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The sarcotubular system of frog skeletal muscle. A morphological and biochemical study. 青蛙骨骼肌的肌管系统。形态学和生物化学的研究
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.201
U MUSCATELLO, E ANDERSSON-CEDERGREN, G F AZZONE, von der DECKEN

In the frog skeletal muscle cell a well defined and highly organized system of tubular elements is located in the sarcoplasm between the myofibrils. The sarcoplasmic component is called the sarcotubular system. By means of differential centrifugation it has been possible to isolate from the frog muscle homogenate a fraction composed of small vesicles, tubules, and particles. This fraction is without cytochrome oxidase activity, which is localized in the mitochondrial membranes. This indicates that the structural components of this fraction do not derive from the mitochondrial fragmentation, but probably from the sarcotubular system. This fraction, called sarcotubular fraction, has a Mg(++)-stimulated ATPase activity which differs from that of muscle mitochondria in that it is 3 to 4 times higher on the protein basis as compared with the mitochondrial ATPase, and is inhibited by Ca(++) and by deoxycholate like the Kielley and Meyerhof ATPase. We therefore conclude that the "granules" of the Kielley and Meyerhof ATPase, which were shown to have a relaxing effect, are fragments of the sarcotubular system. The isolated sarcotubular fraction has a high RNA content and demonstrable activity in incorporating labeled amino acids, even in the absence of added supernatant.

在青蛙的骨骼肌细胞中,在肌原纤维之间的肌质中有一个定义明确、组织严密的管状细胞系统。肌浆成分称为肌管系统。通过差速离心,可以从青蛙肌肉匀浆中分离出由小泡、小管和颗粒组成的部分。这个部分没有细胞色素氧化酶活性,它位于线粒体膜上。这表明该部分的结构成分不是来自线粒体碎片,而可能来自肌管系统。这个部分被称为肌管部分,具有Mg(++)刺激的atp酶活性,与肌肉线粒体的atp酶活性不同,因为它在蛋白质基础上比线粒体atp酶高3到4倍,并且被Ca(++)和脱氧胆酸(如Kielley和Meyerhof atp酶)抑制。因此,我们得出结论,Kielley和Meyerhof atp酶的“颗粒”具有放松作用,是肌管系统的碎片。分离的肌小管部分具有高RNA含量和可证明的结合标记氨基酸的活性,即使在没有添加上清的情况下。
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引用次数: 107
FUNCTIONAL ANALYSIS OF SWIM-BLADDER MUSCLES ENGAGED IN SOUND PRODUCTION OF THE TOADFISH. 蟾鱼参与发声的鱼鳔肌肉的功能分析。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.187
C R Skoglund

A functional analysis of the striated swim-bladder muscles engaged in the sound production of the toadfish has been performed by simultaneous recording of muscle action potentials, mechanical effects, and sound. Experiments with electrical nerve stimulation were made on excised bladder, while decerebrate preparations were used for studies of reflex activation of bladders in situ. The muscle twitch in response to a single maximal nerve volley was found to be very fast. The average contraction time was 5 msec. with a range from 3 to 8 msec., the relaxation being somewhat slower. The analysis of muscle action potentials with surface electrodes showed that the activity of the muscle fibers running transversely to the long axis of the muscle was well synchronized both during artificial and reflex activation. With inserted metal microelectrodes monophasic potentials of 0.4 msec. rise time and 1.2 to 1.5 msec. total duration were recorded. The interval between peak of action potential and onset of contraction was only 0.5 msec. Microphonic recordings of the characteristic sound effect accompanying each contraction showed a high amplitude diphasic deflection during the early part of the contraction. During relaxation a similar but smaller deflection of opposite phase could sometimes be distinguished above the noise level. The output from the microphone was interpreted as a higher order derivative function of the muscle displacement. This interpretation was supported by complementary experiments on muscle sound in mammalian muscle. The dependence of the sound effects on the rate of muscle contraction was demonstrated by changing the temperature of the preparation and, in addition, by a special series of experiments with repeated stimulation at short intervals. Results obtained by varying the pressure within the bladder provided further evidence for the view that the sound initiated in the muscle is reinforced by bladder resonance. Analysis of spontaneous grunts confirmed the finding of a predominant sound frequency of about 100 per second, which was also found in reflexly evoked grunts. During these, muscle action potentials of the same rate as the dominant sound frequency were recorded, the activity being synchronous in the muscles on both sides. Some factors possibly contributing to rapid contraction are discussed.

