Distribution of the Na(+)-Ca2+ exchange protein in mammalian cardiac myocytes: an immunofluorescence and immunocolloidal gold-labeling study.
The present study reports on the location of the Na(+)-Ca2+ exchanger in cardiac sarcolemma with immunofluorescence and immunoelectron microscopy. Both polyclonal and monoclonal antibodies to the Na(+)-Ca2+ exchanger were used. The mAb was produced from a hybridoma cell line generated by the fusion of mouse myeloma NS-1 cells with spleen cells from a mouse repeatedly immunized with isolated reconstituted canine cardiac Na(+)-Ca2+ exchanger (Philipson, K. D. S. Longoni, and R. Ward. 1988. Biochim. Biophys. Acta. 945:298-306). The polyclonal antibody has been described previously and reacts with three proteins (70, 120, 160 kD) in cardiac sarcolemma associated with the Na(+)-Ca2+ exchanger (Nicoll, D. A., S. Longoni, and K. D. Philipson. 1990. Science (Wash. DC). 250:562-565). Both the monoclonal and the polyclonal antibodies appear to react with extracellular facing epitopes in the cardiac sarcolemma. Immunofluorescence studies showed labeling of the transverse tubular membrane and patchy labeling of the peripheral sarcolemma. The immunofluorescent labeling clearly delineates the highly interconnected T-tubular system of guinea pig myocytes. This localization of the exchanger to the sarcolemma, with an apparent high density in the transverse tubules, was also seen with immunoelectron microscopy. It is of great interest that the Na(+)-Ca2+ exchanger, as the main efflux route for Ca2+ in heart cells, would be abundantly located in sarcolemma closest to the release of Ca2+.
Frank
J S
JS
Department of Medicine, UCLA School of Medicine 90024-1760.
Mottino
G
G
Reid
D
D
Molday
R S
RS
Philipson
K D
KD
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D002118:Calcium / Q000378:metabolism*; D002352:Carrier Proteins / Q000032:analysis* / Q000276:immunology; D002469:Cell Separation; D000939:Epitopes; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D009206:Myocardium / Q000737:chemistry* / Q000166:cytology; D051381:Rats; D012508:Sarcolemma / Q000737:chemistry*; D012964:Sodium / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger; D014909:Wheat Germ Agglutinins
D000906:Antibodies; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D000939:Epitopes; D019831:Sodium-Calcium Exchanger; D014909:Wheat Germ Agglutinins; C047939:fluorescein isothiocyanate-wheat germ agglutinin; D012964:Sodium; D016650:Fluorescein-5-isothiocyanate; D002118:Calcium