For the past 60 years, muscle contraction had been thought to be governed exclusively by the contractile filaments, actin, and myosin. This thinking explained most observations for concentric and isometric, but not for eccentric muscle contractions. Just over a decade ago, we discovered that eccentric contractions were associated with a force that could not be assigned to actin and myosin, but was at least in part associated with the filamentous protein titin. Titin was found to bind calcium upon activation, thereby increasing its structural stability, and thus its stiffness and force. Furthermore, there is increasing evidence that the proximal part of titin binds to actin in an activation- and force-dependent manner, thereby shortening its free length, thus increasing its stiffness and force. Therefore, we propose that muscle contraction involves three filaments, actin, myosin and titin, and that titin regulates force by binding calcium and by shortening its spring length by binding to actin.

The sarcomere length non-uniformity theory (SLNT) is a widely accepted explanation for residual force enhancement (RFE). RFE is the increase in steady-state isometric force following active muscle stretching. The SLNT predicts that active stretching of a muscle causes sarcomere lengths (SL) to become non-uniform, with some sarcomeres stretched beyond actin–myosin filament overlap (popping), causing RFE. Despite being widely known, this theory has never been directly tested. We performed experiments on isolated rabbit muscle myofibrils (n = 12) comparing SL non-uniformities for purely isometric reference contractions (I-state) and contractions following active stretch producing RFE (FE-state). Myofibrils were activated isometrically along the descending limb of the force–length relationship (mean ± 1 standard deviation (SD) = 2.8 ± 0.3 µm sarcomere−1). Once the I-state was reached, myofibrils were shortened to an SL on the plateau of the force–length relationship (2.4 µm sarcomere−1), and then were actively stretched to the reference length (2.9 ± 0.3 µm sarcomere−1). We observed RFE in all myofibrils (39 ± 15%), and saw varying amounts of non-uniformity (1 SD = 0.9 ± 0.5 µm) that was not significantly correlated with the amount of RFE, but through pairwise comparisons was found to be significantly greater than the non-uniformity measured for the I-state (0.7 ± 0.4 µm). Three myofibrils exhibited no increase in non-uniformity. Active stretching was accompanied by sarcomere popping in four myofibrils, and seven had popped sarcomeres in the I-state. These results suggest that, while non-uniformities are present with RFE, they are also present in the I-state. Furthermore, non-uniformity is not associated with the magnitude of RFE, and myofibrils that had no increase in non-uniformity with stretch still showed normal RFE. Therefore, it appears that SL non-uniformity is a normal associate of muscle contraction, but does not contribute to RFE following active stretching of isolated skeletal muscle myofibrils.

Sarcomere length (SL) instability and SL non-uniformity have been used to explain fundamental properties of skeletal muscles, such as creep, force depression following active muscle shortening and residual force enhancement following active stretching of muscles. Regarding residual force enhancement, it has been argued that active muscle stretching causes SL instability, thereby increasing SL nonuniformity. However, we recently showed that SL non-uniformity is not increased by active muscle stretching, but it remains unclear if SL stability is affected by active stretching. Here, we used single myofibrils of rabbit psoas muscle and measured SL non-uniformity and SL instability during isometric contractions and for isometric contractions following active stretching at average SLs corresponding to the descending limb of the force-length relationship. We defined isometric contractions as contractions during which mean SL remained constant. SL instability was quantified by the rate of change of individual SLs over the course of steady-state isometric force and SL non-uniformity was defined as deviations of SLs from the mean SL at an instant of time. We found that whereas the mean SL remained constant during isometric contraction, by definition, individual SLs did not. SLs were more stable in the force-enhanced, isometric state following active stretching compared with the isometric reference state. We also found that SL instability was not correlated with the rate of change of SL non-uniformity. Also, SL non-uniformity was not different in the isometric and the post-stretch isometric contractions. We conclude that since SL is more stable but similarly non-uniform in the force-enhanced compared with the corresponding isometric reference contraction, it appears unlikely that either SL instability or SL non-uniformity contribute to the residual force enhancement property of skeletal muscle.

hyperpolarization the channel modulation was arranged as follows: Kv7>> TTX-sensitive VGSC=KATP>HCN>KNA>TTX-resistant VGSCRCav3; while by the ability to induce depolarization the sequence was Kv7=4-APsensitive Kv>>KNA>K2P>>Cav3>>KATP. The strongest and most consistent effect on Em was achieved by manipulating the activity of M-channels. Thus, retigabine (M-channel enhancer) and XE991 (M-channel inhibitor) hyperpolarized and depolarized neurons by approximately 10 mV each. Both drugs had strong reciprocal effect on evoked action potential firing. Results obtained in slices confirmed those obtained from culture. Acute topical application of retigabine, ZD7288 (HCN blocker) and pinacidil (KATP channel enhancer) to DRG in vivo significantly alleviated peripherally-induced pain however, again, retigabine was the most efficacious. Conclusions: Our study deciphers a toolkit of ion channels that sets Em of nociceptive neuron somata and identifies M-channels as one of the main controllers of nociceptor's excitability.