通过同时记录肌肉动作电位、机械效应和声音,对参与蟾蜍鱼发声的条纹鱼鳔肌肉进行了功能分析。在切除膀胱上进行神经电刺激实验,用脱脑制剂原位研究膀胱的反射激活。我们发现,对单一最大神经截击的肌肉抽搐反应非常快。平均收缩时间为5毫秒。范围从3到8毫秒。弛豫稍微慢一些。表面电极对肌肉动作电位的分析表明,在人工激活和反射激活过程中,肌肉长轴横向肌纤维的活动同步良好。插入金属微电极的单相电位为0.4毫秒。上升时间和1.2 ~ 1.5毫秒。记录总持续时间。动作电位峰值与收缩开始的时间间隔仅为0.5 msec。每次收缩所伴随的特征性声音效果的麦克风记录显示,在收缩的早期有一个高振幅的双相偏转。在弛豫期间,有时可以在噪声级以上分辨出类似但较小的相反相位的偏转。麦克风的输出被解释为肌肉位移的高阶导数函数。这一解释得到了哺乳动物肌肉声音的补充实验的支持。声音效果对肌肉收缩速率的依赖性是通过改变制备温度来证明的,此外,通过短时间间隔重复刺激的一系列特殊实验来证明。通过改变膀胱内的压力得到的结果进一步证明了由肌肉发出的声音通过膀胱共振得到加强的观点。对自发咕噜声的分析证实了一项主要的声音频率约为每秒100次的发现,这在反射性唤起的咕噜声中也发现了。在此过程中,记录了与主导声音频率相同速率的肌肉动作电位,两侧肌肉的活动是同步的。讨论了一些可能导致快速收缩的因素。
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引用次数: 172
Incorporation of C14-labeled maleic hydrazide into the root-tip cells of Allium cernuum, Vicia faba, and Tradescantia paludosa. c14标记的马来酰肼在葱、蚕豆和白草根尖细胞中的掺入。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.567
J J CALLAGHAN, P GRUN

Allium cernuum, Vicia faba, and Tradescantia paludosa were treated by root immersion in maleic hydrazide (1 mM/liter) labeled with C(14) (C(14)-MH) for 1 hour to determine the location within the cell to which MH moves during various periods of time after treatment. Root tips were fixed 24 hours, 48 hours, 72 hours, and 3 weeks after treatment. Autoradiographs of root tips squashed 24 to 72 hours after fixation showed that C(14)-MH was distributed throughout the nuclei and was particularly concentrated in the nucleoli. The nucleolar localization of the chemical was transitory, fixations made 3 weeks after treatment showing well labeled nuclei many of which completely lacked label in the nucleoli. The chromosomes seen in mitotic divisions of all three species had the same amount of label in euchromatic as heterochromatic areas. Since the chemical was not accumulated preferentially in heterochromatic areas, it seems likely that the reported specificity of MH for the breakage of heterochromatin can not be due to preferential heterochromatic incorporation.

用C(14) (C(14)-MH)标记的马来酰肼(1 mM/l)浸泡葱、蚕豆和白草根1小时,测定处理后不同时期MH在细胞内移动的位置。治疗后24小时、48小时、72小时和3周固定根尖。固定24 ~ 72小时后根尖压扁的x射线自显像显示,C(14)-MH分布于细胞核各处,尤其集中于核仁。该化学物质的核仁定位是短暂的,治疗后3周的固定显示细胞核标记良好,其中许多核仁完全没有标记。在有丝分裂中,这三个物种的染色体在正染色质区和异染色质区具有相同数量的标记。由于该化学物质并不是优先在异染色质区域积累,因此所报道的MH对异染色质断裂的特异性似乎不可能是由于优先的异染色质结合。
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引用次数: 24
Observations on the fine structure and cytochemistry of mouse and human intercostal neuromuscular junctions. 小鼠和人肋间神经肌肉连接的精细结构和细胞化学观察。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.517
S I ZACKS, J M BLUMBERG

