锦橙汁囊的超微结构

用常规电镜方法观察了锦橙［Ｃｉｔｒｕｓｓｉｎｅｎｓｉｓ （Ｌ．） Ｏｓｂ．］汁囊从原始细胞到发育为一个具柄的成熟汁囊的过程中,汁囊构成细胞超微结构的变化。锦橙汁囊原始细胞及发育为球状体时的构成细胞以及柱状结构顶端的细胞都是一种典型的分生组织细胞。在细胞质中有包括线粒体、质体、内质网、核糖体等丰富的细胞器,但没有观察到高尔基体。这些分生细胞分裂一段时期后就停止活动,逐渐分化为适应贮藏功能的液泡化薄壁细胞。分生细胞开始分化时,在细胞中出现许多小液泡和高尔基体。这些小液泡逐渐地融合,同时细胞质变少,最后形成一个有中央大液泡的薄壁细胞,在紧贴细胞膜的薄薄的一层细胞质中有线粒体、质体、高尔基体以及含有许多脂滴的杂色体。但成熟果实中汁囊的薄壁细胞中几乎没有任何细胞器。     

棉阿舒囊霉培养过程中菌体超微形态的变化

用电镜研究了棉阿舒囊霉在核黄素发酵过程中超显微结构的变化。观察中可见线粒体、内质网、高尔基体、细胞核和液泡等亚细胞结构。对数生长早期的菌体中含有丰富的、崤发育良好的典型线粒体结构,之后就迅速减少。在数量下降同时,线粒体结构变得模糊不清而失去其完整性。 这些变化与本实验室早期报道的生化研究结果完全一致。对数生长期菌体内尚含有发育良好的内质网和高尔基体。这些细胞器也随着菌体的分化而迅速消失。菌体发育后期,则见到细胞核的分裂和配子的形成时期细胞核的结构。     

日本沼虾高尔基体在精子发生过程中的变化

用岸民镜技术研究了日本沼虾精子发生过程中生精细胞内高尔基体变化。结果表明：精原细胞内,高尔基体结构典型,分布在核膜附近,许多膜囊通过过连接小管相互连接。初级精母细胞内,高尔基体结构紧凑且更典型,更造近核膜,在反面的分泌活动旺盛,产生大量初级溶酶体;     

精子细胞高尔基体的冰冻蚀刻电镜观察

本实验用冰冻蚀刻方法观察大鼠精子细胞高尔基体的膜结构。冰冻蚀刻复型膜显示了高尔基体膜囊上的膜内颗粒和圆形凹陷结构,这些结构在高尔基体的各层膜囊以及GERL上的数目和分布是不同的。一般认为膜内颗粒代表镶嵌于生物膜类脂双分子层中的蛋白质和酶,因此,它们的数目和分布说明了精子细胞高尔基体各层膜囊结构的分子排列以及酶的活性是不一样的。圆形凹陷结构的具体性质还不清楚,它们可能与高尔基体膜囊融合运输小泡以及长出分泌小泡的过程有关。本文还把高尔基体各部位的膜内颗粒分布与它们的细胞化学反应联系起来进行了讨论。     

植物细胞内膜运输对植物发育的调控机制

植物细胞内膜体系与别的真核生物体系相似,由一系列连续的内膜结构组成,包括核膜、内质网、高尔基体、反式高尔基体、液泡、转运囊泡和细胞膜。其中,反式高尔基体和液泡具有植物细胞特有的结构特征和对应功能。从几个方面综述植物细胞内膜运输对植物发育的调控作用,包括内膜运输与生长素极性运输和稳态平衡之间的关系,内膜运输对根毛极性生长的作用机理,以及液泡运输在花粉管极性生长和种子萌发过程中的重要作用。旨在对致力于植物细胞内膜运输与发育调控的研究人员提供参考。     

