Localization of the α1F calcium channel subunit in the rat retina

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

Purpose. The molecular identity of the calcium channels that mediate glutamate release from photoreceptors is unknown. Mutations in the recently identified, retina-specific α1F calcium channel subunit cause incomplete X-linked congenital stationary night blindness (CSNB2), the phenotype of which is consistent with a defect in neurotransmission within the retina. The purpose of this study was to determine the cellular distribution of the α1F subunit in the retina. Methods. Antibodies were raised against a unique peptide from the human a α1F sequence. Rat retina sections were labeled with affinity-purified α1F antibodies and the immunofluorescence analyzed by confocal microscopy. The α1F staining was compared with that obtained with a pan-α1 antibody, used to reveal the distribution of known voltage-gated calcium channels in the retina. Some sections were double labeled for α1F and the photoreceptor synaptic ribbon marker, bassoon. Results. Staining of retina sections with anti-α1F resulted in strong punctate labeling in the outer plexiform layer (OPL) and weak punctate labeling in the inner plexiform layer (IPL), consistent with a synaptic localization. Staining was also observed in the outer nuclear layer. Within the OPL, α1F immunoreactivity was clustered in discrete, horseshoe-shaped patches, the shape and dimensions of which are characteristic of rod active zones. Similar structures were labeled with the pan-α1 antibody. Localization of α1F immunoreactivity to rod active zones was confirmed by double labeling for bassoon, a component of photoreceptor synaptic ribbons. Conclusions. The distribution of α1F immunoreactivity in the OPL suggests that calcium influx through α1F or α1F-like channels mediates glutamate release from rod photoreceptors.

Original languageEnglish (US)
Pages (from-to)2414-2418
Number of pages5
JournalInvestigative Ophthalmology and Visual Science
Volume42
Issue number10
StatePublished - 2001
Externally publishedYes

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Calcium Channels
Retina
Antibodies
Staining and Labeling
Glutamic Acid
Retinal Rod Photoreceptor Cells
Confocal Microscopy
Synaptic Transmission
Fluorescent Antibody Technique
Calcium
Phenotype
Peptides
Mutation

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Localization of the α1F calcium channel subunit in the rat retina. / Morgans, Catherine.

In: Investigative Ophthalmology and Visual Science, Vol. 42, No. 10, 2001, p. 2414-2418.

Research output: Contribution to journalArticle

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abstract = "Purpose. The molecular identity of the calcium channels that mediate glutamate release from photoreceptors is unknown. Mutations in the recently identified, retina-specific α1F calcium channel subunit cause incomplete X-linked congenital stationary night blindness (CSNB2), the phenotype of which is consistent with a defect in neurotransmission within the retina. The purpose of this study was to determine the cellular distribution of the α1F subunit in the retina. Methods. Antibodies were raised against a unique peptide from the human a α1F sequence. Rat retina sections were labeled with affinity-purified α1F antibodies and the immunofluorescence analyzed by confocal microscopy. The α1F staining was compared with that obtained with a pan-α1 antibody, used to reveal the distribution of known voltage-gated calcium channels in the retina. Some sections were double labeled for α1F and the photoreceptor synaptic ribbon marker, bassoon. Results. Staining of retina sections with anti-α1F resulted in strong punctate labeling in the outer plexiform layer (OPL) and weak punctate labeling in the inner plexiform layer (IPL), consistent with a synaptic localization. Staining was also observed in the outer nuclear layer. Within the OPL, α1F immunoreactivity was clustered in discrete, horseshoe-shaped patches, the shape and dimensions of which are characteristic of rod active zones. Similar structures were labeled with the pan-α1 antibody. Localization of α1F immunoreactivity to rod active zones was confirmed by double labeling for bassoon, a component of photoreceptor synaptic ribbons. Conclusions. The distribution of α1F immunoreactivity in the OPL suggests that calcium influx through α1F or α1F-like channels mediates glutamate release from rod photoreceptors.",
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