Muscle stretching induces twitch contractions without activation of stretch-activated channels in intact rat trabeculae

Introduction Mechano-electric coupling (MEC) means that muscle stretching can induce action potentials. Stretch-activated channels (SACs) have been believed to play important roles in their induction. Purpose To investigate what degree of muscle stretching can induce MEC-mediated action potentials and what roles SACs play in their induction. Methods Trabeculae were obtained from right ventricles of rat hearts. Force was measured with a strain gauge, sarcomere length (SL) with a laser diffraction technique, and [Ca2+]i with fura-2 (24°C). The SL was set at 2.0 μm at the resting condition. Trabeculae were stimulated electrically at 400-ms intervals for 7.5 s. Various degrees of muscle stretching were applied at 500 ms after the last stimulus of the electrical train to determine the minimal SL (SL-AP) at which an action potential or a twitch contraction was induced by the stretching (0.7 mM [Ca2+]o). Results The SL-AP was 2.34±0.02 μm (n=8) when trabeculae were stretched rapidly from a SL of 2.0 μm (400-ms stimulation intervals, 0.7 mM [Ca2+]o). The SL-AP was not changed by increasing the stimulation intervals from 400 to 2000 ms (n=7), by increasing [Ca2+]o from 0.7 to 2 mM (n=8), and by adding 1 μM isoproterenol (n=8), suggesting that Ca2+ loading within the myocardium has no effect on the SL-AP. Surprisingly, the SL-AP was not changed by adding 5 μM GsMTx4 (n=8), 10 mM Gd3+ (n=9), 100 μM (n=8) and 200 μM streptomycin (n=11), revealing that SACs play no roles in the determination of SL-AP. The SL-AP was not changed by adding 1 μM ryanodine (n=5) and 30 μM cyclopiazonic acid and was not changed by adding 3 μM diphenyleneiodonium chloride (n=8) and 10 μM colchicine, suggesting that Ca2+ leak from the SR and activation of NADPH oxidase has no effect on the SL-AP. In contrast, elevation of temperature from 23 to 36°C decreased the SL-AP from 2.35±0.01 to 2.34±0.02 μm (p<0.05, n=7). Elevation of extracellular K+ ([K+]o) from 5 to 10 mM increased the SL-AP from 2.35±0.01 to 2.38±0.01 μm (p<0.01, n=7), while reduction of [K+]o to 5 mM decreased it to 2.36±0.01 μm (p<0.05, n=7), suggesting that depolarization of membrane potential suppresses MEC-mediated twitch contractions. The SL-AP was increased from 2.34±0.01 to 2.36±0.01 μm (p<0.01, n=7) when stretching was applied at a shorter interval after the last stimulus, i.e., 200 ms. After electrical stimulation at 300-ms stimulation intervals for 30 s, arrhythmias were induced by a MEC-mediated twitch contraction in 6 out of 9 trabeculae when stretching was applied at 500 ms after the last stimulus, while they were induced only in 2 out of 9 trabeculae without the stretching (4 mM [Ca2+]o, 1 μM isoproterenol). Conclusions These results suggest that muscle stretching causes membrane excitation, which potentially induces arrhythmias and that activation of SACs, Ca2+ release from the SR, and activation of NADPH oxidase by muscle stretching are not involved in the excitation. FUNDunding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Grant-in-Aid for Scientific Research (C) from Japan Society for the Promotion of Science.