体外小鼠胚胎性癌细胞BEC-B7建株和特性

恶性畸胎癌干细胞又叫做胚胎性癌细胞(简称EC细胞),是一种来自生殖细胞或早期胚胎细胞的恶性癌细胞,不仅具有类似于早期胚胎细胞分化多能性,而且能在一定条件下失去恶性生长性质,分化为各个胚层的正常组织。EC细胞是开展哺乳类胚胎早期发育遗传以及癌变和癌细胞分化研究较好的实验材料。EC细胞在体外可以大量培养生长,较之正常胚胎细胞更容易获取。我们在建立体内腹     

菲律宾蛤仔的精子发生和精子超微结构

用透射电镜研究了菲律宾蛤仔(Ruditapes philippinarum)精子结构和精子发生过程中细胞形态结构的变化及细胞器的演变规律。菲律宾蛤仔雄性生殖细胞的形态由椭圆形渐变为辣椒状,细胞核的形态由椭圆形逐渐拉长,渐变为锥形。染色质的凝集经历：小颗粒团块状一较大颗粒均匀状一粗颗粒均匀状的过程。线粒体在演化过程中数量先增多后逐渐减少,嵴数逐渐增多,电子密度和体积逐渐增大。高尔基体在初级精母细胞期已经发育,随后的各期中发育良好,分泌旺盛。精细胞Ⅱ期,高尔基体分泌的潴泡开始融合,形成前顶体囊。精细胞Ⅲ期,高尔基体的分泌物仍不断融合。精细胞分化的后期,前顶体囊逐渐发育形成顶体。菲律宾蛤仔成熟精子呈长辣椒状,为原生型,由头部、中段和尾部构成。头部的顶体为细长柱形,末端渐细,电子密度较小;细胞核为锥形;中段线粒体4个,尾部鞭毛为典型的“9 2”型结构。此外在成熟精子线粒体环横切面有一特殊“风车状”结构。     

囊泡转运复合物Retromer和SNX蛋白家族在发育和疾病中的作用

逆向囊泡转运复合物Retromer主要负责介导货物蛋白从内体向反式高尔基体或细胞表面逆向转运,是细胞内囊泡转运分选系统的重要成员.Retromer复合物主要含有两个亚复合体:货物选择复合体VPS26-VPS29-VPS35和膜结合复合体SNX-BAR.本文着重综述了Retromer复合物和SNX蛋白家族参与囊泡转运过程的分子机制以及它们在发育中对Wnt信号的调控作用;并讨论了Retromer复合物在细胞极性形成、细胞凋亡、神经元信号传递中的重要作用;以及该复合物与帕金森和阿尔茨海默病等退行性疾病之间的关系.     

酵母Dop1及其同源蛋白DOPEY对细胞糖基化和囊泡运输影响的研究

蛋白质糖基化是一种保守的翻译后修饰,对多种细胞现象至关重要。在酵母或动物细胞高尔基体中的糖链处理由结构相似的糖基转移酶或糖苷酶催化。囊泡运输等多种因素会影响糖基转移酶在高尔基体中的稳态定位,进而影响糖基化。该研究探讨高尔基外周蛋白Dop1对细胞糖基化和囊泡运输的影响。共聚焦荧光显微镜活细胞成像显示,Dop1主要定位于晚期高尔基体。Dop1及其相互作用蛋白Neo1(P4 ATPase)均参与高尔基体后期的囊泡运输。此外,Dop1介导糖基转移酶Och1的逆向运输而影响糖基化。进一步,哺乳动物DOPEY1和DOPEY2是酵母Dop1的同源蛋白。DOPEY1或DOPEY2的缺失导致高尔基体结构的改变,轻微地影响细胞糖基化。综上,酵母Dop1和哺乳动物DOPEY都参与了细胞后期的蛋白质囊泡运输,并影响高尔基体形态或糖基化。     

