Abstract
Antibodies against the main proteins in hepatic gap junctions--connexin-26 and connexin-32--have been used in conjunction with high-resolution patch-clamp techniques to investigate whether a relation exists between connexin type and conductance of single gap-junctional channels. Two different cell lines, BRL cells, derived from rat liver, and FL cells a human amniotic cell line exhibited the same single-channel conductances in double whole-cell recordings, but reacted differently upon dialysis with antibodies. Preliminary results indicate that both cell lines express mainly connexin-43. Thus, in spite of the inhibitory action of anti-cx26 and anti-cx32 antibodies observed, the data question the reliability of these antibodies for the functional characterization of gap-junction proteins in electrophysiological experiments.
MeSH Terms
Animals
Antibodies/physiology
Cells, Cultured
Connexin 26
Connexins
Electric Conductivity
Humans
Intercellular Junctions/physiology
Membrane Proteins/immunology
Perfusion
Chemicals
Antibodies
Connexins
GJB2 protein, human
Membrane Proteins
Connexin 26
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Paschke D
Abt. Biophysik, Universität Stuttgart, Federal Republic of Germany.
Eckert R
Hülser D F
References (16)
16 references, click to expand
-
Single-channel electrophysiology: use of the patch clamp.
Methods Enzymol. 1983;103:147-76
PMID: 6321886
-
Reduced number of gap junctions in rat hepatocarcinomas detected by monoclonal antibody.
Carcinogenesis. 1986 Sep;7(9):1475-82
PMID: 3017599
-
A computer method for the acquisition and analysis of patch-clamp single-channel currents.
J Neurosci Methods. 1987 May;20(1):45-55
PMID: 2438521
-
Expression of gap junction channels in communication-incompetent cells after stable transfection with cDNA encoding connexin 32.
Proc Natl Acad Sci U S A. 1990 Feb;87(4):1328-31
PMID: 2154741
-
Connexin family of gap junction proteins.
J Membr Biol. 1990 Jul;116(3):187-94
PMID: 2167375
-
Properties of single gap junctional channels between isolated neonatal rat heart cells.
Am J Physiol. 1988 Oct;255(4 Pt 2):H770-82
PMID: 2459974
-
Single channel currents of homo- and heterologous gap junctions between cardiac fibroblasts and myocytes.
Pflugers Arch. 1989 May;414(1):95-8
PMID: 2471143
-
Comparative characterization of the 21-kD and 26-kD gap junction proteins in murine liver and cultured hepatocytes.
J Cell Biol. 1989 Mar;108(3):1039-51
PMID: 2537831
-
Antibodies to gap-junctional protein selectively disrupt junctional communication in the early amphibian embryo.
Nature. 1984 Sep 13-19;311(5982):127-31
PMID: 6088995
-
Topological analysis of the major protein in isolated intact rat liver gap junctions and gap junction-derived single membrane structures.
J Biol Chem. 1987 Jun 5;262(16):7751-63
PMID: 3034905
-
Intracellular perfusion.
J Neurosci Methods. 1981 Oct;4(3):201-10
PMID: 6272031
-
Reduction of gap junctional conductance by microinjection of antibodies against the 27-kDa liver gap junction polypeptide.
Proc Natl Acad Sci U S A. 1985 Apr;82(8):2412-6
PMID: 2986116
-
Rates of diffusional exchange between small cells and a measuring patch pipette.
Pflugers Arch. 1988 Feb;411(2):204-11
PMID: 2451806
-
Structure of the junction between communicating cells.
Nature. 1980 Feb 7;283(5747):545-9
PMID: 7354837
-
Single-channel currents of an intercellular junction.
Nature. 1985 Sep 26-Oct 2;317(6035):331-5
PMID: 2413362
-
Cell-to-cell channel conductance during loss of gap junctional coupling in pairs of pancreatic acinar and Chinese hamster ovary cells.
Pflugers Arch. 1988 Jul;412(1-2):54-65
PMID: 2845356