Expression of connexin36 in cone pedicles and OFF-cone bipolar cells of the mouse retina

Andreas Feigenspan, Ulrike Janssen-Bienhold, Sheriar Hormuzdi, Hannah Monyer, Joachim Degen, Goran Söhl, Klaus Willecke, Josef Ammermüller, Reto Weiler

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    Abstract

    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 ß-galactosidase (ß-gal). In the outer nuclear layer, ß-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.
    Original languageEnglish
    Pages (from-to)3325-3334
    Number of pages10
    JournalJournal of Neuroscience
    Volume24
    Issue number13
    DOIs
    Publication statusPublished - 31 Mar 2004

    Fingerprint

    Retina
    Potassium Channels
    Reverse Transcriptase Polymerase Chain Reaction
    connexin 36
    Cone Opsins
    Neurokinin-3 Receptors
    Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels
    Immunohistochemistry
    Galactosidases
    Genetically Modified Animals
    Injections
    Metabotropic Glutamate Receptors
    AMPA Receptors
    Electrophysiology
    Glutamate Receptors
    Dendrites
    Reverse Transcription
    Exons
    Technology
    Light

    Cite this

    Feigenspan, A., Janssen-Bienhold, U., Hormuzdi, S., Monyer, H., Degen, J., Söhl, G., ... Weiler, R. (2004). Expression of connexin36 in cone pedicles and OFF-cone bipolar cells of the mouse retina. Journal of Neuroscience, 24(13), 3325-3334. https://doi.org/10.1523/JNEUROSCI.5598-03.2004
    Feigenspan, Andreas ; Janssen-Bienhold, Ulrike ; Hormuzdi, Sheriar ; Monyer, Hannah ; Degen, Joachim ; Söhl, Goran ; Willecke, Klaus ; Ammermüller, Josef ; Weiler, Reto. / Expression of connexin36 in cone pedicles and OFF-cone bipolar cells of the mouse retina. In: Journal of Neuroscience. 2004 ; Vol. 24, No. 13. pp. 3325-3334.
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    abstract = "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 {\ss}-galactosidase ({\ss}-gal). In the outer nuclear layer, {\ss}-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.",
    author = "Andreas Feigenspan and Ulrike Janssen-Bienhold and Sheriar Hormuzdi and Hannah Monyer and Joachim Degen and Goran S{\"o}hl and Klaus Willecke and Josef Ammerm{\"u}ller and Reto Weiler",
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    Feigenspan, A, Janssen-Bienhold, U, Hormuzdi, S, Monyer, H, Degen, J, Söhl, G, Willecke, K, Ammermüller, J & Weiler, R 2004, 'Expression of connexin36 in cone pedicles and OFF-cone bipolar cells of the mouse retina', Journal of Neuroscience, vol. 24, no. 13, pp. 3325-3334. https://doi.org/10.1523/JNEUROSCI.5598-03.2004

    Expression of connexin36 in cone pedicles and OFF-cone bipolar cells of the mouse retina. / Feigenspan, Andreas; Janssen-Bienhold, Ulrike; Hormuzdi, Sheriar; Monyer, Hannah; Degen, Joachim; Söhl, Goran; Willecke, Klaus; Ammermüller, Josef; Weiler, Reto.

    In: Journal of Neuroscience, Vol. 24, No. 13, 31.03.2004, p. 3325-3334.

    Research output: Contribution to journalArticle

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    T1 - Expression of connexin36 in cone pedicles and OFF-cone bipolar cells of the mouse retina

    AU - Feigenspan, Andreas

    AU - Janssen-Bienhold, Ulrike

    AU - Hormuzdi, Sheriar

    AU - Monyer, Hannah

    AU - Degen, Joachim

    AU - Söhl, Goran

    AU - Willecke, Klaus

    AU - Ammermüller, Josef

    AU - Weiler, Reto

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    PY - 2004/3/31

    Y1 - 2004/3/31

    N2 - 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 ß-galactosidase (ß-gal). In the outer nuclear layer, ß-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.

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