THE ULTRASTRUCTURE OF MAMMALIAN CARDIAC MUSCLE

Papillary muscles of rat and dog hearts were fixed in such a way as to prevent excessive shortening during the procedure. The material was embedded in either araldite or methacrylate and was stained in various ways. The filamentous fine structure of mammalian cardiac muscle is similar to that previously described for striated skeletal muscle. The sarcomeres are composed of a set of thick and thin filaments which interdigitate in the A band proper. The filament ratios and the filamentous array are in accord with those found in skeletal muscle. The functional significance of this twofold array of filaments is not entirely clear. Various other structural aspects of cardiac cells such as surface membranes, mitochondria, nuclei, and cytoplasmic granules are described. The sarcoplasmic reticulum is discussed in detail as are the various structural components forming the intercalated discs. Fairly frequent deep invaginations of the sarcolemma with basement membrane are observed in addition to the intercalated discs. These surface membrane invaginations probably explain the branching appearance of cardiac fibers as seen under the light microscope.     

SYNTHETIC STRANDS OF CARDIAC MUSCLE : Formation and Ultrastructure

Spontaneously active bundles of cardiac muscle (synthetic strands) were prepared from isolated cells of 11–13-day old embryonic chick hearts which were disaggregated with trypsin. Linear orientation of the cells was obtained by plating them on agar-coated culture dishes in which either grooves were cut in the agar film or a thin line of palladium was deposited over the agar. The influence of cell-to-cell and cell-to-substrate interactions was observed with time lapse cinematography and the formation of the synthetic strand was shown to involve both random and guided cell movements, enlargement of aggregates by accretion and coalescence, and the compact linear arrangement of cells along paths of preferential adhesion. Electron microscope investigations of these strands showed that a dispersed population of heart cells organized into an inner core of muscle cells and an outer sheath of fibroblast-like cells. The muscle cells contained well-developed, but widely spaced myofibrils, a developing sarcoplasmic reticulum associated in part with the myofibrils and in part with the sarcolemma, an abundance of nonmembrane bound ribosomes and glycogen, and a prominent Golgi complex. Numerous specialized contacts were observed between the muscle cells in the strand, e.g., fasciae adherentes, desmosomes, and nexuses. A distinct type of muscle cell characterized by its pale appearance was regularly observed in the strand and was noted to be similar to Purkinje cells described in the adult avian conduction system and in developing chick myocardium. The present findings were compared with other observations of the developing myocardium, in situ, and it was concluded that, by a number or criteria, the muscle cells of the strand were differentiating normally and suitably organized for electrophysiological studies.     

CARDIAC MUSCLE OF THE HORSESHOE CRAB, LIMULUS POLYPHEMUS : I. Ultrastructure

The fine structure of the cardiac muscle of the horseshoe crab, Limulus polyphemus, has been studied with respect to the organization of its contractile material, and the structure of its organelles and the cell junctions. Longitudinal sections show long sarcomeres (5.37 µ at L_max), wide A bands (2.7 µ), irregular Z lines, no M line, and no apparent H zone. Transverse sections through the S zone of the A band show that each thick filament is ca. 180 A in diameter, is circular in profile with a center of low density, and is surrounded by an orbit of 9–12 thin filaments, each 60 A in diameter. Thick filaments are confined to the A band: thin filaments originate at the Z band, extend through the I band, and pass into the A band between the thick filaments. The sarcolemmal surface area is increased significantly by intercellular clefts. Extending into the fiber from these clefts and from the sarcolemma, T tubules pass into the fiber at the A-I level. Each fibril is enveloped by a profuse membranous covering of sarcoplasmic reticulum (SR). Sacculations of the SR occur at the A-I boundary where they make diadic contact with longitudinal branches of the T system. These branches also extend toward the Z, enlarge at the Z line, and pass into the next sarcomere. Infrequently noted were intercalated discs possessing terminal insertion and desmosome modifications, but lacking close junctions (fasciae occludentes). These structural details are compared with those of mammalian cardiac and invertebrate muscles.     

