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PMID: 8860239 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation and characterization of the alpha-subunit of the rat rod photoreceptor cGMP-gated cation channel.

Journal of molecular neuroscience : MN ·Vol. 6 ·No. 4 ·1995-00-00 ·Pages 289-302

Barnstable CJ, Wei J-Y

Abstract

A combination of genomic and PCR clones has been used to derive the full-length coding sequence of the alpha-subunit of the rat rod photoreceptor cGMP-gated cation channel. The sequence encodes a protein of 683 amino acids with a predicted molecular weight of 79,221. The sequence shows extensive homology with other rod cGMP-gated channels and also with the rat olfactory cyclic nucleotide-gated cation channel. When the full-length sequence of the rat rod channel was expressed in Xenopus oocytes it gave a conductance that responded to cGMP with an EC(50) of 42 mu M. No response to 2 mM cAMP was detected. The conductance was decreased in the presence of increasing concentrations of calcium. Both monoclonal and polyclonal antibodies were generated against a C-terminal peptide of the rat rod channel. On immunoblots of adult rat retinal membranes the antibodies recognized a band of 71 kDa, suggesting that the rat channel may undergo proteolytic cleavage in the retina, as has previously been found for the bovine rod channel. Immunocytochemical labeling of adult rat retinal sections detected prominent labeling over the rod photoreceptor outer segments with both monoclonal and polyclonal antibodies.

MeSH Terms
Animals Antibodies, Monoclonal Cloning, Molecular Cyclic GMP/physiology Female Immunoblotting Ion Channel Gating/physiology Ion Channels/chemistry,immunology,physiology Microinjections Molecular Sequence Data Oocytes/physiology Patch-Clamp Techniques RNA Probes Rats Restriction Mapping Retinal Rod Photoreceptor Cells/chemistry,physiology Sequence Homology, Amino Acid Xenopus
Chemicals
Antibodies, Monoclonal Ion Channels RNA Probes Cyclic GMP
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Barnstable C J
Department of Ophthalmology and Visual Science, Yale University, School of Medicine, New Haven, CT 06520-8061, USA.
Wei J-Y
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Article Info
Journal
Journal of molecular neuroscience : MN
Abbr.
J Mol Neurosci
ISSN
0895-8696
DOI
10.1007/BF02736787
Published
1995-00-00
Pages
289-302
Language
English
Region
United States
NLM ID
9002991
Subset
IM
External Links
PubMed source DOI: 10.1007/BF02736787 View journal details
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