Anti-ryanodine receptor antibody binding sites in vascular and endocardial endothelium.
The ryanodine receptor (RyR) functions as the calcium release channel of the sarcoplasmic reticulum activated by electromechanical coupling in skeletal and cardiac muscles. In smooth muscle, inositol trisphosphate releases calcium from internal stores during pharmacomechanical coupling, but these cells also contain ryanodine-sensitive calcium stores. In this study, we establish the presence of anti-RyR antibody binding sites in vascular and endocardial endothelium. Both types of endothelia also contain messenger RNA, which hybridizes to a cardiac RyR isoform cDNA probe. Western blots of endothelial cell homogenates demonstrate the presence of a single, high molecular weight band of protein that corresponds to the cardiac RyR isoform. Confocal micrographs of endothelial cells labeled with a specific anti-RyR antibody reveal an intense fluorescent signal surrounding the nucleus and distributed in a nonhomogeneous pattern throughout the cytoplasm. This pattern of fluorescence is consistent with the electron microscopic distribution of the endoplasmic reticulum. The pattern of immunofluorescence seen with the anti-RyR antibody is distinctly different from that seen with the mitochondrial fluorophore rhodamine 123. Our findings suggest that the RyR plays a role in endothelial signaling.
Lesh
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RE
Department of Anesthesiology, University of Virginia Health Sciences Center, Charlottesville 22908.
Marks
A R
AR
Somlyo
A V
AV
Fleischer
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S
Somlyo
A P
AP
D000818:Animals; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D015152:Blotting, Northern; D015220:Calcium Channels / Q000378:metabolism*; D004699:Endocardium / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D015151:Immunoblotting; D008297:Male; D008854:Microscopy, Electron; D009124:Muscle Proteins / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000276:immunology / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel
D015220:Calcium Channels; D009124:Muscle Proteins; D011950:Receptors, Cholinergic; D019837:Ryanodine Receptor Calcium Release Channel