Effects of hyperglycemia and protein kinase C on connexin43 expression in cultured rat retinal pigment epithelial cells.
Previous results demonstrated that the intercellular communication mediated by gap junctions in retinal pigment epithelial (RPE) cells from the healthy Long Evans (LE) rat strain is higher than that from the dystrophic Royal College of Surgeons (RCS) rat strain. We examined connexin (Cx) expression in both cell types. At the mRNA level, a qualitatively similar expression pattern was found whereby Cx26, Cx32, Cx36, Cx43, Cx45 and Cx46 were all expressed. At the protein level, only Cx43 and Cx46 were detected. Expression of both isoforms was higher in LE-RPE as compared to RCS-RPE by a factor of 1.25 and 2 respectively. Phosphorylation of Cx43 was increased upon activation of protein kinase C (PKC) by 1 microM phorbol 12-myristate 13-acetate (PMA). The phosphorylation status was not changed in hyperglycemic conditions, but this treatment strongly decreased total Cx43 levels to about 75 and 40% (in LE-RPE and RCS-RPE cells respectively) of the control level in LE-RPE cells. This decrease could be overcome by PKC downregulation. These results demonstrate that PKC activation and hyperglycemic conditions have different effects on Cx43 and that PKC is involved in the metabolic pathway induced by hyperglycemic conditions.
Malfait
M
M
Laboratory of Physiology, KULeuven, Campus Gasthuisberg O/N, Herestraat 49 B-3000 Leuven, Belgium.
Gomez
P
P
van Veen
T A
TA
Parys
J B
JB
De Smedt
H
H
Vereecke
J
J
Himpens
B
B
D000818:Animals; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D018031:Connexin 43 / Q000096:biosynthesis* / Q000235:genetics; D017630:Connexins / Q000096:biosynthesis / Q000235:genetics; D003459:Crystallins / Q000096:biosynthesis* / Q000235:genetics; D004790:Enzyme Induction; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000494:pharmacology*; D006943:Hyperglycemia / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D011493:Protein Kinase C / Q000494:pharmacology*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D011922:Rats, Mutant Strains; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D018031:Connexin 43; D017630:Connexins; D003459:Crystallins; D020033:Protein Isoforms; D012333:RNA, Messenger; C082917:GJA3 protein, human; D011493:Protein Kinase C; D005947:Glucose; D013755:Tetradecanoylphorbol Acetate