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Dive into the research topics where Giovanni Cecchi is active.

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Featured researches published by Giovanni Cecchi.


The Journal of Physiology | 2005

Crossbridge properties investigated by fast ramp stretching of activated frog muscle fibres

M. Angela Bagni; Giovanni Cecchi; Barbara Colombini

Very fast ramp stretches at 9.5–33 sarcomere lengths s−1 (l0s−1) stretching speed, 16–25 nm per half‐sarcomere (nm hs−1) amplitude were applied to activated intact frog muscle fibres at tetanus plateau, during the tetanus rise, during the isometric phase of relaxation and during isotonic shortening. Stretches produced an almost linear tension increase above the isometric level up to a peak, and fell to a lower value in spite of continued stretching, indicating that the fibre became suddenly very compliant. This suggests that peak tension (critical tension, Pc) represents the tension at which crossbridges are forcibly detached by the stretch. The ratio of Pc to the isometric tension at tetanus plateau (P0) was 2.37 ± 0.12 (s.e.m.). This ratio did not change significantly at lower tension (P) during the tetanus rise but decreased with time during the relaxation and increased with speed during isotonic shortening. At tetanus plateau Pc occurred when sarcomere elongation attained a critical length (Lc) of 10.98 ± 0.13 nm hs−1, independently of the stretching speed. Lc remained constant during the tetanus rise but decreased on the relaxation and increased during isotonic shortening. Length‐clamp experiments on the relaxation showed that the lower values of Pc/P ratio and Lc, were both due to the slow sarcomere stretching occurring during this phase. Our data show that Pc can be used as a measure of crossbridge number, while Lc is a measure of crossbridge mean extension. Accordingly, for a given tension, crossbridges on the isometric relaxation are fewer than during the rise, develop a greater individual force and have a greater mean extension, while during isotonic shortening crossbridges are in a greater number but develop a smaller individual force and have a smaller extension.


Biophysical Journal | 1986

Stiffness and force in activated frog skeletal muscle fibers

Giovanni Cecchi; Peter J. Griffiths; Stuart R. Taylor

Single fibers, isolated intact from frog skeletal muscles, were held firmly very near to each end by stiff metal clasps fastened to the tendons. The fibers were then placed horizontally between two steel hooks inserted in eyelets of the tendon clasps. One hook was attached to a capacitance gauge force transducer (resonance frequency up to approximately 50 kHz) and the other was attached to a moving-coil length changer. This allowed us to impose small, rapid releases (complete in less than 0.15 ms) and high frequency oscillations (up to 13 kHz) to one end of a resting or contracting fiber and measure the consequences at the other end with fast time resolution at 4 to 6 degrees C. The stiffness of short fibers (1.8-2.6 mm) was determined directly from the ratio of force to length variations produced by the length changer. The resonance frequency of short fibers was so high (approximately 40 kHz) that intrinsic oscillations were not detectably excited. The stiffness of long fibers, on the other hand, was calculated from measurement of the mechanical resonance frequency of a fiber. Using both short and long fibers, we measured the sinusoids of force at one end of a contracting fiber that were produced by relatively small sinusoidal length changes at the other end. The amplitudes of the sinusoidal length changes were small compared with the size of step changes that produce nonlinear force-extension relations. The sinusoids of force from long fibers changed amplitude and shifted phase with changes in oscillation frequency in a manner expected of a transmission line composed of mass, compliance, and viscosity, similar to that modelled by (Ford, L. E., A. F. Huxley, and R. M. Simmons, 1981, J. Physiol. (Lond.), 311:219-249). A rapid release during the plateau of tetanic tension in short fibers caused a fall in force and stiffness, a relative change in stiffness that putatively was much smaller than that of force. Our results are, for the most part, consistent with the cross-bridge model of force generation proposed by Huxley, A. F., and R. M. Simmons (1971, Nature (Lond.), 213:533-538). However, stiffness in short fibers developed markedly faster than force during the tetanus rise. Thus our findings show the presence of one or more noteworthy cross-bridge states at the onset and during the rise of active tension towards a plateau in that attachment apparently is followed by a relatively long delay before force generation occurs. A set of equations is given in the Appendix that describes the frequency dependence of the applied sinusoid and its response. This model predicts that frequency dependent changes can be used as a measure of a change in stiffness.


