Gap junctions in cultured astrocytes: single-channel currents and characterization of channel-forming protein.
Currents from gap junction channels were recorded from pairs of astrocytes in primary culture using the double whole-cell recording technique. In weakly coupled pairs, single-channel events could be resolved without pharmacological uncoupling treatment. Under these conditions, unitary conductance was 56 +/- 7 pS, and except for multiples of this value, no other level of conductance was observed consistently. To characterize the type of junctional protein constituting astrocyte gap junction channels, immunological and biochemical experiments were carried out on the same material. Specific cDNA probes for three connexins identified in mammals (Cx26, Cx32, and Cx43) showed that only Cx43 mRNA was expressed in cultured astrocytes. The presence of Cx43 protein in cultured astrocytes was demonstrated by immunoblotting, immunofluorescence, and immunogold labeling using anti-peptide antibodies specific to Cx43. These results strongly suggest that gap junctions in astrocytes have a 50-60 pS unitary conductance associated with channels composed of Cx43 protein.
Giaume
C
C
INSERM U114 Collége de France, Paris.
Fromaget
C
C
el Aoumari
A
A
Cordier
J
J
Glowinski
J
J
Gros
D
D
D000818:Animals; D001253:Astrocytes / Q000737:chemistry / Q000648:ultrastructure*; D002478:Cells, Cultured; D017630:Connexins; D003342:Corpus Striatum / Q000196:embryology / Q000648:ultrastructure; D004247:DNA / Q000235:genetics; D004594:Electrophysiology; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007150:Immunohistochemistry; D007365:Intercellular Junctions / Q000502:physiology*; D015640:Ion Channel Gating / Q000502:physiology; D007473:Ion Channels / Q000502:physiology*; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D051379:Mice; D009693:Nucleic Acid Hybridization; D051381:Rats
D017630:Connexins; D007473:Ion Channels; D008565:Membrane Proteins; D004247:DNA