电磁场对青蛙骨骼肌单根肌纤维效应的研究

电磁场对完整和去膜青蛙肌纤维作用的比较研究表明,交变电场通过改变膜电位引起肌肉收缩,在此过程中收缩蛋白质的空间位置而非自身构象发生变化,横桥尤其是S－2片段,在伴随横桥从弱耦合状态向强耦合状态过渡时远离粗肌丝而向细肌丝运动,使其与粗肌丝骨架的平均取向比松弛状态或静息状态时相对增大．一般强度恒定磁场对肌纤维膜电位状态及肌纤维内部蛋白质分子的运动及其相互作用影响极其微弱．     

鱼类肌肉组织发生和分化相关基因的研究进展

鱼类骨骼肌由两种分布位置不同的肌纤维组成,一种是位于皮下浅层的慢收缩红肌纤维,另一种是构成躯体绝大部分的快收缩白肌纤维.肌肉组织中含有多种不同类型的蛋白质如具有收缩功能的结构蛋白,可溶性的肌肉蛋白及肌肉特异性的转录因子等,这些所有的肌源蛋白的产生都经过了复杂的肌肉发生与分化过程.基于相关研究近况,分析了多种转录因子的正、负向调控对肌肉分化发育的影响,同时讨论了研究此调控过程的理论价值及未来应用意义.     

单根青蛙肌纤维椭偏特性的温度效应的初步研究

利用衍射光椭圆偏振参数的变化研究温度(6℃至40℃)对单根青蛙肌纤维内部结构的影响.实验测得,当温度从6℃上升到20℃左右时,衍射光的退偏率r缓慢下降,20℃后r随温度升高而增大,即在20℃左右处出现一双向变化特性;当温度超过30℃后r急剧减小,在30℃附近呈现一极大值.若温度高于35℃左右时,被肌纤维散射的衍射光消失.由此表明温度变化不仅会导致横桥空间取向改变,而且会引起横桥动力学性质的变化;温度过高时,蛋白质分子的热运动会造成肌纤维内部有序结构的破坏.     

三叉神经对去运动神经支配面肌肌萎缩的影响

本文用组织化学、电镜以及肌球蛋白和肌动蛋白电泳分析了在单纯性面神经切断和三叉神经、面神经同时切断后面肌萎缩的病理改变--肌纤维的显微结构改变以及面肌收缩蛋白质在手术后不同时期的降解变化。实验证明,面部肌肉在不同的神经切断情况下其病理改变不同,正常的三叉神经支配可以延缓面瘫后肌肉蛋白质的降解,减少胶原纤维结缔组织的增生,较好地保护肌纤维的显微结构,延缓和减轻去运动神经支配面肌肌萎缩。本研究结果可以为临床治疗面瘫提供一定的理论指导。     

去负荷小鼠比目鱼肌收缩特性和纤维类型转化

目的:探讨去负荷后小鼠比目鱼肌的收缩特性与骨骼肌纤维类型转化之间的关系。方法:采用离体肌肉灌流技术和电刺激方法,在小鼠后肢去负荷28 d引起骨骼肌萎缩后,观察比目鱼肌单收缩、强直收缩能力和肌疲劳指标等收缩特性的改变,同时利用组织免疫荧光染色和实时定量聚合酶链式反应(real-time PCR)等技术检测去负荷后比目鱼肌快慢肌纤维组成和纤维类型转化的变化。结果:去负荷28 d后,小鼠比目鱼肌单收缩力、强直收缩能力和疲劳指数(fatigue index)均有显著性下降,同时伴有快肌纤维亚型的增加和慢肌纤维亚型的减少。结论:去负荷28 d后小鼠比目鱼肌收缩特性的改变和快慢肌纤维类型的转化有关。     

单根青蛙肌肉纤维钙离子活化收缩过程的椭园偏振测量的研究

本文介绍了用椭圆偏振法对单根青蛙肌肉纤维钙离子活化收缩机制的研究.实验发现,衍射光所提供的椭圆偏振谱是肌小节长度的函数,去膜肌纤维的总双折射系数△n(?)随着肌小节长度的增大而增大,而退偏系数rт则在肌小节长度2.9μm附近处出现一个极大值.当肌纤维受钙离子作用活化收缩时,在肌小节长度2.9μm附近,rт的相对变化量最大,即(△r/r0)=5l.3%.上述结果表明肌动蛋白丝和肌球蛋白丝之间的重叠区和非重叠区中横桥的平均取向是不同的,在钙离子作用活化收缩时,会导致肌动蛋白丝和肌球蛋白丝之间的质量重新分配.     