A MORPHOMETRIC STUDY ON THE NEXUS OF RAT CARDIAC MUSCLE

A morphometric study of the nexus of rat cardiac muscle was carried out. The nexus surface of one intercalated disk of one 15 µm thick fiber is found to range between 47 µm² and 94 µm², the latter value taking into account the maximal underestimation caused by tangential sectioning. Dividing the lower, minimal value by the surface of the observed subunits (90 Å periodicity), one obtains for one intercalated disk 6.7 x 10⁵ subunits, each of them assumed to be permeated by a central pore. These pores are thought to be equivalent to intercellular channels in a recently proposed model. Taking our morphometric and recently reported physiological values, this model is examined for its consistency with a low resistance pathway between cardiac muscle cells.     

大鼠离体心肌粘性劲度定量评定的研究

本实验依据心肌工作模型和心肌顺应性的理论,从大鼠离体心肌静态、动态应力-应变指数曲线和应力松弛对数曲线中,求得反映心肌粘性劲度的指标η_1和η_2,并对心肌组织粘性成分的粘性劲度进行了定量评定。η_1和η_2值与心肌粘性劲度呈同向性变化、且重复性好。本工作提示,η_1和η_2可作为实用可靠的定量评定心肌顺应性的指标。     

肌苷对缺氧心肌跨膜电位和收缩强度的影响

本工作在正常和缺氧情况下,观察肌苷对豚鼠心室乳头肌跨膜电位和收缩强度的影响。结果表明肌苷使正常心肌细胞动作电位时间(APD_(10)、APD_(50)延长。在缺氧心肌,肌苷使细胞静息电位增大,动作电位去极化幅度增高,零期最大去极化速度加快和动作电位时间延长。肌苷增加正常心肌收缩力,使缺氧心肌收缩的衰减显著缓和,亦即使收缩功能改善,且表现剂量-依从性。肌苷对心肌细胞跨膜电位的影响提示它很可能有抗心律失常作用,特别是在缺氧心脏。肌苷对离休乳头肌收缩的影响,证明其对心肌有直接的强心作用。     

垂体后叶素和加压素对离体心肌的直接作用

本实验采用大鼠离体右心房和右心室肌条模型,观察了垂体后叶素和加压素对右心房和右心室肌的直接作用。结果表明:垂体后叶素对右心房的自主性收缩频率和幅度及右心室肌的收缩幅度均有剂量依赖性抑制作用;加压素对右心房和右心室肌收缩幅度也有剂量依赖性抑制作用,但对右心房自主节律无影响;催产素对右心房的收缩频率和幅度则均无影响。加压素V_1、V_2受体拮抗剂d(CH_2)_5Tyr(Me)AVP和d(CH_2)_5(D-Ile~2,Ile~4,Ala(NH_2)~9)AVP对垂体后叶素的负性变力作用具有不同程度的阻断作用,但对垂体后叶素的负性变时作用无阻断作用。以上结果提示,垂体后叶素的负性变力作用主要是由加压素产生的,加压素对心肌有直接的负性变力作用;垂体后叶素的负性变时作用可能是非加压素和催产素成分的作用结果。     

MITOCHONDRIA IN CARDIAC MUSCLE CELLS OF THE CANARY AND SOME OTHER BIRDS

The fine structure of mitochondria from the ventricular myocardium of canaries, sparrows, zebra finches, quail, and geese has been studied. The first three of these birds have very fast heart rates, the quail being intermediate, and the goose has a relatively slow rate. The canary heart has a unique form of mitochondrion containing large, parallel arrays of zigzag or angled cristae. Other cristae, continuous with the zigzag ones and also occupying large parts of the mitochondrial volume, are named retiform because of the hexagonal network which they form, sometimes in a single plane and sometimes three dimensional. These two types of cristae appear to be interconnectible. It is possible that there is a direct functional significance in these peculiar forms, but, in any case, the relative constancy of dimensions in these arrays is probably related to specific properties of the molecules of which the cristal membrane is composed. It is also demonstrated that this membrane is composed in part of approximately 30-A particles which are believed to be protein molecules. This unusual mitochondrial morphology is not seen either in the other fast bird hearts or in the slower ones, so that there is neither a simple correlation with heart rate nor probably with the separate parts of the cardiac cycle. Although none of the other four hearts shows more than an occasional angled crista, there does seem to be a rather gross correlation between heart rate and mitochondrial size and complexity of crista structure, but no correlation with presence or absence of zigzag forms. The cristae of quail heart mitochondria are disposed in unusually large close-packed whorls.     

