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Effects of repetitive tetanic stimulation at long intervals on excitation‐contraction coupling in frog skeletal muscle.

1. Single skeletal muscle fibres of Xenopus frogs were used to investigate the possibility that excitation‐contraction (E‐C) coupling can be impaired under conditions of elevated intracellular free Ca2+ ([Ca2+]i). 2. Fibres were stimulated with a train of up to 200 tetani at 10 or 20s intervals; this long‐interval stimulation (LIS) scheme was chosen to minimize fatigue. After LIS, fibres were exposed to hypotonic Ringer solution for 5 min. At the end of LIS, force was about 90% of the original and the hypotonic challenge did not result in any force depression. 3. Caffeine, terbutaline and 2,5‐di(tert‐butyl)‐1,4‐benzohydroquinone increased both basal and tetanic [Ca2+]i. In ten out of thirteen fibres, the presence of any of these drugs during LIS resulted in a force reduction to about 10% of the control when fibres were returned to normal Ringer solution after the hypotonic challenge. Force production was severely depressed for at least 20 min and then recovered to control levels within 120 min. 4. Neither protease inhibitors nor a scavenger of reactive oxygen species prevented the impairment of E‐C coupling. 5. It is concluded that after a period of elevated [Ca2+]i, E‐C coupling in frog skeletal muscle becomes sensitive to the mechanical stress induced by exposure to hypotonic solution. The underlying molecular basis for this remains unclear.