大鼠快、慢肌纤维肌球蛋白的特征

根据生化和收缩功能特性的研究,肌纤维可分为三种基本类型:慢收缩氧化型,快收缩氧化、醣酵解型和快收缩醣酵解型。由于收缩快慢不同,要求对ATP有不同的水解速率,这样,快慢二类纤维的肌球蛋白可能也会存在差别。当然,从对肌球蛋白轻链的分析我们已经知道,在快、慢肌之间轻链是有区别的。但是轻链的特征还未用来区别快、慢肌纤维类型;而用测酶系活力的方法来研究纤维类型,因各种酶活力又互相重迭,使分析结果复杂化,从鉴别的角度来考虑也不够满意。如果我们用单肌纤维为材料分析其完整肌     

蟾蜍横纹肌在硫脲作用下兴奋和收缩的分离

蟾蜍缝匠肌在含0.7M 硫脲的任氏溶液中浸泡后,丧失对直接刺激作电反应和机械反应的能力。将肌肉放回正常任氏溶液后,电反应能力能逐渐恢复,而机械反应能力则不能,由此可以得到表现兴奋与收缩分离的肌肉标本。硫脲不仅能使活的肌肉失去收缩能力,还能使经甘油抽提过的肌纤维束不可逆地丧失因三磷酸腺苷作用而缩短的能力。经甘油抽提的肌纤维束因三磷酸腺苷的作用而缩短后,可因再用硫脲溶液处理而伸长。以上这些结果指示硫脲对肌肉收缩能力的影响是对收缩蛋白体系直接作用的结果。肌纤维经硫脲处理后膜电位下降,动作电位振幅减低,时程变慢,膜电阻和膜电容都减小。硫脲对神经肌肉接头传递也能产生可逆的阻滞。     

肌纤维类型与运动能力

目前,关于肌纤维的分类和命名尚有争论。劳伦悉尼(Lorazini,1673)首次报道兔骨胳肌有红、白之分,并发现肌肉的色泽与运动能力有关。兰威尔(Ranvier,1883)用电刺激法证明红肌收缩慢而持久,白肌收缩快却不能持久,从而提出将骨胳肌纤维分为“红肌”和“白肌”两种类型。近年来,由于针刺活检肌组织法的采用和超微量分析技术的发展,对肌纤维的理化特性、生理功能等进行了广泛的研究,发现红肌不一定都是收缩慢的,故将肌纤维分为“慢肌”     

电场中的蛋白质结晶

人们一直致力于寻求提高蛋白质晶体质量的方法,利用电场诱导蛋白质结晶即是诸多方法中的一种。已有文献报道显示,电场对蛋白质结晶的影响是积极的。我们从直流电场、交流电场、内置电场、外置电场对蛋白质结晶的影响及相关结晶设备,电场中生长的蛋白质晶体质量的评估,电场中蛋白质结晶的原理及影响因素等方面,对已报道的电场中的蛋白质结晶研究工作进行了总结。     

Calcium-induced and voltage-dependent inactivation of calcium channels in crab muscle fibres

Inactivation of Ca channels was examined in crab muscle fibres using the voltage-clamp method. A satisfactory suppression of outward currents was attempted by the use of K+ blocking agents: TEA, 4AP and Cs ions instead of K+ ions applied extracellularly. The inactivation of Ca current appeared as a bi-exponential process. The faster component had a mean value of the time constant of 50 ms while the second component inactivated at a tenfold slower rate. The extent of inactivation of the faster component increased as the Ca current itself increased in different experimental conditions. Inactivation decreased when ICa was reduced for large applied depolarizations. The time constant of the faster calcium component also depended on the calcium current. Thus the results suggested that Ca2+ entry leads to inactivation of one component of calcium current in crab muscle. Substitution of Ca2+ ions by Sr2+ or Ba2+ ruled out the hypothesis concerning an accumulation process which would explain the decrease of the inward current. The second slower component of Ca current was better described by a voltage-dependent mechanism and its rate was not modified in Ca2+ rich solution or when the inward current was carried by Sr2+ or Ba2+ ions. Thus in crab muscle fibres, inactivation is mediated by both calcium entry and a voltage-gated mechanism.     