The Journal of Physiology | 1995

Absence of mechanical evidence for attached weakly binding cross‐bridges in frog relaxed muscle fibres.

Maria Angela Bagni; Giovanni Cecchi; F. Colomo; P. Garzella

1. Passive force responses to ramp stretches at various velocities were measured in intact and skinned single muscle fibres isolated from the lumbricalis muscle of the frog. Force was measured using a fast capacitance transducer and sarcomere length was measured using a laser light diffraction technique at a point very close to the fixed end so as to avoid effects of fibre inertia. Experiments were performed at 15 degrees C with sarcomere length between 2.13 and 3.27 microns under high (170 mM) and low (20 mM) ionic strength. 2. The analysis shows that the force response is the sum of at least three components: (i) elastic (force proportional to the amount of stretch), (ii) viscous (force proportional to rate of stretch), and (iii) viscoelastic (resembling the response of a pure viscous element in series with an elastic element). 3. The amplitude of all these components increased progressively with sarcomere length in the whole range measured. 4. A further component, attributable to the short‐range elasticity (SREC), was present in the force response of the intact fibres. 5. The amplitude of the force response decreased substantially upon skinning at high ionic strength but increased again at low ionic strength. The SREC was completely abolished by skinning. 6. None of the components of the force response was found to have the properties expected from the previously postulated ‘weakly binding bridges’.


The Journal of Physiology | 1994

Development of stiffness precedes cross-bridge attachment during the early tension rise in single frog muscle fibres.

Maria Angela Bagni; Giovanni Cecchi; F. Colomo; P. Garzella

1. Force responses to ramp stretches were recorded in single muscle fibres isolated from the lumbricalis muscle of the frog. Stretches were applied at rest and at progressively increasing times after a single stimulus. 2. The increase of fibre stiffness that precedes tension development has a ‘static’ component that accounts for the whole fibre stiffness increase during the latent period and at very low tension at the beginning of the twitch. 3. Static stiffness increase was not affected by 2,3‐butanedione‐2‐monoxime, a drug that almost completely inhibited twitch tension. 4. Static stiffness increased approximately 5‐fold as the sarcomere length was increased from 2.1 to 2.84 microns. 5. These results suggest that static fibre stiffness increase is not attributable to the formation of non‐force‐generating cross‐bridges.


Journal of Muscle Research and Cell Motility | 1990

Tension and stiffness of frog muscle fibres at full filament overlap.

Maria Angela Bagni; Giovanni Cecchi; F. Colomo; Corrado Poggesi

SummaryStiffness measurements in activated skeletal muscle fibres are often used as one means of estimating the number of attached crossbridges on the assumption that myofilament compliances do not contribute significantly to the fibre compliance. This assumption was tested by studying the effects of sarcomere length on fibre stiffness in the plateau region of the length-tension diagram (from 1.96 to 2.16μm sarcomere length in the tibialis anterior muscle of the frog). Lengthening of the sarcomere across this region in fact, produces only an increase in the proportion of actin filament free from cross-bridges without altering the amount of effective overlap; no change in fibre stiffness is therefore expected if actin filaments are perfectly rigid. The results show that while tetanic tension remained constant within 1.5%, as the sarcomere length was increased from 1.96 to 2.16μm fibre stiffness decreased by about 4%, indicating that a significant proportion of sarcomere compliance is localized in the actin filaments. A simple model based on the sliding filament theory was used in order to calculate the relative contribution of actin filaments to fibre compliance. In the model it was assumed that fibre compliance resulted from the combination of crossbridge compliance (distributed over the overlap zone) in series with thin filament and tendon compliances. The calculations show that the experimental data could be adequately predicted only assuming that about 19% of sarcomere compliance is due to actin filament compliance.


