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PMID: 10931976 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Functional expression of the murine connexin 36 gene coding for a neuron-specific gap junctional protein.

The Journal of membrane biology ·Vol. 176 ·No. 3 ·2000-08-01 ·Pages 249-62

Teubner B, Degen J, Söhl G, Güldenagel M, Bukauskas FF, Trexler EB, Verselis VK, De Zeeuw CI, Lee CG, Kozak CA, Petrasch-Parwez E, Dermietzel R, Willecke K

Abstract

The mouse connexin 36 (Cx36) gene was mapped on chromosome 2 and an identical transcriptional start site was determined in brain and retina on exon I. Rabbit polyclonal antibodies to the presumptive cytoplasmic loop of the Cx36 protein recognized in immunohistochemical analyses Cx36 expression in the retina, olfactory bulb, hippocampus, inferior olive and cerebellum. In olivary neurons strong punctate labeling at dendritic cell contacts and weaker labeling in the cytoplasm of dendrites were shown by immuno electron microscopy. After expression of mouse Cx36 cDNA in human HeLa cells, neurobiotin transfer was increased 1.8-fold and electrical conductance at least 15-fold compared to untransfected HeLa cells. No Lucifer Yellow transfer was detected in either untransfected or Cx36 transfected HeLa cells. Single Cx36 channels in transfected HeLa cells showed a unitary conductance of 14.3 + or - 0. 8 pS. The sensitivity of Cx36 channels to transjunctional voltage was low in both HeLa-Cx36 cells and Xenopus oocytes expressing mouse Cx36. No increased transfer of neurobiotin was detected in heterotypic gap junctions formed by Cx36 and 9 other connexins expressed in HeLa cells. Our results suggest that Cx36 channels function as electrical synapses for transmission of electrical and metabolic signals between neurons in the central nervous system.

MeSH Terms
Animals Antibody Specificity Arachidonic Acid/pharmacology Base Sequence Biotin/analogs & derivatives,metabolism Brain/anatomy & histology,metabolism Carbon Dioxide/pharmacology Chromosome Mapping Connexins/biosynthesis,genetics,immunology,physiology Crosses, Genetic Electrophysiology Eye Proteins/biosynthesis,genetics,physiology Fluorescent Dyes/metabolism Gap Junctions/chemistry Genes HeLa Cells Humans Ion Channel Gating/drug effects Isoquinolines/metabolism Membrane Potentials Mice Mice, Inbred C57BL Molecular Sequence Data Muridae Nerve Tissue Proteins/biosynthesis,genetics,immunology,physiology Neurons/metabolism,ultrastructure Oocytes Rabbits Recombinant Fusion Proteins/biosynthesis Regulatory Sequences, Nucleic Acid Retina/metabolism Transcription, Genetic Transfection Xenopus laevis
Chemicals
Connexins Eye Proteins Fluorescent Dyes Isoquinolines Nerve Tissue Proteins Recombinant Fusion Proteins connexin 36 neurobiotin Carbon Dioxide Arachidonic Acid Biotin lucifer yellow
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Teubner B
Institut für Genetik, Abteilung Molekulargenetik, Universität Bonn, Römerstr. 164, D-53117 Bonn, Germany.
Degen J
Söhl G
Güldenagel M
Bukauskas F F
Trexler E B
Verselis V K
De Zeeuw C I
Lee C G
Kozak C A
Petrasch-Parwez E
Dermietzel R
Willecke K
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38 references, click to expand
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Article Info
Journal
The Journal of membrane biology
Abbr.
J Membr Biol
ISSN
0022-2631
Published
2000-08-01
Pages
249-62
Language
English
Region
United States
NLM ID
0211301
PMCID
PMC3659790
Subset
IM
Grants
NINDS NIH HHS · R01 NS036706 · United States
NINDS NIH HHS · NS367060 · United States
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