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Here we demonstrate that mitochondrial continuity is robust and fusion is frequent in freshly isolated rat ventricular myocytes, manifesting both as rapid content mixing events between adjacent organelles and slower, often long-distance events. We show that mitochondrial fusion decreases dramatically in culture because of the decay in contractile activity and, more specifically, the underlying calcium oscillations, which involve mitofusin 1 (Mfn1) abundance. 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