Journal of Muscle Research and Cell Motility | 1992

Effects of 2,3-butanedione monoxime on the crossbridge kinetics in frog single muscle fibres.

Maria Angela Bagni; Giovanni Cecchi; F. Colomo; P. Garzella

SummaryThe effects of 2,3-butanedione monoxime (BDM) on contraction characteristics were studied at 5‡C in single intact fibres isolated from the tibialis anterior muscle of the frog. The force-velocity relation was determined using the controlled-velocity method in either whole fibres or short fibre segments in which sarcomere shortening was measured by a laser light diffraction method. It is shown that 3mm BDM decreases the speed of rise and the amount of tetanus tension, reduces the maximum velocity of shortening and increases the curvature of the force-velocity relation, as well as the value for the stiffness to tension ratio. BDM also slowed down the redevelopment of tetanus tension after a period of unloaded shortening both in fixed-end and in length-clamp conditions. In normal and in BDM-treated fibres length-clamping increased the speed of the initial rise of tetanus tension but not that of the recovery after shortening. The observed force-velocity data points were fitted by the Huxley (1957) equation. It was found that BDM produces a conspicuous decrease of the rate constant for crossbridge attachment. This effect, and also a reduction of the force per crossbridge, are responsible for the depression of the contractile characteristics produced by BDM.


The Journal of Physiology | 1988

Plateau and descending limb of the sarcomere length-tension relation in short length-clamped segments of frog muscle fibres

Maria Angela Bagni; Giovanni Cecchi; F. Colomo; Chiara Tesi

1. The relation between sarcomere length and tetanic tension was determined at 10‐12 degrees C for 70‐80 microns long segments of single fibres isolated from the tibialis anterior and semitendinosus muscles of the frog. Measurements of segment striation spacings were performed during fixed‐end or length‐clamp contractions by means of a laser light diffractometer. 2. At sarcomere lengths of around 2.10 microns tetanic tension rose promptly to a steady plateau, independent of the recording conditions. At greater sarcomere lengths under fixed‐end conditions the tension rise occurred in two distinct stages: an initial rapid rise followed by a much slower creep. The tension creep was entirely abolished in length‐clamp contractions. 3. The sarcomere length‐tension diagram of length‐clamped segments of tibialis anterior fibres exhibited a definite flat region between about 1.96 and 2.16 microns where tension varied by less than 1.5%. The highly linear descending limb reached zero tension at about 3.53 microns. The shift to the left by about 0.10 microns, with respect to the length‐tension diagram of length‐clamped segments of semitendinosus fibres, may be tentatively explained by assuming that thin filament lengths vary in different muscles. 4. The results are in agreement with those of a previous work by Gordon, Huxley & Julian (1966) and support the hypothesis (Huxley, 1957, 1980) that muscle tension is produced by simultaneous action of independent force generators, in proportion to the number of myosin bridges overlapped by actin filaments.


The Journal of Physiology | 1990

Myofilament spacing and force generation in intact frog muscle fibres.

