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PMID: 7537274 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Specific permeability and selective formation of gap junction channels in connexin-transfected HeLa cells.

The Journal of cell biology ·Vol. 129 ·No. 3 ·1995-05-00 ·Pages 805-17

Elfgang C, Eckert R, Lichtenberg-Fraté H, Butterweck A, Traub O, Klein RA, Hülser DF, Willecke K

Abstract

DNAs coding for seven murine connexins (Cx) (Cx26, Cx31, Cx32, Cx37, Cx40, Cx43, and Cx45) are functionally expressed in human HeLa cells that were deficient in gap junctional communication. We compare the permeabilities of gap junctions comprised of different connexins to iontophoretically injected tracer molecules. Our results show that Lucifer yellow can pass through all connexin channels analyzed. On the other hand, propidium iodide and ethidium bromide penetrate very poorly or not at all through Cx31 and Cx32 channels, respectively, but pass through channels of other connexins. 4,6 Diamidino-2-phenylindole (DAPI) dihydrochloride shows less transfer among Cx31 or Cx43 transfectants. Neurobiotin is weakly transferred among Cx31 transfectants. Total junctional conductance in Cx31 or Cx45 transfected cells is only about half as high as in other connexin transfectants analyzed and does not correlate exactly with any of the tracer permeabilities. Permeability through different connexin channels appears to be dependent on the molecular structure of each tracer, i.e. size, charge and possibly rigidity. This supports the hypothesis that different connexin channels show different permeabilities to second messenger molecules as well as metabolites and may fulfill in this way their specific role in growth control and differentiation of cell types. In addition, we have investigated the function of heterotypic gap junctions after co-cultivation of two different connexin transfectants, one of which had been prelabeled with fluorescent dextran beads. Analysis of Lucifer yellow transfer reveals that HeLa cells expressing Cx31 (beta-type connexin) do not communicate with any other connexin transfectant tested but only with themselves. Two other beta-type connexin transfectants, HeLa-Cx26 and -Cx32, do not transmit Lucifer yellow to any of the alpha-type connexins analyzed. Among alpha-type connexins, Cx40 does not communicate with Cx43. Thus, connexins differ in their ability to form functional heterotypic gap junctions among mammalian cells.

MeSH Terms
Animals Cell Communication/physiology Cell Membrane Permeability/physiology Connexin 26 Connexins/biosynthesis,genetics,physiology Electric Conductivity Fluorescent Antibody Technique Gap Junctions/physiology HeLa Cells Humans Immunoblotting Ion Channels/physiology Mice Molecular Probes RNA, Messenger/analysis Transfection
Chemicals
Connexins GJB2 protein, human Ion Channels Molecular Probes RNA, Messenger Connexin 26
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Elfgang C
Institut für Genetik, Abt. Molekulargenetik, Universität Bonn, Germany.
Eckert R
Lichtenberg-Fraté H
Butterweck A
Traub O
Klein R A
Hülser D F
Willecke K
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.129.3.805
Published
1995-05-00
Pages
805-17
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120441
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.129.3.805 PMC Full Text View journal details
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