Expression of the α1F calcium channel subunit by photoreceptors in the rat retina

Catherine Morgans, Phil Gaughwin, Ryszard Maleszka

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Purpose: The CACNA1F gene encodes a voltage-gated calcium channel α1 subunit, α1F, which is expressed in the human retina. Mutations in this gene cause incomplete X-linked congenital stationary night blindness (CSNB2). The aim of this study was to obtain the sequence of the rat α1F cDNA and localize the encoded polypeptide in the rat retina. Methods: The full-length rat α1F sequence was compiled from sequencing of overlapping α1F PCR fragments amplified from rat retinal cDNA. Antiserum was raised against a human α1F peptide. It was found that the human α1F peptide used to generate the antiserum was conserved at only 11 out of 19 residues in the cloned rat sequence. Therefore, antibodies were affinity purified against either the human α1F peptide or the equivalent rat peptide and used for immunofluorescent staining of rat retina sections. Results: The rat α1F amino acid sequence was found to be 91% and 95% identical to the human and mouse α1F sequences, respectively. Antibodies affinity purified against the human α1F peptide stained both the outer nuclear layer (ONL) and outer plexiform layer of rat retina sections. In contrast, staining with antibodies affinity purified against the corresponding rat α1F peptide was restricted to the ONL. Conclusions: The rat α1F amino acid sequence is highly homologous to the human and mouse sequences. The immunohistochemical results indicate the existence of distinct α1F isoforms or α1F-like channels, which are differentially distributed in the cell bodies and synaptic terminals of photoreceptors in the rat retina.

Original languageEnglish (US)
Pages (from-to)202-209
Number of pages8
JournalMolecular Vision
Volume7
StatePublished - 2001
Externally publishedYes

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Calcium Channels
Retina
Peptides
Antibody Affinity
Immune Sera
Amino Acid Sequence
Complementary DNA
Staining and Labeling
Presynaptic Terminals
Genes
Protein Isoforms

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Expression of the α1F calcium channel subunit by photoreceptors in the rat retina. / Morgans, Catherine; Gaughwin, Phil; Maleszka, Ryszard.

In: Molecular Vision, Vol. 7, 2001, p. 202-209.

Research output: Contribution to journalArticle

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abstract = "Purpose: The CACNA1F gene encodes a voltage-gated calcium channel α1 subunit, α1F, which is expressed in the human retina. Mutations in this gene cause incomplete X-linked congenital stationary night blindness (CSNB2). The aim of this study was to obtain the sequence of the rat α1F cDNA and localize the encoded polypeptide in the rat retina. Methods: The full-length rat α1F sequence was compiled from sequencing of overlapping α1F PCR fragments amplified from rat retinal cDNA. Antiserum was raised against a human α1F peptide. It was found that the human α1F peptide used to generate the antiserum was conserved at only 11 out of 19 residues in the cloned rat sequence. Therefore, antibodies were affinity purified against either the human α1F peptide or the equivalent rat peptide and used for immunofluorescent staining of rat retina sections. Results: The rat α1F amino acid sequence was found to be 91{\%} and 95{\%} identical to the human and mouse α1F sequences, respectively. Antibodies affinity purified against the human α1F peptide stained both the outer nuclear layer (ONL) and outer plexiform layer of rat retina sections. In contrast, staining with antibodies affinity purified against the corresponding rat α1F peptide was restricted to the ONL. Conclusions: The rat α1F amino acid sequence is highly homologous to the human and mouse sequences. The immunohistochemical results indicate the existence of distinct α1F isoforms or α1F-like channels, which are differentially distributed in the cell bodies and synaptic terminals of photoreceptors in the rat retina.",
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