Elucidation of the structural basis of the excitation and contraction coupling usiing phase-contrast electron cryotomography

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We extracted enzymatically-active triad membranes from rabbit's skeletal muscle and examined them using electron cryomicroscopy. A lot of proteoliposomes were successfully observed in the sample. Some of them were identified as sarcoplasmic reticulum (SR). We further labeled the dihydropyridine receptor (DHPR) in T-tubules using the anti-beta subunit antibody conjugated fluorescent dyes and gold colloids to detect the triad junction (TJ). Although we can find gold-labels near SRs, The region of TJ has not been found out yet. We will also reconsider the condition of membrane extraction to preserve TJ abundantly, and examine it again in the future.