Neural and muscular determinants of maximal rate of force development

The ability to produce rapid forces requires quick motor unit recruitment, high motor unit discharge rates, and fast motor unit force twitches. The relative importance of these parameters for maximum rate of force development (RFD), however, is poorly understood. In this study, we systematically investigated these relationships using a computational model of motor unit pool activity and force. Across simulations, neural and muscular properties were systematically varied in experimentally observed ranges. Motor units were recruited over an interval starting from contraction onset (range: 22–233 ms). Upon recruitment, discharge rates declined from an initial rate (range: 89–212 pulses per second), with varying likelihood of doublet (interspike interval of 3 ms; range: 0–50%). Finally, muscular adaptations were modeled by changing average twitch contraction time (range: 42–78 ms). Spectral analysis showed that the effective neural drive to the simulated muscle had smaller bandwidths than the average motor unit twitch, indicating that the bandwidth of the motor output, and thus the capacity for explosive force, was limited mainly by neural properties. The simulated RFD increased by 1,050 ± 281% maximal voluntary contraction force per second from the longest to the shortest recruitment interval. This effect was more than fourfold higher than the effect of increasing the initial discharge rate, more than fivefold higher than the effect of increasing the chance of doublets, and more than sixfold higher than the effect of decreasing twitch contraction times. The simulated results suggest that the physiological variation of the rate by which motor units are recruited during ballistic contractions is the main determinant for the variability in RFD across individuals. NEW & NOTEWORTHY An important limitation of human performance is the ability to generate explosive movements by means of rapid development of muscle force. The physiological determinants of this ability, however, are poorly understood. In this study, we show using extensive simulations that the rate by which motor units are recruited is the main limiting factor for maximum rate of force development.

Abstract 20385: Pregnancy-Induced Cardiac Hypertrophy is Accompanied by Unique Contractile Properties Not Seen in Other Forms of Physiological Hypertrophy

Background: Cardiac hypertrophy accompanying pregnancy has generally been categorized as physiologic hypertrophy similar to that seen with exercise, however a reduction in cardiac function in late pregnancy has been suggested. Furthermore, the hemodynamic stress of pregnancy can induce a maladaptive, pathologic hypertrophy in a small number of women. This study seeks to characterize the contractile properties of late-pregnant myocardium. Methods and Results: Late Pregnancy (LP) Female Swiss-Webster mice were bred then studied at near term (Embryonic day 17-19) and compared to age-matched, non-pregnant (NP) controls. Individual cardiac myocytes were isolated using collagenase-based perfusion technique. Two-dimensional Surface Area measured in quiescent cells was elevated (p<.01) in LP myocytes (LPM) (3609± 132u 2 ) vs NP myocytes (NPM) (2736± 88u 2 ), and this increase was due to increases in both length (8.5%) and width (15.6%). Western Blot analysis showed a reduction in Ryanodine Receptor protein in LP, but no differences in L-type Ca Channel, SERCA or Phospholamban levels. Sarcomere length (light diffraction) and Ca 2+ transients (fluo-3) were measured at pacing rates of 1 Hz and at bath [Ca] of 2mM. Duration of twitch contraction was prolonged (p<.05) in LPM as measured by Time to 75% Recovery (.42 ± .02 vs .37 ±.01 sec in NPM) and Time to 90% recovery (.51 ± .02 vs .45 ± .02 sec in NPM). There were no differences in other contractile parameters measured or in the fluo-3 calcium transient properties. 10 -7 M Isoproterenol (ISO) was used to determine the responsiveness to adrenergic stimulation. ISO induced significantly enhanced contractility in both LPM and NPM, and the response was heightened in LPM such that the presence of ISO normalized the differences in the duration of twitch contraction between LPM and NPM. Conclusions: These results suggest that hypertrophied LPM have characteristics of both physiologic and pathologic hypertrophy including enhanced responsiveness to ISO and a prolonged relaxation phase. The prolongation of relaxation is not seen in physiologic hypertrophy induced by exercise and may contribute to the diastolic dysfunction reported in some pregnancies. Enhanced response to ISO suggests an increased cardiac reserve in LPM.

Adaptations in physiological properties of rat motor units following 5 weeks of whole-body vibration

The purpose of the study was to determine the effects of 5-week whole-body vibration (WBV) on contractile parameters and force–frequency relationship of functionally isolated motor units of the rat medial gastrocnemius muscle: fast fatigable (FF), fast fatigue-resistant (FR), and slow (S). Moreover, myosin heavy chain isoform content was quantified. Following WBV, the maximum tetanic force of FF units was increased by ∼25%. The twitch half-relaxation time in all types of motor units and the twitch contraction time in FR units were shortened. The twitch-to-tetanus force ratio was decreased and the force–frequency curves were shifted rightwards in S and FR units. Myosin heavy chain distribution was not changed. These findings suggest modifications of the excitation–contraction coupling towards shortening of a twitch contraction. The observed increase in force of FF units may contribute to gains in muscle dynamic strength reported following WBV treatment.