THE ULTRASTRUCTURE OF FROG VENTRICULAR CARDIAC MUSCLE AND ITS RELATIONSHIP TO MECHANISMS OF EXCITATION-CONTRACTION COUPLING

Frog ventricular cardiac muscle has structural features which set it apart from frog and mammalian skeletal muscle and mammalian cardiac muscle. In describing these differences, our attention focused chiefly on the distribution of cellular membranes. Abundant inter cellular clefts, the absence of tranverse tubules, and the paucity of sarcotubules, together with exceedingly small cell diameters (less than 5 µ), support the suggestion that the mechanism of excitation-contraction coupling differs in these muscle cells from that now thought to be characteristic of striated muscle such as skeletal muscle and mammalian cardiac muscle. These structural dissimilarities also imply that the mechanism of relaxation in frog ventricular muscle differs from that considered typical of other striated muscles. Additional ultrastructural features of frog ventricular heart muscle include spherical electron-opaque bodies on thin filaments, inconstantly present, forming a rank across the I band about 150 mµ from the Z line, and membrane-bounded dense granules resembling neurosecretory granules. The functional significance of these features is not yet clear.     

A STRAND OF CARDIAC MUSCLE : Its Ultrastructure and the Electrophysiological Implications of Its Geometry

The structure of a small strand of rabbit heart muscle fibers (trabecula carnea), 30–80 µ in diameter, has been examined with light and electron microscopy. By establishing a correlation between the appearance of regions of close fiber contact in light and electron microscopy, the extent and distribution of regions of close apposition of fibers has been evaluated in approximately 200 µ length of a strand. The distribution of possible regions of resistive coupling between fibers has been approximated by a model system of cables. The theoretical linear electrical properties of such a system have been analyzed and the implications of the results of this analysis are discussed. Since this preparation is to be used for correlated studies of the electrical, mechanical, and cytochemical properties of cardiac muscle, a comprehensive study of the morphology of this preparation has been made. The muscle fibers in it are distinguished from those of the rabbit papillary muscle, in that they have no triads and have a kind of mitochondrion not found in papillary muscle. No evidence of a transverse tubular system was found, but junctions of cisternae of the sarcoplasmic reticulum and the sarcolemma, peripheral couplings, were present. The electrophysiological implications of the absence of transverse tubules are discussed. The cisternae of the couplings showed periodic tubular extensions toward the sarcolemma. A regularly spaced array of Z line-like material was observed, suggesting a possible mechanism for sarcomere growth.     

常咯啉,磷酸咯啶等药物对家兔心肌脂质易化钙离子转运的影响

已知心肌细胞膜微粒体脂质(CMML)能易化钙离子从水到脂相的转运。奎尼丁能抑制这一转运过程。为进一步证明其他带含氮碱性基团的心血管药物是否也能抑制CMML易化钙离子转运过程。本文选用了常咯啉(C),磷酸咯啶(P),奎尼下(Q),乌头碱(A),调微Ⅱ号(莨菪类,T),和非含氮碱性药物成脉安(V),研究了它们对这种易化钙转运过程的影响。结果表明C,P,Q和A在30μg/ml时对这一过程的抑制均超过70％。V和T在相同浓度时抑制作用分别为37.8％和微弱作用·C,P和Q的抑制作用可能是它们治疗作用的生化基础。V和T的药理机制可能与以上药物不同。     

INHIBITION OF MALIGNANT CELL PROLIFERATION BY CULTURE MEDIA CONDITIONED BY CARDIAC OR SKELETAL MUSCLE

The present work is an attempt to explain the high resistance of muscles to cancer development. We used primary cultures of rat skeletal and cardiac muscle, and examined the effect of the supernatant of these cultures (conditioned medium; CM) on proliferation of cancer cells. The results demonstrated that CM inhibited the proliferation of several types of malignant cells by more than 50%, without a significant inhibition on normal cells. Cell cycle analysis revealed that CM increased the number of cells in S and G2 phases, suggesting a cytostatic effect of CM. For defining the biological properties of the factor(s) which are present in the CM, skeletal muscle cultures were grown in chemically defined medium (serum free medium). The concentrated sample was applied to a Sephadex G-50 column and three fractions were obtained. Only one fraction showed inhibitory activity. Four protein bands were observed in this fraction, as revealed by SDS-PAGE. We suggest that some, or all of these proteins are responsible for inhibition of tumor cell replication.     

