Cross-bridge detachment and attachment following a step stretch imposed on active single frog muscle fibres.
1. The time course of cross-bridge detachment-attachment following a step stretch was determined in single frog muscle fibres (at 4 degrees (1 and 2.1 microns sarcomere length) by imposing, under sarcomere length control by a striation follower, test step releases of various amplitudes (2-13 nm per half-sarcomere) at successive times (4-55 ms) after a conditioning stretch of approximately 4 nm per half-sarcomere. 2. The comparison with the control tension transients, elicited by releases not preceded by the conditioning stretch, shows that, early after the conditioning stretch, the quick tension recovery following small releases is depressed and the quick tension recovery following large releases is potentiated. Both effects are expected as a consequence of the strain produced in the cross-bridges by the conditioning stretch. 3. These effects disappear and the tension transient is reprimed, indicating substitution of freshly attached cross-bridges for strained cross-bridges, with a time constant of approximately 10 ms. 4. A novel multiple-exponential equation, based on the hypothesis of complete substitution of freshly attached cross-bridges for the cross-bridges that underwent the stretch, has been used to fit the whole tension transient following step stretches of different sizes (2-6 nm per half-sarcomere). For a stretch of 4 nm, the time constant of the exponential process responsible for cross-bridge detachment (tau d, 9.3 ms) almost coincides with the time constant of repriming as measured by the double-step experiments. The time constant of the exponential process representing the cumulative effects of attachment and force generation (tau 3) is 13.6 ms. 5. For stretches of different sizes the amount of quick tension recovery attributable to the reversal of the working stroke elicited by the stretches is estimated by subtracting, from the original tension transient, the contribution to tension recovery due to detachment-attachment of cross-bridges as estimated by the multiple-exponential analysis. Following this calculation, the structural change in the myosin heads responsible for the reversal of the working stroke can be 2 nm at maximum, suggesting that the elastic component in the cross-bridges is at least twice as rigid as previously thought.
Piazzesi
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Dipartimento di Scienze Fisiologiche, Università degli Studi di Firenze, Italy.
Linari
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Reconditi
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Vanzi
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F
Lombardi
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D000818:Animals; D009119:Muscle Contraction / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000502:physiology*; D011893:Rana esculenta