The fine structure of the mouse and human intercostal muscle neuromuscular junction was studied after brief fixation in a new formol-sucrose fixative. This primary formalin fixation was followed by brief postosmication in buffered 1 per cent osmium tetroxide. Muscle blocks were embedded in methacrylate or Epon 812 epoxy resin. Marked similarities between mouse and human motor end-plates were observed. Neuromuscular junctions from both mouse and human intercostal muscle showed synaptic vesicles, primary and secondary synaptic clefts, and layered differentiation of the amorphous surface material (ASM) present on the surface of the Schwann cell plasma membrane and on the muscle surface membrane in the region of the neuromuscular junction. An attempt to stain the ASM with lead was unsuccessful. Observations on thick and thin plastic-embedded sections stained by PAS after diastase digestion showed that the ASM within the subneural apparatus is PAS positive. Alcian blue stained the endoneurium and perineurium of peripheral nerve bundles and portions of the end-plates. The similarity of the PAS-positive ASM to other basement membranes described in other sites is discussed and its possible physiologic significance within the subsynaptic apparatus is considered.

用新型甲醛-蔗糖固定物短暂固定小鼠和人肋间肌神经肌肉接点后,观察其精细结构。初次固定福尔马林后,在1%的四氧化锇缓冲液中进行短暂的后灌注。肌块包埋于甲基丙烯酸酯或Epon 812环氧树脂中。观察到小鼠和人类运动终板有明显的相似之处。小鼠和人肋间肌神经肌肉连接处均可见突触囊泡、原发性和继发性突触间隙,神经肌肉连接处雪旺细胞细胞膜表面和肌肉表面膜上存在分层分化的无定形表面物质(ASM)。用铅染色ASM的尝试没有成功。淀粉酶消化后PAS染色的厚、薄塑料包埋切片显示,亚神经装置内的ASM为PAS阳性。阿利新蓝染色周围神经束和部分终板的神经内膜和神经周围膜。讨论了pas阳性ASM与其他部位描述的其他基底膜的相似性,并考虑了其在亚突触内可能的生理意义。
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引用次数: 43
The development of vaccinia virus in Earle's L strain cells as examined by electron microscopy. 用电子显微镜观察厄尔氏L株细胞中牛痘病毒的发育。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.475
S DALES, L SIMINOVITCH

A favorable system which is amenable to frequent and reproducible sampling, consisting of suspension cultures of strain L cells and vaccinia virus, was employed to study the animal virus-mammalian host cell relationship. The three principal aspects investigated concerned the adsorption and penetration of vaccinia into the host, the relationship between the sequence of virus development and the production of infectious particles, and the changes in the fine structure of the host cells. Experiments in which a very high multiplicity of infection was used revealed that vaccinia is phagocytized by L cells in less than 1 hour after being added to the culture, without any apparent loss of its outer limiting membranes. Regions of dense fibrous material, thought to be foci of presumptive virus multiplication, appear in the cytoplasm 2 hours after infection. A correlation between electron microscope studies and formation of infectious particles shows that although immature forms of the virus appear 4 hours after infection, infectious particles are produced 6 hours after infection of the culture, at the time when mature forms of vaccinia appear for the first time in thinly sectioned cells. Spread of the infection is gradual until eventually, after 24 hours, virus is being elaborated throughout the cytoplasm. Addition of vaccinia to monolayer cultures induced fusion of L cells and rapid formation of multinucleate giant forms. In both suspension and stationary cultures infected cells elaborate a variety of membranous structures not present in normal L cells. These take the form of tube-like lamellar and vesicular formations, or appear as complex reticular networks or as multi-laminar membranes within degenerating mitochondria.

采用L株细胞和牛痘病毒的悬浮培养系统,研究了动物病毒与哺乳动物宿主细胞的关系。研究的三个主要方面涉及牛痘对宿主的吸附和渗透,病毒发育序列与感染性颗粒产生的关系,以及宿主细胞精细结构的变化。在高感染倍数的实验中发现,牛痘在加入培养液后不到1小时就被L细胞吞噬,没有任何明显的外限制膜损失。在感染后2小时,细胞质中出现被认为是推测病毒增殖的密集纤维物质区域。电镜研究与感染性颗粒形成之间的相关性表明,尽管未成熟的病毒形式在感染后4小时出现,但感染性颗粒在培养物感染后6小时产生,此时成熟的痘苗形式首次出现在薄切片细胞中。感染的扩散是逐渐的,直到最终,24小时后,病毒在细胞质中形成。在单层培养物中加入牛痘诱导L细胞融合并快速形成多核巨细胞。在悬浮和固定培养中,感染的细胞精心制作了正常L细胞中不存在的各种膜状结构。它们以管状片层和泡状结构的形式出现,或在退化的线粒体内以复杂的网状网络或多层膜的形式出现。
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引用次数: 279
The fine structure of neuromuscular junctions and the sarcoplasmic reticulum of extrinsic eye muscles of Fundulus heteroclitus. 异交底外源性眼肌神经肌肉连接及肌浆网的精细结构。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.111
J F REGER

