Cloning and immunocytochemical localization of a cyclic nucleotide–gated channel α-subunit to all cone photoreceptors in the mouse retina
✍ Scribed by Arlene A. Hirano; Iris Hack; Heinz Wässle; Robert M. Duvoisin
- Publisher
- John Wiley and Sons
- Year
- 2000
- Tongue
- English
- Weight
- 733 KB
- Volume
- 421
- Category
- Article
- ISSN
- 0021-9967
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✦ Synopsis
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, 3Ј,5Ј-cyclic adenosine monophosphate and 3Ј,5Ј-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 ␣-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.