CARDIAC MUSCLE : A Comparative Study of Purkinje Fibers and Ventricular Fibers

With light and electron microscopy a comparison has been made of the morphology of ventricular (V) and Purkinje (P) fibers of the hearts of guinea pig, rabbit, cat, dog, goat, and sheep. The criteria, previously established for the rabbit heart, that V fibers are distinguished from P fibers by the respective presence and absence of transverse tubules is shown to be true for all animals studied. No evidence was found of a permanent connection between the sarcoplasmic reticulum and the extracellular space. The sarcoplasmic reticulum (SR) of V fibers formed couplings with the sarcolemma of a transverse tubule (interior coupling) and with the peripheral sarcolemma (peripheral coupling), whereas in P fibers the SR formed only peripheral couplings. The forms of the couplings were identical. The significance, with respect to excitation-contraction coupling, of the difference in the form of the couplings in cardiac versus skeletal muscle is discussed together with the electrophysiological implications of the differing geometries of bundles of P fibers from different animals.     

STUDIES ON MUSCLE DIFFERENTIATION. III. IMMUNO-HISTOCHEMICAL IDENTIFICATION OF TROPOMYOSIN IN SKELETAL AND CARDIAC MUSCLES OF DEVELOPING CHICK EMBRYO.*,**

Specific identification of tropomyosin was succeeded in chick embryos by an application of immuno-histochemical techniques with the antisera against frog skeletal muscle tropomyosin. The first appearance of tropomyosin was in the central part of cervical somites of stage 14 embryo, and the area containing tropomyosin extended dorso-ventrally in later stages. In the heart primordium, tropomyosin was first detected in the presumptive epimyocardium at stage 8 embryos and was found to be concentrated in the epimyocardium in later embryos. The stages of the first appearance of tropomyosin in somites and presumptive myocardium corresponded with those of the first appearance of thin filaments in these organ primordia reported by other investigators. Tropomyosin in myogenic cells and in muscle fibers was localized in cell membrane or in cell peripheries. It was also distributed in a striated pattern which seemed to be due to a localization of tropomyosin in the I-bands.     

咖啡因和茶碱对大鼠皂素蜕膜心肌肌钙蛋白Ca~(2 )敏感性和肌浆网Ca~(2 )释放的影响

在低浓度皂素（50μg／ml）制备的浆膜蜕变（通透性增高）而肌浆网（SR）膜无损的蜕膜心肌标本上,以其收缩的幅度作为SRCa(2 )释放的半定量指标;高浓度皂素（500μg／ml）制备的浆膜和SR膜均蜕变的蜕膜心肌标本,以其张力-PCa关系曲线以及产生50％最大张力所需的Ch(2 )浓度（PCa50）分别作为肌钙蛋白（TN）Ca(2 )敏感性的定性和定量指标。结果观察到：（1）在低浓度皂素蜕膜心肌标本上,5和10mmol／L咖啡因分别引起约89．2±12．7和142．5±17mg（n＝4,P＜0．05）张力的强直收缩,而5mmol／L茶碱未能引起明显的强直收缩;（2）在高浓度皂素蜕膜心肌标本上,5和10mmol／L咖啡因及5mmol／L茶碱均使张力-pCa关系曲线左移;PCa50较对照分别增加了0．261,0．274和0．212PCa单位（P均小于0．001）。上述结果提示：咖啡因和茶碱均能增高TNCa(2 )敏感性;此外,咖啡因尚有促使SR释放Ca(2 )的作用。     

举重训练对男子肌比力、肌纤维的影响

安排9名有一定训练的青少年运动员参加10周举重训练。每周6次,每次8小时。训练前后用电子计算机—X线断层扫描技术(CT)测量大腿肌肉横断面积,股外肌针刺活检取样测定Ⅰ、Ⅱ型肌纤维％和面积。测定大腿伸膝肌最大随意等长力量。将结果(实验前、后)与对照组(13名无训练者)和优秀举重运动员(健将3人、一级4人)进行比较。结果表明,有训练人(实验前、后和优秀)与没有训练人(对照)在肌比力上存有差别。短、长期举重训练后,肌比力与Ⅱ型肌纤维％呈正相关。短、长期举重训练不曾改变肌纤维类型分布。举重训练对Ⅱ型肌纤维有选择性作用,且这个作用不受年龄的影响。结果还提示,肌比力的变化及肌比力与肌纤维之间的关系都会受运动项目专项特点的影响。