The extrinsic eye muscles of the killifish (F. heteroclitus) were fixed in OSO(4) (pH 7.6) and subsequently dehydrated, embedded, and sectioned for electron microscopy. The fine structures of neuromuscular junctions and of sarcoplasmic reticulum were then observed. The neuromuscular junction consists of the apposition of axolemma (60 to 70 A) and sarcolemma (90 to 100 A), with an intervening cleft space of 200 to 300 A, forming a synaptolemma 400 to 500 A thick. The terminal axons contain synaptic vesicles, mitochondria, and agranular reticulum. The subsynaptic sarcolemma lacks the infolding arrangement characteristic of neuromuscular junctions from other vertebrate skeletal muscle, making them more nearly like that of insect neuromuscular junctions. A comparison between the folded and non-folded subsynaptic membrane types is made and discussed in terms of comparative rates of acetylcholine diffusion from the synaptic cleft and resistances of the clefts and subsynaptic membranes. The sarcoplasmic reticulum consists of segmentally arranged, membrane-limited vesicles and tubular and cisternal elements which surround individual myofibrils in a sleeve-like arrangement. Triadic differentiation occurs at or near the A-I junction. Unit sleeves span the A and I bands alternately and consist of closed terminal cisternae interconnected across the A and I bands by tubular cisternae. The thickness of the sarcoplasmic membranes increases from 30 to 40 A in intertriadic regions to 50 to 70 A at the triads. The location of the triads is compared with previously described striated muscle from Ambystoma larval myotomes, cardiac and sartorius muscles of the albino rat, mouse limb muscle, chameleon lizard muscle, and insect muscle, with reference to their possible role in intracellular impulse conduction.

将鳉鱼(F. heteroclitus)的外源眼肌固定在OSO(4) (pH 7.6)中,随后脱水、包埋并切片用于电镜观察。观察神经肌肉连接处和肌浆网的精细结构。神经肌肉连接处由轴膜(60 ~ 70a)和肌膜(90 ~ 100a)的并列组成,中间间隙为200 ~ 300a,形成400 ~ 500a厚的突触膜。末端轴突包含突触囊泡、线粒体和颗粒网。突触下肌膜缺乏其他脊椎动物骨骼肌神经肌肉连接的内折叠排列特征,使它们更接近于昆虫的神经肌肉连接。从乙酰胆碱从突触间隙扩散的比较速率和突触间隙和亚突触膜的阻力方面,对折叠和非折叠的突触亚膜类型进行了比较和讨论。肌浆网由节段排列、膜限制的囊泡、管状和池状元件组成,它们以套筒状排列围绕着单个肌原纤维。三联分化发生在A-I交界处或附近。单元套筒交替跨越A和I波段,由封闭的终端蓄水池组成,通过管状蓄水池在A和I波段相互连接。肌质膜厚度从三合一区30 ~ 40a增加到三合一区50 ~ 70a。将三合体的位置与先前描述的Ambystoma幼虫的横纹肌、白化大鼠的心肌和缝口肌、小鼠肢体肌肉、变色龙蜥蜴肌肉和昆虫肌肉的横纹肌进行比较,以参考它们在细胞内冲动传导中的可能作用。
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引用次数: 54
The sarcoplasmic reticulum. Its recent history and present status. 肌浆网。它的近代史和现状。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.219
K R PORTER
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引用次数: 333
Investigations on the fine structure of striated muscle fiber read before the Reale Istituto Lombardo, 13 March 1902. 对横纹肌纤维精细结构的研究,于1902年3月13日在伦巴多皇家研究所发表。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.1
E VERATTI
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引用次数: 88
Pore canals and related structures in insect cuticle. 昆虫角质层的孔道及相关结构。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.589
M LOCKE

