Localization of phospholamban in smooth muscle using immunogold electron microscopy

doi:10.1083/jcb.107.2.555

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Localization of phospholamban in smooth muscle using immunogold electron microscopy

DG Ferguson, EF Young, L Raeymaekers and EG Kranias
Department of Physiology and Biophysics, University of Cincinnati, College of Medicine, Ohio 45267-0576.

Phospholamban, the putative regulator of the Ca2+-ATPase in cardiac sarcoplasmic reticulum, was immunolocalized in canine visceral and vascular smooth muscle. Gently disrupted tissues were labeled with an affinity-purified phospholamban polyclonal antibody and indirect immunogold, using preembedding techniques. The sarcoplasmic reticulum of smooth muscle cells was specifically labeled with patches of immunogold distributed in a nonuniform fashion, while the sarcolemma did not appear to contain any phospholamban. The outer nuclear envelopes were also observed to be heavily labeled with the affinity- purified phospholamban polyclonal antibody. These findings suggest that phospholamban may play a role in the regulation of cytoplasmic and intranuclear calcium levels in smooth muscle cells.
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