具有极性的分泌运输是神经元树突发育的形态和走向的基础

神经元在生长发育过程中会定向地生长出一根轴突和多根形态、功能不一的树突。虽然已经有一些关于轴突和树突分化的学说,但是关于树突形成时膜成分加载的细胞学机制仍不清楚。2005年11月Duke大学的Michael D．Ehlers实验室报道,神经元具有极性的分泌性运输方式是产生各种形态和方向树突的基础。在一般细胞内,高尔基体在细胞核周围呈现扁平膜囊的堆叠结构;而在神经元中,高尔基体既包括胞体的膜囊堆叠,又包括了树突内离散的高尔基体“前哨”（outpost;Golgi outposts指一些存在于树突中的高尔基体膜囊,本文译为“高尔基体前哨”）。Ehlers等的实验证明,在培养的海马锥体神经元中,后高尔基体的膜运输是朝向较长的树突的;而在体内,皮层和海马区的锥体神经元都有一根特化的向皮层表面行走的顶树突,后高尔基体的膜运输就朝向这根顶树突。他们发现,小的高尔基体“前哨”选择性地分布在较长的树突中,但不进入轴突。在树突中,高尔基体“前哨”常常集中在树突的分支处进行后高尔基体的运输。     

Changes of Golgi complex during the induced differentiation of human gastric adenocarcinoma cells in vitro

Golgi complex is a central link related to each kind of organella in cellular metabolic process, its morphological changes are often concerned with the cell differentiation and functional state. The Golgi complexes in tumour cells have the characteristics of poor development and non-typical structure, and closely related to their pathological differential degree. For this reason, having based on appraising the differential effect of human gastric adenocarcinoma cell line MGc 80-3 induced by dBcAMP in vitro, we had made a systematic observation on the changes of Golgi complex during the malignant phenotypical reversion of MGc 80-3 cells, in order to inquire into the relationship between the structural and functional changes of Golgi complex and the malignant phenotypic reversion of cancer cells. It was revealed by ultrathin sectioning and freeze-etching electron microscopy that, in MGc 80-3 cells, there were a few Golgi complexes, their volume was small, the amounts of saccule were few, they arranged irregularly, expanded and inflated, and the intramembranous particles on saccule were rare and not well-distributed. It displayed a non-developmental and non-typical structural state of Golgi complex. But after induced treatment with dBcAMP, the Golgi complexes had grown in number and distributed concentrated, , their volume enlarged, the saccules increased and arranged regularly, and the intermembranous particles on saccules were plentiful and well-distributed. They had restored a well-developed and typical structure of Golgi complex similar to that of the primary culture cell of human normal gastric mucous membrane. It showed that Golgi complex had further changed into a quite developed state during the induced differentiation. This alteration not only inhibited the malignant secretory activity of gastric carcinoma cells but also played a certain regulative role in the changes of the surface components of cancer cells. The structural and functional changes of Golgi complex were considered to be an important expression in the malignant phenotypical reversion of cancer cells, it had an important influence on the differentiation of cancer cells from malignant to normal direction.     

The differentiation of human gastric adenocarcinoma cell line MGc80-3 induced by dibutyryl cAMP in vitro

For providing some experimental basis in establishing malignant phenotypic reversed indexes of gastric carcinoma cells, human gastric adenocarcinoma cell line MGc-80-3 was induced by dBcAMP in vitro to appraise the effect of gastric carcinoma cell differentiation by chemical inducers. Under light microscope, MGc 80-3 cells, after treated with 1 mM dBcAMP, tended to be flat and disperse, and their volume gradually enlarged, with their nucleus relatively smaller and their shape rather regular. Morphological changes, became like normal differentiated epithelial cells, were observed. The cells attached firmly, grew slowly, their growth curve showed inhibitory rate amounted to 52.87%, and cellular division exponent displayed their peak value 1.5 times less than that of MGc 80-3 cells. It was clear that dBcAMP could effectively inhibit the multiplication activity of MGc-80-3 cells. In the cells after dBcAMP treatment, remarkable changes of cell surface charges was indicated by cell electrophoresis, the ratio dropped to 3.043 from 3.988, and their retardant ratio reached up to 31.2%. cAMP content in cells after this treatment, detected by cAMP and cGMP radioimmunoassay, was enhanced by 2.42 times; and cAMP/cGMP ratio, by 1.73 times. Thus, cAMP level within MGc 80-3 cells was raised obviously by dBcAMP. Heterotransplantation experiments showed that tumoriferous rate of MGc 80-3 cells (transplanted subcutaneously to BABL/c mice) amounted to 100%, and that of the cells after this treatment was only 5.6%. Their tumorigenic ability was extremely reduced. These results fully confirmed that dBcAMP was able to change MGc 80-3 cell's malignant phenotypic characteristics and produce a reversed alteration; thus, it has a remarkable inductive effect in differentiating gastric carcinoma cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

