螯虾腹屈肌的超微结构

用电子显微镜观察,发现螯虾(Procambarus clarkii)腹屈肌浅层(慢肌,tonic fiber)肌纤维和深层(快肌,twitch fiber)肌纤维的超微结构存在显著差异。浅层腹屈肌肌原纤维有相对长的肌节(5—10μm),肌原纤维直径较大,每根粗肌丝周围有9—12根细肌丝环绕,细肌丝与粗肌丝数量比约为6∶1;深层腹屈肌有相对短的肌节(3—4.5μm),肌原纤维直径较小,每根粗肌丝周围有6根细肌丝环绕,细肌丝与粗肌丝数量比为3∶1。以上结果提示,在决定螯虾腹屈肌收缩速度方面,可能肌原纤维直径大小比肌节长度更为重要。细、粗肌丝排列方式也可能与收缩速度有关。这与脊椎动物骨骼肌的情况是不同的。     

穴居狼蛛腿肌的显微和亚显微结构

用光学显微镜、电子显微镜和小角 X 射线衍射技术研究了穴居狼蛛(Lycosa singriensis)腿肌的构造。腿肌的肌原纤维显示 I 带和 A 带构成的典型的横纹,肌节长度比脊椎动物骨胳肌长得多,约为5.3μm。A 带中间存在 H 带和 M 线区。在肌原纤维中,长4.3μm,直径200(?)的粗肌丝排成中心间距为约600(?)的六角阵列,每根粗肌丝周围环绕着8—12根直径60(?)的纽肌丝。粗肌丝中心的显著浅染提示粗肌丝可能由肌球蛋白围绕着副肌球蛋白核心组成。X射线衍射实验结果指示,静息状态活肌肉中,肌球蛋白横桥形成435(?)重复螺旋,横桥水平的轴向间距为145(?)。静息状态在位固定的腿肌的电镜照片还显示与肌动蛋白细肌丝接触的肌球蛋白横桥,文中讨论了这一颇有兴趣的结果。     

可口革囊星虫肾管肌组织的结构特征

采用显微及亚显微技术观察了可1：7革囊星虫肾管肌组织的结构特征。肾管肌组织位于柱状上皮层下,由纵肌及环肌组成。肌细胞（肌纤维）呈长梭形,核位于细胞边缘并明显突向细胞外基质中,核周围有较多线粒体及少量内质网。肌纤维表面有许多囊状或指状突起的肌质囊,内含肌浆、光面内质网、线粒体及糖原颗粒。肌质囊之间的肌膜内面具膜相关电子致密斑。肌纤维内含粗、细两种肌丝,细肌丝围绕在粗肌丝周围,在肌丝之间分布有糖原颗粒、线粒体及胞质致密体。线粒体及糖原为肌纤维的代谢提供能量,肌组织的收缩对促进肾管的过滤排泄及繁殖时配子进入肾管可能起重要作用。     

蜜蜂飞翔肌收缩蛋白的电子显微镜鉴定

自五十年代肌丝滑行模型建立以来,关于脊椎动物骨胳肌的蛋白质成分,肌丝排列以及肌肉收缩时结构变化的研究取得了很大的进展。骨胳肌肌原纤维由粗、细肌丝有规律地排列所组成。对于肌肉收缩蛋白的选择性抽提,专一性抗体标记以及重组肌丝的研究,证实肌球蛋白存在于粗肌丝;肌动蛋白、原肌球蛋白和原宁蛋白存在于细肌丝(Huxley,A.F.,1957;Huxley,H.E;,1972)。昆虫间接飞翔肌的结构和生理特性有许多不同于脊椎动物骨胳肌的特点。蜜蜂飞翔肌肌原纤维虽然也包含有粗、细两     

甲壳动物横纹肌的收缩蛋白质与调节机制

甲壳动物横纹肌肌原纤维的肌丝陈列,收缩蛋白质和收缩的Ca²⁺依赖性调节机制与脊椎动物横纹肌有不少差异.脊椎动物横纹肌、甲壳动物快肌与慢肌的粗丝与细丝的数量比依次为1:2,1:3和1:6,肌丝阵列各异.甲壳动物粗肌丝由肌球蛋白和副肌球蛋白组成,其分子装配与脊椎动物不同.细肌丝含有肌动蛋白、原肌球蛋白和肌钙蛋白,肌钙蛋白-T分子量较高,肌钙蛋白-C仅1个Ca²⁺结合位点.甲壳动物横纹肌兼有细肌丝调节与粗肌丝调节.     

