Biochemical and ultrastructural evidence for the association of basic fibroblast growth factor with cardiac gap junctions.
Basic fibroblast growth factor (bFGF) is a ubiquitous and multifunctional polypeptide that is believed to have a role in tissue repair and to act as a morphogen in embryonic development. Here, we have used immunohistochemical and biochemical methods with antibodies directed against the amino-terminal domain of bFGF, designated IS2, which recognize native and denatured bFGF, to demonstrate that in addition to its known intracellular and extracellular localization in heart, bFGF is also associated with cardiomyocyte gap junctions. In tissue sections, IS2 labeled regions of intercalated discs, producing an immunofluorescence pattern virtually indistinguishable from that obtained with antibodies against the heart gap junction protein connexin-43. By electron microscopy, gap junctions but not other regions of plasma membrane were heavily immunolabeled with this antibody. By solid phase immunoassay, bFGF was found to be more concentrated in a fraction enriched in cardiac gap junctions than in whole sarcolemmal preparations. Finally, an 18-kDa protein was recognized by several different antibodies specific for bFGF on Western blots of heart subcellular fractions enriched in gap junctions. We suggest that bFGF-like peptides are either an integral part of, or exist in close association with, cardiac gap junctions and thus may play a role in modulating gap junctional intercellular communication.
St. Boniface General Hospital Research Centre, Winnipeg, Manitoba, Canada.
E
E
Kardami
R M
RM
Stoski
B W
BW
Doble
T
T
Yamamoto
E L
EL
Hertzberg
J I
JI
Nagy
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000378:metabolism* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D007365:Intercellular Junctions / Q000648:ultrastructure*; D008854:Microscopy, Electron; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism / Q000648:ultrastructure
D011994:Recombinant Proteins; D005346:Fibroblast Growth Factors