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      Cloning and Expression of Two Related Connexins from the Perch Retina Define a Distinct Subgroup of the Connexin Family

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          Abstract

          We have cloned cDNAs for two closely related connexins (Cx), Cx35 and Cx34.7, from a perch retinal cDNA library. Sequencing of PCR products from genomic DNA revealed that both connexins have an intron 71 bp after the translation initiation site; in Cx35, the intron is 900 bp in length, whereas in Cx34.7 it is ∼20 kb. Southern blots of genomic DNA suggest that the two connexins represent independent single copy genes. In Northern blots, Cx35 and Cx34.7 transcripts were detected in retina and brain; Cx34.7 also showed a weak signal in smooth muscle (gut) RNA. Antibodies against Cx35 labeled a 30 kDa band on a Western blot of retinal membranes, and in histological sections, the pattern of antibody recognition was consistent with labeling of bipolar cells and unidentified processes in the inner plexiform and nerve fiber layers. When expressed in Xenopus oocytes, Cx35 and Cx34.7 formed homotypic gap junctions, but the junctional conductance between paired oocytes expressing Cx35 was 10-fold greater than that recorded for gap junctional channels formed by Cx34.7. The homotypic gap-junctional channels were closed in a voltage-dependent manner but with relatively weak voltage sensitivity. Heterotypic gap junctions formed by Cx35 and Cx34.7 displayed junctional conductances similar to those of Cx34.7 homotypic pairs and showed a slightly asymmetric current–voltage relationship; the side expressing Cx35 exhibited a higher sensitivity to transjunctional potentials. An analysis of the sequence and gene structure of the connexin family revealed that perch Cx35 and Cx34.7, skate Cx35, and mouse Cx36 constitute a novel γ subgroup.

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          Author and article information

          Journal
          J Neurosci
          J. Neurosci
          jneuro
          jneurosci
          J. Neurosci
          The Journal of Neuroscience
          Society for Neuroscience
          0270-6474
          1529-2401
          1 October 1998
          : 18
          : 19
          : 7625-7637
          Affiliations
          [ 1 ]Lions of Illinois Eye Research Institute, Department of Ophthalmology and Visual Sciences, University of Illinois College of Medicine, Chicago, Illinois 60612,
          [ 2 ]Unité de Neurovirologie et Régénération du Système Nerveux, Institut Pasteur, Paris, CEDEX 15, France, and
          [ 3 ]Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115
          Article
          PMC6793016 PMC6793016 6793016 2365
          10.1523/JNEUROSCI.18-19-07625.1998
          6793016
          9742134
          73fd8ada-6658-450c-a1df-367e312b11d2
          Copyright © 1998 Society for Neuroscience
          History
          : 15 April 1998
          : 6 July 1998
          : 14 July 1998
          Categories
          Article
          Custom metadata
          5.00

          cloning,oocytes,neurons,retinal connexins,intercellular communication,gap junctions,γ connexin subgroup,channels

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