鸣鸣蝉（Oncotympana maculaticollis Motsch）发声肌中肌原纤维的双阵列结构及莫发声原初过程的分析

给出了鸣鸣蝉发声肌肌原纤维的双阵列结构,其肌纤维中并存两种不同阵列的“快”和“慢”动肌原纤维（ＦＳＭ和ＳＳＭ）．ＦＳＭ和ＳＳＭ虽然由粗肌丝构成相同的阵列骨架,但细肌丝对粗肌丝的比例（ＲＴＩＦ）不同,分别为３：１和５：１．明显区别于单音调鸣声的蝉类发声肌肌原纤维的ＲＴＩＦ为３：１的单阵列结构,即与鸣鸣蝉变音调声产生的原初机制相适应．     

鸣鸣蝉发声肌的结构观察

鸣鸣蝉Onvotympana maculaticollit Motsch的发声肌平均含193个初级肌束,多数初级肌束含9-10条肌纤维,其顶、底瑞的附着结构仅由柱状粘和细胞层组成。每条肌纤维约含1 900根肌原纤维,多数肌原纤维的长,宽和截面分别约0.77μm、0.68μm和0.53μm².井约含200根粗肌丝,其粗细肌丝的比值一般为3∶1。肌小节的长度和z线的宽度分别约3μm 和0.2μm.三联管分别位于距两端z线约0.75μm处。肌原纤维、线粒体和微气管-肌质网的面积系数分别约31.3%、46.O%和11.9%。肌小节中粗肌丝纵贯两端z线,中间无1带;细肌丝由z线相向延伸到肌小节中央,其空区约0.15-0.25μm,并无M线。这些结构特征不仅使发声肌能够利用有限的几何空间产生最大的张力,并可适应高速串的收缩运动。     

凝溶蛋白对骨骼肌天然细肌丝的作用

凝溶蛋白是Ｆ－肌动蛋白丝的钙依赖性切割性蛋白质．经过焦磷酸溶液选择性抽提和微酸性介质的有效分离,可以得到纯度较高的天然细肌丝．在Ｃａ２＋存在时,凝溶蛋白可以切割天然细肌丝．但是,凝溶蛋白对天然细肌丝的作用时程与其对Ｆ－肌动蛋白丝的作用有着显著差异,提示细肌丝中的非肌动蛋白蛋白质可能影响了凝溶蛋白对天然细肌丝的结合或者切割速率．     

横纹肌肌原纤维的第三肌丝──肌联蛋白

实验研究证明,在动物横纹肌肌原纤维中,除包含有粗肌丝、细肌丝外,还有纤肌丝的存在,肌联蛋白（肌巨蛋白）是具有挠性的线状蛋白质,分子量为3000 000,长度约为0.9μm,跨越肌原纤维的M－线和Z－线,形成纤肌丝.其生理功能是在粗肌丝装配中具有分子模板作用,并将粗肌丝稳定于肌原纤维肌小节中央以及可参与肌球蛋白活性的调节.     

栉江珧平滑闭壳肌收缩装置的超微结构

利用电子显微镜观察了栉江珧平滑闭壳肌收缩装置的精细结构,它含有粗肌丝、细肌丝和致密体。分离的天然粗肌丝含肌球蛋白和副肌球蛋白,呈现带状和Bear-Selby网格状图象,其周期为14.5nm和7.2nm。以不含ATP的低离子强度溶液处理粗肌丝,则其近侧集聚大量细肌丝;而以微酸性、中等离子强度溶液处理粗肌丝,可以溶去肌球蛋白,但不破坏其周期性结构。以2M脲处理粗肌丝,它纵向分散成直径约有10nm的长带。     

Mechanical properties of frog muscle fibres at rest and during twitch contraction.

Data reported in the literature suggest that crossbridges in rapid equilibrium between attached and detached states (weakly binding bridges), demonstrated in relaxed skinned fibres at low ionic strength, could be present also in intact fibres under physiological conditions. In addition, it was suggested that the well known leading of stiffness over force during the tension development in stimulated muscle fibres could be due to an increased number of weakly binding bridges induced by the stimulation. The experiments reviewed in this paper were made to investigate these possibilities. Fast ramp length changes were applied to single frog muscle fibres at rest and during the early phases of activation. The corresponding force changes were analysed, searching for the components expected from the presence of weakly binding bridges. The results showed no mechanical indication for the presence of weakly binding bridges in both skinned and intact fibres, either at rest or during activation. It was also found that a portion of the fibre stiffness increase induced by stimulation leads the formation of crossbridges.     

电场对完整青蛙横纹肌收缩机理的椭圆偏振法研究

利用能反映肌纤维内部结构的椭圆偏振参数研究恒定电场,脉冲电场对完整的青蛙横纹肌收缩的影响,研究表明,电场对肌纤维作用后,肌质网释放钙离子,致使肌纤维中蛋白质分子运动后出现收缩效应。实验发现：椭圆偏参数的测量灵敏度和肌小节长度有关,蛋白质运动规律和电场的性质的关系无的差异,只有量的不同。它们的共同特点是,随着电场强度的逐渐增强,其退偏率的变化呈先下降一上升的凹形曲线,这是由于电场引起的肌球蛋白分子极     

Polarization states of diffracted light. Changes accompanying fiber activation.

