABA Plasma Membrane Transport Proteins
Laser Capture Microdissection
Aminobutyrates
Glutamate Decarboxylase
Membrane Transport Proteins
Synaptic Vesicles
Reverse Transcriptase Polymerase Chain Reaction
vesicular GABA transporter
Protein Isoforms
Immunohistochemistry
Cell Membrane
GAT
Antibodies

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

Guo, Chenying ; Hirano, Arlene A. ; Stella, Salvatore ; Bitzer, Michaela ; Brecha, Nicholas C. / Guinea pig horizontal cells express GABA, the GABA-synthesizing enzyme GAD 65 , and the GABA vesicular transporter In: Journal of Comparative Neurology. 2010 ; Vol. 518, No. 10. pp. 1647-1669.
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Guinea pig horizontal cells express GABA, the GABA-synthesizing enzyme GAD 65 , and the GABA vesicular transporter . / Guo, Chenying; Hirano, Arlene A.; Stella, Salvatore; Bitzer, Michaela; Brecha, Nicholas C.

In: Journal of Comparative Neurology, Vol. 518, No. 10, 15.05.2010, p. 1647-1669.

Research output: Contribution to journalArticle

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