Cell-to-cell communication and expression of gap junctional proteins in human diabetic and nondiabetic skin fibroblasts: effects of basic fibroblast growth factor.
Wound healing involves the interactions of many cell types, and is controlled in part by growth factors. Intercellular communication mediated by gap junctions is considered to play an important role in the coordination of cellular metabolism duringthe growth and development of tissues and organs. Basic fibroblast growth factor (bFGF), known to be important in wound healing, has been found to increase Cx43 expression and intercellular communication in endothelial cells and cardiac fibroblasts. It has been proposed that an increased coupling is necessary for the coordination of these cells in wound healing and angiogenesis, and that one of the actions of bFGF is to modulate intercellular communication. The aim of our study was to evaluate the effects of bFGF on gap junctional intercellular communication (GJIC) in vitro, and the presence of gap junctional proteins connexin (Cx) 26, Cx32, and Cx43 in fibroblasts of diabetic and nondiabetic individuals. Fibroblast cell lines (n = 10) were cultured for 3 d in serum-free media with or without bFGF (3 ng/mL). Cells were evaluated for the rate of GJIC by using laser cytometry, and for the presence of Cx26, Cx32, and Cx43 by immunohistochemical and Western analyses. All cell types communicated via contact-dependent mechanisms. The rate of GJIC was greater (p < 0.01) for diabetic than for nondiabetic fibroblasts (4.1 +/- 0.01 vs 3.3 +/- 0.01%/min). bFGF increased (p < 0.01) the rate of GJIC for diabetic (4.9 +/- 0.01 vs 4.1 +/- 0.01%) and nondiabetic (4.1 +/- 0.01 vs 3.3 +/- 0.01%) fibroblasts. Immunohistochemistry identified Cx26 in the cytoplasm, Cx32 was not detected, and Cx43 was present on the cellular borders in all cultures. Image analysis of immunofluorescent staining demonstrated that bFGF increased (p < 0.05) Cx43 expression in diabetic and nondiabetic fibroblasts. Western immunoblot analysis revealed bands at 43-46 kD that were similar in volume for diabetic and nondiabetic fibroblasts. Thus, gap junctions involving Cx43 and GJIC among fibroblasts appear to be targets for bFGF. Fibroblasts of diabetic individuals appear to have an increased rate of cell-cell coupling, correlating with a decreased rate of proliferation.
Abdullah
K M
KM
Department of Surgery, University of North Dakota, Fargo 58102, USA. kaymabd@aol.com
Luthra
G
G
Bilski
J J
JJ
Abdullah
S A
SA
Reynolds
L P
LP
Redmer
D A
DA
Grazul-Bilska
A T
AT
D000328:Adult; D015153:Blotting, Western; D002450:Cell Communication; D002478:Cells, Cultured; D000072259:Connexin 26; D018031:Connexin 43 / Q000032:analysis; D017630:Connexins / Q000032:analysis*; D003924:Diabetes Mellitus, Type 2 / Q000473:pathology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005347:Fibroblasts / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D017629:Gap Junctions / Q000737:chemistry* / Q000502:physiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D010777:Photochemistry; D000097002:Gap Junction beta-1 Protein
D000072259:Connexin 26; D018031:Connexin 43; D017630:Connexins; D016222:Fibroblast Growth Factor 2; D000097002:Gap Junction beta-1 Protein; C000607042:GJB2 protein, human