细胞离子在振荡电磁场作用下的受力模型分析

本文通过生物细胞模型,研究振荡电场、振荡磁场以及振荡磁场产生的感应电场对细胞离子的作用机理。模型分析结果表明,电场力和罗仑兹力对细胞膜两侧的自由离子将产生加速度,振荡离子将产生周期性电位移。该模型同时也解释了脉冲电磁场比同参教的连续场产生更多的生物效应,以及连续场在开始施加和切除时的效应最大。     

Characteristics of functioning of electromechanical coupling in striated muscles of higher and lower vertebrates

A comparative pharmacological analysis of relative contributions of different signal transduction pathways in the activation of contraction (excitation-contraction coupling, ECC) in intact fast striated muscles of frog and lamprey was performed. It was found that the major mechanism responsible for the ECC in muscles of both animals is Ca2+ release from the sarcoplasmic reticulum through the ryanodine-sensitive channels. However, the ECC in lamprey muscle displays some important differences in the units of electromechanical coupling, which precede the calcium release from sarcoplasmic reticulum. The maximum contraction force in frog muscle develops during caffeine-induced contracture, which indicates that all Ca2+ stored in sarcoplasmic reticulum is released through ryanodine-sensitive channels. In contrast, in lamprey muscle, the maximum force develops not in response to high caffeine concentration, but in response to repetitive electrical stimulation. Hence, in addition to stores liberated by ryanodine-sensitive channels, some other sources of calcium ions should exist, which contribute to the contraction activation. A source of this additional Ca2+ ions can be external medium, because acetylcholine contracture is abolished in a calcium-free medium. In frog muscle, the acetylcholine contracture was abolished in a Na(+)-free solution. It was concluded that in frog muscle ECC can be triggered by changes in the transmembrane potential (depolarization-induced calcium release), while in lamprey muscle the entry of calcium ions into myoplasm as the trigger in ECC (calcium-induced calcium release). The lamprey muscle was found to be more resistant to tetrodotoxin and tetracaine, which is indicative of a role in the activation of contraction of tetrodotoxin-resistant Na+ and/or Ca2+ channels. It was concluded, that ECC mechanism in striated muscles of low vertebrates is not limited by the generally accepted scheme of depolarization-induced calcium release but can include some other schemes, which require the Ca2+ influx into the cell.     

Measurement of sarcoplasmic reticulum Ca2+ content in intact amphibian skeletal muscle fibres with 4-chloro-m-cresol.

Single skeletal muscle fibres were isolated from the toad (Bufo marinus) and isometric force and myoplasmic free calcium concentration ([Ca2+]i) were measured. Brief applications of 4-chloro- m-cresol (4-CmC, 0.2-5 mM) elevated [Ca2+]i reversibly in a dose-dependent manner. The lowest concentration of 4-CmC which reliably gave maximal [Ca2+]i was 2 mM and it was, therefore, used for measurement of sarcoplasmic reticulum (SR) Ca2+ content. Tetanic stimulations (100 Hz) increased [Ca2+]i from a resting level of 105 +/- 47 nM (n = 10) to 1370 +/- 220 nM (n = 6). Application of 2 mM 4-CmC produced a contracture that was 54 +/- 16% (n = 6) of the tetanic force and elevated [Ca2+]i to a peak of 3520 +/- 540 nM (n = 8). Both force and [Ca2+]i levels (resting and tetanic) were restored after 10 min of washout of 4-CmC. In skinned muscle fibres, the myofibrillar Ca(2+)-sensitivity was not changed by 4-CmC, but maximal force was reduced to 74 +/- 10% (n = 4). The magnitude of the peak of the 4-CmC-induced Ca2+ transient was not significantly changed by removal of extracellular Ca2+ nor by inhibiting the SR Ca2+ pump with 2,5-di-tert-butylhydroquinone. Treatment of intact fibres with 30 mM caffeine produced a peak Ca2+ level that was indistinguishable from 2 mM 4-CmC. These results indicate that it is possible to measure the SR Ca2+ content in the same fibre with 4-CmC without loss of normal muscle function.     

Physics in muscle research

Muscle is one of few organs whose performance can be measured by physical quantities. However, very few attempts have been made to apply theoretical physics to muscle. In this paper we will see how physical principles can be applied by taking advantage of unique properties of muscle structure. The first topic is to establish the stability conditions of sarcomere structure. The conclusions are then compared to some experimental facts. Next, we move on to the field theory fundamentals. The concept of energy density as a stress tensor is shown to be a powerful tool for the dielectric force theory to understand how proteins move under electric fields. By combining the structural stability theory and the dielectric force theory we arrive at a helical dipole array. We discuss the source of strong dipole fields and how the dipole strength could be controlled by Ca ions. The behavior of water and ions under electric fields is briefly discussed. The third topic is the mechanical stiffness of muscle in longitudinal and lateral directions. Some experimental data are shown and the physics of anisotropic stiffness is discussed. An appendix is provided to explain the pitfalls of experimenting with isolated components rather than organized structures (sarcomere).     

