Cloning and immunocytochemical localization of a cyclic nucleotide-gated channel alpha-subunit to all cone photoreceptors in the mouse retina.
Cyclic nucleotide-gated channels (CNGC) are ligand-gated ion channels that open and close in response to changes in the intracellular concentration of the second messengers, 3;,5;-cyclic adenosine monophosphate and 3;,5;-cyclic guanosine monophosphate. Most notably, they transduce the chemical signal produced by the absorption of light in photoreceptors into a membrane potential change, which is then transmitted to the ascending visual pathway. CNGCs have also been implicated in the signal transduction of other neurons downstream of the photoreceptors, in particular the ON-bipolar cells, as well as in other areas of the central nervous system. We therefore undertook a search for additional cyclic nucleotide-gated channels expressed in the retina. Following a degenerate reverse transcription polymerase chain reaction approach to amplify low-copy number messages, a cDNA encoding a new splice variant of CNGC alpha-subunit was isolated from mouse retina and classified as mCNG3. An antiserum raised against the carboxy-terminal sequence identified the retinal cell type expressing mCNG3 as cone photoreceptors. Preembedding immunoelectron microscopy demonstrated its membrane localization in the outer segments, consistent with its role in phototransduction. Double-labeling experiments with cone-specific markers indicated that all cone photoreceptors in the murid retina use the same or a highly conserved cyclic nucleotide-gated channel. Therefore, defects in this channel would be predicted to severely impair photopic vision.
Hirano
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Max-Planck-Institut für Hirnforschung, D-60528 Frankfurt am Main, Germany. hirano@mpih-frankfurt.mpg.de
Hack
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Wässle
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Duvoisin
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D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D054815:Cyclic Nucleotide-Gated Cation Channels; D018076:DNA, Complementary / Q000235:genetics / Q000502:physiology; D007150:Immunohistochemistry; D007473:Ion Channels / Q000096:biosynthesis / Q000502:physiology* / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D012160:Retina / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D017949:Retinal Cone Photoreceptor Cells / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D017948:Retinal Rod Photoreceptor Cells / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012374:Rod Cell Outer Segment / Q000378:metabolism / Q000648:ultrastructure; D015290:Second Messenger Systems / Q000502:physiology; D013045:Species Specificity
D054815:Cyclic Nucleotide-Gated Cation Channels; D018076:DNA, Complementary; D007473:Ion Channels