Total internal reflectance fluorescence imaging of genetically engineered ryanodine receptor-targeted Ca²⁺ probes in rat ventricular myocytes.

ABSTRACT: The details of cardiac Ca²⁺ signaling within the dyadic junction remain unclear because of limitations in rapid spatial imaging techniques, and availability of Ca²⁺ probes localized to dyadic junctions. To critically monitor ryanodine receptors' (RyR2) Ca²⁺ nano-domains, we combined the use of genetically engineered RyR2-targeted pericam probes, (FKBP-YCaMP, K_d=150nM, or FKBP-GCaMP6, K_d=240nM) with rapid total internal reflectance fluorescence (TIRF) microscopy (resolution, ∼80nm). The punctate z-line patterns of FKBP,²-targeted probes overlapped those of RyR2 antibodies and sharply contrasted to the images of probes targeted to sarcoplasmic reticulum (SERCA2a/PLB), or cytosolic Fluo-4 images. FKBP-YCaMP signals were too sma

SUBMITTER: Pahlavan S 

PROVIDER: S-EPMC5599148 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature