Alteration of Cx43:Cx40 expression ratio in A7r5 cells.
Connexins (Cx) 40 and 43 are coexpressed by several cell types at ratios that vary as a function of development, aging, and disease. Because these connexins form heteromeric channels, changes in expression ratio might be expected to significantly alter the connexin composition of the gap junction channel population and, therefore, gap junction function. To examine this possibility, we stably transfected A7r5 cells, which naturally coexpress Cx43 and Cx40, with a vector encoding antisense Cx43. Cx43 mRNA continued to be expressed in the antisense transfected clones, although levels were inversely related to the number of copies of antisense DNA incorporated into the genome. Protein levels, quantified in the clones with the highest and lowest Cx43:Cx40 mRNA ratios, were not well predicted by the mRNA levels, although the trends predicted by the Cx43:Cx40 mRNA ratio were preserved. Electrical coupling did not differ significantly between clones, but the clone with elevated Cx43:Cx40 protein expression ratio and unchanged Cx43 banding pattern was significantly better dye coupled than the parental A7r5 cells. These results suggest that as the Cx43:Cx40 ratio increases, provided alterations of Cx43 banding pattern (phosphorylation) have not occurred, permeability to large molecules increases even though electrical coupling remains nearly constant.
Burt
J M
JM
Department of Physiology, University of Arizona, Tucson, Arizona 85724, USA. jburt@u.arizona.edu
Fletcher
A M
AM
Steele
T D
TD
Wu
Y
Y
Cottrell
G T
GT
Kurjiaka
D T
DT
D016375:Antisense Elements (Genetics) / Q000494:pharmacology; D002450:Cell Communication / Q000502:physiology; D002460:Cell Line; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism*; D017630:Connexins / Q000235:genetics / Q000378:metabolism*; D004594:Electrophysiology; D017629:Gap Junctions / Q000378:metabolism / Q000502:physiology; D007473:Ion Channels / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics; D000097003:Gap Junction alpha-5 Protein
D016375:Antisense Elements (Genetics); D018031:Connexin 43; D017630:Connexins; D007473:Ion Channels; D012333:RNA, Messenger; D000097003:Gap Junction alpha-5 Protein