Measurement of the state of optical polarization of light diffracted from single, skinned and intact fibers of anterior tibialis muscle from Rana pipiens revealed a dependence upon rigor, activation, and sarcomere length (SL) change. Changes in total birefringence, delta nT, and differential field ratio value, rT, were determined. In a relaxed, skinned fiber the total birefringence value, delta nT, decreases as sarcomere length is increased from 2.1 microns to approximately 2.8-3.0 microns. From there it increases significantly to a value of approximately 1.8 x 10(-3) at a sarcomere length of 3.6 microns. The differential field ratio, rT, also shows a biphasic response to increasing sarcomere length, first exhibiting a rapid decrease over shorter SL and leveling out after the SL is beyond 3.0 microns. In comparison, relaxed intact fibers change substantially less upon sarcomere length change, showing little change in birefringence and a small bi-phasic change in rT. Skinned fibers were activated using a solution that has the same ionic strength as the relaxing solution and allows repeatable, and sustained activation. A decrease in both delta nT and rT was observed upon fiber activation. The decrease in delta nT and rT was slightly larger at shorter sarcomere lengths than at longer lengths. Relaxed fibers placed in rigor showed changes in delta nT and rT similar to those observed in activated fibers. These results are consistent with the hypothesis that, after activation, a significant portion of the thick filament cross-bridges rotate towards the actin filament resulting in redistribution of the interfilament mass content. They are also consistent with an average orientation of crossbridges in the overlap region different from that in the nonoverlap region.     

Formation of ATP-insensitive weakly-binding crossbridges in single rabbit psoas fibers by treatment with phenylmaleimide or para-phenylenedimaleimide.

Chaen et al. (1986. J. Biol. Chem. 261:13632-13636) showed that treatment of relaxed single muscle fibers with para-phenylenedimaleimide (pPDM) results in inhibition of a fiber's ability to generate active force and a diminished ATPase activity. They postulated that the inhibition of force production was due to pPDM's ability to prevent crossbridges from participating in the normal ATP hydrolysis cycle. We find that the crossbridges produced by pPDM treatment of relaxed muscle cannot bind strongly to the actin filaments in rigor, but do bind weakly to the actin filaments in the presence and also absence of ATP. After pPDM treatment, fiber stiffness, as measured using ramp stretches of varying duration, is ATP-insensitive and identical to that of untreated relaxed fibers (both at high [165 mM] and low [40 mM] ionic strength). These results suggest that the pPDM-treated crossbridges, in both the presence and absence of ATP, are locked in a state that resembles the weakly-binding myosin ATP state of normal crossbridges. Their resemblance to the ATP-crossbridges of relaxed untreated fibers is quite strong; both bind to actin about equally tightly and have similar attachment and detachment rate constants. We also found that crossbridges are locked in a weakly-binding state after treatment with N-phenylmaleimide (NPM). In muscle fibers, this method of producing weakly-binding crossbridges appears preferable to pPDM treatment because, unlike treatment with pPDM, it does not increase the fiber's resting tension and stiffness and it does not disrupt the titin band seen on SDS-PAGE.     

The Myofilament Lattice: Studies on Isolated Fibers : III. The effect of myofilament spacing upon tension

The effect of ionic strength on the generation of tension and upon the interfilament spacing in living intact and skinned single striated muscle fibers from the walking leg of crayfish (Orconectes) were determined by isometric contraction studies correlated with low-angle X-ray diffraction. Sarcomere lengths were determined by light diffraction. Tensions were induced in intact fibers by caffeine in the bathing medium and by ionophoretic microinjection of calcium. Tensions were induced in skinned fibers by a buffered calcium-EGTA solution. The interfilament spacing of intact and skinned fibers over the range of ionic strengths investigated were determined by X-ray diffraction and correlated with the physiological data. It is demonstrated that the ionic strength affects the tension-generating capacity of the muscle as it affects the chemo-mechanical transform of excitation-contraction coupling. It is further demonstrated that interfilament spacing changes encountered during shortening and with variation in the osmotic strength have no effect upon the tension-generating capacity of muscle.     

Optical depolarization changes in single, skinned muscle fibers. Evidence for cross-bridge involvement.