The fine structure and the distribution of an esterase have been studied in the cuticle of Galleria larvae, Tenebrio larvae and pupae, and in the wax-secreting cuticle of the honey bee, and compared with those in the cuticle of the caterpillar of Calpodes. In Galleria and Tenebrio the pore canals are spaces passing through the lamellate endocuticle from the epithelium to the epicuticle. They contain a filament from the cells which may be concerned in their formation. The shape of the pore canal is probably determined by the orientation of the fibres making up the lamellae in the endocuticle and is not a regular helix. The pore canals also contain numerous filaments of another sort which pass on through the epicuticle and are believed to be the origin of the surface wax. They are particularly abundant in the pore canals of the honey bee wax-secreting cuticle and extend into the cell in long pockets surrounded by an envelope of the plasma membrane. The esterase is probably concerned with the final stage of wax synthesis, for its distribution is similar to that of the lipid filaments.

研究了一种酯酶在廊虫幼虫、黄粉虫幼虫和蛹以及蜜蜂的分泌蜡质角质层中的精细结构和分布,并与足足虫毛虫角质层中的酯酶进行了比较。在廊虫和黄粉虫中,孔管是从上皮到表皮外皮穿过层状内膜的空隙。它们含有来自细胞的细丝,这可能与它们的形成有关。孔管的形状可能是由构成内膜片的纤维的方向决定的,而不是规则的螺旋形。孔管还包含许多另一种类型的细丝,它们穿过表皮,被认为是表面蜡的起源。它们在蜜蜂分泌蜡质角质层的孔道中特别丰富,并以被质膜包围的长口袋延伸到细胞内。酯酶可能与蜡合成的最后阶段有关,因为它的分布与脂质丝的分布相似。
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引用次数: 279
Development and germination of the Azotobacter cyst. 固氮菌包囊的发育和萌发。
Pub Date : 1961-08-01 DOI: 10.1083/jcb.10.4.555
O WYSS, M G NEUMNN, M D SOCOLOFSKY

The fine structure of Azotobacter vinelandii has been studied by means of electron microscopy of ultrathin sections made of the encysting and germinating cells. The organisms were fixed with KMnO(4) and embedded in epoxy resin. On an encystment medium the rod-shaped bacteria begin to assume an almost spherical form and then bark-like exine appears in 1(1/2) to 2 days. The exine thickens and an electron permeable intine forms between it and the shrinking cell body. In 5 days the intine makes up more than half of the cyst volume and begins to show a definite two-layered structure. Meanwhile the peripheral bodies, which may be extensions of the cell membrane of the vegetative cell, disappear as the encystment progresses. The cell wall and membrane of the vegetative cell remain demonstrable as the confining structure of the shrinking central body of the mature cyst. In this central body lipoidal globules appear together with aggregations of nuclear material. Cyst germination begins with an increase in the size of the central body at the expense of the intine. The nuclear aggregations become more diffuse and the lipoidal globules disappear. The exine may be pushed outward and the bark-like fragments separate as the emerging vegetative cell develops. Invagination of the cell wall and membrane may occur at this stage leading to cell division. Empty exines remain as horseshoe-shaped structures.

本文用电子显微镜研究了葡萄色固氮菌包囊细胞和萌发细胞的超薄切片的精细结构。这些生物用KMnO(4)固定,并包埋在环氧树脂中。在包囊培养基上,杆状细菌开始呈现几乎球形,然后在1(1/2)到2天内出现树皮状的外壁。外壁增厚,在它和收缩的细胞体之间形成一个电子可渗透的内层。在5天内,囊肿的内部占囊肿体积的一半以上,并开始显示明确的双层结构。同时,外周小体,可能是营养细胞细胞膜的延伸,随着包囊的进展而消失。营养细胞的细胞壁和细胞膜仍然是成熟囊肿收缩的中心体的限制性结构。在这个中央体中,脂质球与核物质聚集在一起。囊肿萌发开始时,中心体的大小增加,而中心体的大小减少。核聚集变得更加分散,脂质球消失。随着新生营养细胞的发育,外壁可向外挤压,树皮状的碎片可分离。这一阶段细胞壁和细胞膜内陷,导致细胞分裂。空的考场仍然是马蹄形的结构。
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引用次数: 81
期刊
The Journal of Biophysical and Biochemical Cytology
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