Maria Angela Bagni; Giovanni Cecchi; F. Colomo

1. The relation between sarcomere length and steady tetanic tension was determined at 10‐12 degrees C for 70‐80 microns long length‐clamped segments of single fibres isolated from the tibialis anterior muscle of the frog, in normal and hypertonic or hypotonic Ringer solutions. 2. The tension depression and potentiation observed in hypertonic and hypotonic Ringers solutions varied with sarcomere length, so that, as opposed to myofilament overlap predictions, the optimum length for tension development was shorter in hypertonic Ringer solution and longer in hypotonic Ringer solution than in normal Ringer solution. As the fibres were stretched from 1.96 to 2.24 microns sarcomere length, both tension depression in hypertonic Ringer solution and tension potentiation in hypotonic Ringer solution increased by 9 and 5%, respectively. 3. Within this range of sarcomere lengths the length‐stiffness relation in hypotonic and in hypertonic Ringer solutions exhibit little or no change relative to that in normal Ringer solution. 4. The results indicate that separation between the thick and the thin myofilaments influences the mechanism of force generation. There is an optimum interfilament distance (10‐12 nm surface to surface between the thick and the thin filaments) for tension production. In isotonic Ringer solution, this corresponds to the interfilament distance at sarcomere lengths around 2.10 microns. The force per attached cross‐bridge, rather than their number, appears to decrease as the interfilament distance is brought above or below the optimum length. Even if this effect is moderate in isotonic Ringer solution, it should be taken into account in models of the force‐generation mechanism.


Journal of Muscle Research and Cell Motility | 2004

Force responses to fast ramp stretches in stimulated frog skeletal muscle fibres

Maria Angela Bagni; Giovanni Cecchi; E. Cecchini; Barbara Colombini; F. Colomo

Force responses to fast ramp stretches at various velocities were recorded from single muscle fibres isolated from either lumbricalis digiti IV or tibialis anterior muscle of the frog (Rana esculenta) at sarcomere length between 2.15 and 3.25 μm at 15° C. Stretches were applied at rest, at tetanus plateau and during the tetanus rise. Stretches with the same velocity but different accelerations were imposed to the fibre to evaluate the effect of fibre inertia on the force responses. Length changes were measured at sarcomere level with either a laser diffractometer or a striation follower apparatus. The force response to a fast ramp stretch could be divided into two phases. The initial fast one (phase 1) lasts for the acceleration period during which the stretching velocity rises up to the steady state. The second slower phase (phase 2) lasts for the remainder of the stretch and corresponds to the well-known elastic response of the fibre. Most of this paper is concerned with phase 1. The amplitude of the initial fast phase was proportional to the stretching velocity as expected from a viscous response. This viscosity was associated with a very short (about 10 μs) relaxation time. The amplitude of the fast phase increased progressively with tension during the tetanus rise and scaled down with sarcomere length approximately in the same way as tetanic tension and fibre stiffness. These data suggest that activated fibres have a significant internal viscosity which may arise from crossbridge interaction


The Journal of Physiology | 2007

Crossbridge properties during force enhancement by slow stretching in single intact frog muscle fibres.

Barbara Colombini; Marta Nocella; Giulia Benelli; Giovanni Cecchi; Maria Angela Bagni

The mechanism of force enhancement during lengthening was investigated on single frog muscle fibres by using fast stretches to measure the rupture tension of the crossbridge ensemble. Fast stretches were applied to one end of the activated fibre and force responses were measured at the other. Sarcomere length was measured by a striation follower device. Fast stretching induced a linear increase of tension that reached a peak and fell before the end of the stretch indicating that a sudden increase of fibre compliance occurred due to forced crossbridge detachment induced by the fast loading. The peak tension (critical tension, Pc) and the sarcomere length needed to reach Pc (critical length, Lc) were measured at various tensions during the isometric tetanus rise and during force enhancement by slow lengthening. The data showed that Pc was proportional to the tension generated by the fibre under both isometric and slow lengthening conditions. However, for a given tension increase, Pc was 6.5 times greater during isometric than during lengthening conditions. Isometric critical length was 13.04 ± 0.17 nm per half‐sarcomere (nm hs−1) independently of tension. During slow lengthening critical length fell as the force enhancement increased. For 90% enhancement, Lc reduced to 8.19 ± 0.039 nm hs−1. Assuming that the rupture force of the individual crossbridge is constant, these data indicate that the increase of crossbridge number during lengthening accounts for only 15.4% of the total force enhancement. The remaining 84.6% is accounted for by the increased mean strain of the crossbridges.

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F. Colomo

University of Florence

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P. Garzella

University of Florence

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Chiara Tesi

University of Florence

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