Calcium and electropermeabilized cells

Trains of short and intense electric pulses may induce a reversible local permeabilization on the membrane of the treated cells. Hydrophilic species can then almost freely cross the envelope and either enter or escape from the cytoplasm. The purpose of the present study was to investigate the possibility of introducing well defined amounts of Ca2+ ions within the cell. Chinese hamster ovary cells were used as a model system. When the pulsing buffer contained high levels of free Ca2+, the survival of cells was strongly affected. A 1 mM level was well tolerated. When cells were pulsed under moderated field conditions, it was observed that Ca2+ entered cells very rapidly (second time range). But the basic cytoplasmic level was set back spontaneously within a few minutes. The perspectives of this electrical injection are discussed for basic cell biology and high-throughput biotechnology.     

Morphometric properties and innervation of muscle compartments in rat medial gastrocnemius

The rat medial gastrocnemius (MG) muscle is composed of the proximal and distal compartments. In this study, morphometric properties of the compartments and their muscle fibres at five levels of the muscle length and the innervation pattern of these compartments from lumbar segments were investigated. The size and number of muscle fibres in the compartments were different. The proximal compartment at the largest cross section (25% of the muscle length) had 34% smaller cross-sectional area but contained a slightly higher number of muscle fibres (max. 5521 vs. 5360) in comparison to data for the distal compartment which had the largest cross-sectional area at 40% of the muscle length. The muscle fibre diameters revealed a clear tendency within both compartments to increase along the muscle (from the knee to the Achilles tendon) up to 46.9?μm in the proximal compartment and 58.4?μm in the distal one. The maximal tetanic and single twitch force evoked by stimulation of L4, L5, and L6 ventral roots in whole muscle and compartments were measured. The MG was innervated from L4 and L5, only L5, or L5 and L6 segments. The proximal compartment was innervated by axons from L5 or L5 and L4, and the distal one from L5, L5 and L6, or L5 and L4 segments. The forces produced by the compartments summed non-linearly. The tetanic forces of the proximal and distal compartments amounted to 2.24 and 4.86?N, respectively, and their algebraic sums were 11% higher than the whole muscle force (6.37?N).     

The kinetics relating calcium and force in skeletal muscle.

The kinetics relating Ca2+ transients and muscle force were examined using data obtained with the photoprotein aequorin in skeletal muscles of the rat, barnacle, and frog. These data were fitted by various models using nonlinear methods for minimizing the least mean square errors. Models in which Ca2+ binding to troponin was rate limiting for force production did not produce good agreement with the observed data, except for a small twitch of the barnacle muscle. Models in which cross-bridge kinetics were rate limiting also did not produce good agreement with the observed data, unless the detachment rate constant was allowed to increase sharply on the falling phase of tension production. Increasing the number of cross-bridge states did not dramatically improve the agreement between predicted and observed force. We conclude that the dynamic relationship between Ca2+ transients and force production in intact muscle fibers under physiological conditions can be approximated by a model in which (a) two Ca2+ ions bind rapidly to each troponin molecule, (b) force production is limited by the rate of formation of tightly bound cross-bridges, and (c) the rate of cross-bridge detachment increases rapidly once tension begins to decline and free Ca2+ levels have fallen to low values after the last stimulus. Such a model can account not only for the pattern of force production during a twitch and tetanus, but also the complex, nonlinear pattern of summation which is observed during an unfused tetanus at intermediate rates of stimulation.     

Dynamic changes of [Ca~(2 )]_i in cerebellar granule cells exposed to pulsed electric fields

Intracellular free Ca2 concentration ([Ca2 ]i) in embryonic chick cerebellar granule cells loaded with fluo-3/AM and exposed to a single pulsed electric field was investigated using a confocal laser scanning microscope and fluorescent microscope equipped with CCD video imaging system. The results showed that [Ca2 ]i increased immediately and rose to the peak rapidly as the cells exposed to a single pulsed electric field. The amplitude and rate of the increases of [Ca2 ]i depend on the intensity of external electric field. In the presence of Ca2 chelant EGTA or Ca2 channels blocker La3 in the pulsation solutions, the increase of [Ca2 ]i was still observable. It was also observed that [Ca2 ]i of different intracellular areas in the cell elevated simultaneously while the peak of the increase of [Ca2 ]i in the poles of the cell preceded to the peak in its somata and recovered to a plateau within a short time.