Optical ellipsometry studies of single, skinned muscle fibers conducted on the diffraction orders have yielded spectra that are sensitive to the state of the fiber. The linearly polarized light field vector becomes elliptically polarized as it passes through the fiber and may be collected at the diffraction orders. Fibers that have been subjected to extraction of myosin (0.6 M KCl) retain a weak diffraction pattern and exhibit a substantially decreased depolarization of incident linearly polarized light. A significant decrease in polarization is seen in skinned fibers that are subject to an increase in pH from 7.0 to 8.0. This increase in pH results in a decrease of approximately 30% in the depolarization angle of single fibers. The major decrease in depolarization angle that we observe at pH 8.0 is consistent with the notion that as cross-bridges move out from the shaft of the thick filament, their ability to cause depolarization of the incident linearly polarized light decreases. This interpretation is also consistent with the work of Ueno and Harrington where the decrease in the ability to cross-link S-1 and S-2 to the thick filament at pH 8.2 suggests cross-bridge movement away from the thick filament. A large decrease in birefringence, seen after treatment of skinned fibers with alpha-chymotrypsin, appears to be related to the breakdown of myosin into rod, S-1, heavy meromyosin, and light meromyosin.     

Muscular contraction and cytoplasmic streaming: A new general hypothesis

Several experiments point out that some crossbridges remain attached to the thin filaments at rest. It is assumed, in this paper, that these cross-bridges exert mechanical tractions on the thin filaments, directed from the thin to the thick filaments. When contraction is triggered off, a conformational change of the attached crossbridges is induced by the chemical energy released from ATP splitting. This conformational change leads to the reduction of the mechanical tensions. The electrostatic repulsive forces between the filaments become therefore automatically preponderant. This phenomenon induces a sideways expansion of the filament lattice and, taking into account the elasticity of muscle, a contraction in the direction of the filaments. This model accounts for the most important physiological and thermodynamical properties of muscle (tension-length curves, responses to quick stretch and quick release, Fenn effect, Hill's relation, behaviour of skinned fibres). It is directly applicable to all kinds of muscles and to cytoplasmic streaming, provided only actin, but not necessarily myosin, filaments are present in the cell.     

Generation of tension by skinned fibers and intact skeletal muscles from desmin-deficient mice

We have investigated the physiological role of desmin in skeletal muscle by measuring isometric tension generated in skinned fibres and intact skeletal muscles from desmin knock-out (DES-KO) mice. About 80% of skinned single extensor digitorum longus (EDL) fibres from adult DES-KO mice generated tensions close to that of wild-type (WT) controls. Weights and maximum tensions of intact EDL but not of soleus (SOL) muscles were lowered in DES-KO mice. Repeated contractions with stretch did not affect subsequent isometric tension in EDL muscles of DES-KO mice. Tension during high frequency fatigue (HFF) declined faster and this deficiency was compensated in DES-KO EDL muscles by 5 mM caffeine which had no influence on HFF in WT EDL. Furthermore, caffeine evoked twitch potentiation was higher in DES-KO than in WT muscles. We conclude that desmin is not essential for acute tensile strength but rather for optimal activation of intact myofibres during E-C coupling.     

Effects of ruthenium red on excitation and contraction in muscle fibres with Ca2+ electrogenesis.

The effect of ruthenium red (RR) on the electrical and contractile responses, membrane Ca currents, staining patterns of the external and internal membrane system were tested in intact and mechanically skinned muscle fibres of the crayfish Astacus fluviatilis. The following results were obtained: 1. Depression of the contractile responses following membrane depolarization (twitch, tetanus, potassium contractures). 2. Caffeine contractures were unaffected in intact (100 mumol/l - 1 mmol/l RR) and blocked in skinned fibres (30 mumol/l RR). 3. Mechanical threshold and mechanical latency were increased and/or prolonged. 4. The rate of depolarization of the action potentials (AP) was decreased and decremental spread of AP was recorded. 5. Both fast and slowly inactivating Ca ionic currents were decreased and the time constants of activation (tau(m] and inactivation (tau(h] were prolonged after RR (100 mumol/l) pretreatment. 6. The penetration of RR into the T-system was inversely related to its binding to the sarcolemma. The depression of depolarization-induced contractions was most pronounced in fibres with unstained sarcolemma and stained T-tubules. In intact fibres, neither terminal cisternae nor other elements of SR were stained. On the contrary, all internal membrane structures were stained in skinned fibres. There was a gradient of staining intensity from surface toward the interior.     

Further studies on the influence of ions on the muscle fibre resting potential in the cricket Acheta domesticus

Experiments were carried out on tergosternal muscle fibres of the house cricket Acheta domesticus. Conventional microelectrode methods were used. All the investigated fibres depolarized with time (0, 60, 120 min) in statistically significant manner. Replacing CL- in the physiological solution by carbonate prevents this depolarization. Citrate anions added to the bath depolarize the fibers. Acetate does not change the resting potential. Increasing osmolarity by adding 100 or 200 mM saccharose exerts no significant influence on the level of the resting potential. Lowering the level of sodium ions leads to a slight but significant depolarization. The present work confirmed the opinion that skeletal muscle fibers of Acheta domesticus are at rest multiionic electrodes.