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The Journal of Neuroscience, March 31, 2004, 24(13):3325-3334; doi:10.1523/JNEUROSCI.5598-03.2004
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Behavioral/Systems/Cognitive
Expression of Connexin36 in Cone Pedicles and OFF-Cone Bipolar Cells of the Mouse Retina
Andreas Feigenspan,1
Ulrike Janssen-Bienhold,1
Sheriar Hormuzdi,2
Hannah Monyer,2
Joachim Degen,3
Goran Söhl,3
Klaus Willecke,3
Josef Ammermüller,1 and
Reto Weiler1
1Institute of Biology, University of Oldenburg, D-26111 Oldenburg, Germany, 2Department of Clinical Neurobiology, University Hospital of Neurology, D-69120 Heidelberg, Germany, and 3Institute for Molecular Genetics, University of Bonn, D-53117 Bonn, Germany
Transgenic technology, immunocytochemistry, electrophysiology, intracellular injection techniques, and reverse transcription PCR were combined to study the expression of neuronal connexin36 (Cx36) in the outer plexiform layer of the mouse retina. Transgenic animals expressed either a fusion protein of full-length Cx36 with enhanced green fluorescent protein (EGFP) attached at the C terminus or exon 2 of Cx36 was replaced by -galactosidase (-gal). In the outer nuclear layer,-gal-positive cell bodies, which were confined to the most distal region close to the outer limiting membrane, displayed immunoreactivity against S-cone opsin. Cx36–EGFP puncta colocalized with cone pedicles, which were visualized by intracellular injection. In reverse transcriptase PCR experiments, Cx36 mRNA was never detected in samples of rods harvested from the outer nuclear layer. These results strongly suggest expression of Cx36 in cones but not in rods. In vertical sections, Cx36 expression in the vitreal part of the outer plexiform layer was characterized by a patchy distribution. Immunocytochemistry with antibodies against the neurokinin-3 receptor and the potassium channel HCN4 (hyperpolarization-activated cyclic nucleotide-gated potassium channel) displayed clusters of the Cx36 label on the dendrites of OFF-cone bipolar cells. In horizontal sections, these clusters of Cx36 appeared as round or oval-shaped groups of individual puncta, and they were always aligned with the base of cone pedicles. Double-labeling experiments and single-cell reverse transcriptase PCR ruled out expression of Cx36 in horizontal cells and rod bipolar cells. At light microscopic resolution, we found close association of Cx36–EGFP with the AMPA-type glutamate receptor subunit GluR1 but not with GluR2–GluR4, the kainate receptor subunit GluR5, or the metabotropic glutamate receptor mGluR6.
Key words: gap junctions; connexin; retina; rods; cones; bipolar cells; glutamate receptors
Received Sep 26, 2003;
revised January 22, 2004;
accepted January 23, 2004.
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