HpD-激光对离体培养人胃癌细胞杀伤效应的电镜观察

本文应用扫描和透射电镜观察了HpD-激光光敏作用对人胃癌MGc 80-3细胞超微结构的损伤效应。结果表明,光敏作用可以使多种细胞器结构受到损伤:(1) 使细胞表面微绒毛减少,出现大量泡状突起,最后质膜破裂、崩解;(2) 使线粒体肿胀、嵴被破坏,随后整个线粒体空泡化;(3) 使高尔基囊扩张,高尔基液泡和高尔基小泡形态变得极不规则;(4) 使核膜破裂,核仁崩解,染色质凝聚。不同的细胞以及不同的细胞器光敏损伤的时间和程度并不相同。本文讨论了损伤的不同步性与临床治疗不彻底和复发之间的关系。     

SYNTHESIS OF THE CARBOHYDRATE OF MUCUS IN THE GOLGI COMPLEX AS SHOWN BY ELECTRON MICROSCOPE RADIOAUTOGRAPHY OF GOBLET CELLS FROM RATS INJECTED WITH GLUCOSE-H3

It is known that colonic goblet cells utilize glucose to synthesize the carbohydrate portion of mucus glycoprotein. To determine the intracellular site of this synthesis, glucose-H³ was injected into 10-g rats. At 5, 20, 40 min, 1, 1½, and 4 hr after injection, segments of colon were fixed and prepared for electron microscope radioautography. By 5 min after injection, label had been incorporated into substances present in the flattened saccules of the Golgi complex. At 20 min, both Golgi saccules and nearby mucigen granules were labeled. By 40 min, mucigen granules carried almost all detectable radioactivity. Between 1 and 4 hr, these labeled granules migrated from the supranuclear region to the apical membrane; here, they were extruded singly, retaining their limiting membrane. The evidence indicates that the Golgi saccule is the site where complex carbohydrate is synthesized and is added to immigrant protein to form the complete glycoprotein of mucus. The Golgi saccule, distended by this material, becomes mucigen granules. It is roughly estimated that one saccule is released by each Golgi stack every 2 to 4 min: a conclusion implying continuous renewal of Golgi stacks. It appears that the Golgi synthesis, intracellular migration, and release of mucus glycoprotein occur continually throughout the life of the goblet cell.     

Lectin-labeled membrane is transferred to the Golgi complex in mouse pituitary cells in vivo

Labeling of the Golgi complex with the lectin conjugate wheat germ agglutinin-horseradish peroxidase (WGA-HRP), which binds to cell surface membrane and enters cells by adsorptive endocytosis, was analyzed in secretory cells of the anterior, intermediate, and posterior lobes of mouse pituitary gland in vivo. WGA-HRP was administered intravenously or by ventriculo-cisternal perfusion to control and salt-stressed mice; post-injection survival times were 30 min-24 hr. Peroxidase reaction product was identified within the extracellular clefts of anterior and posterior pituitary lobes through 24 hr but was absent in intermediate lobe. Endocytic vesicles, spherical endosomes, tubules, dense and multivesicular bodies, the trans-most saccule of the Golgi complex, and dense-core secretory granules attached or unattached to the trans Golgi saccule were peroxidase-positive in the different types of anterior pituitary cells and in perikarya of supraoptico-neurohypophyseal neurons; endoplasmic reticulum and the cis and intermediate Golgi saccules in the same cell types were consistently devoid of peroxidase reaction product. Dense-core secretory granules derived from cis and intermediate Golgi saccules in salt-stressed supraoptic perikarya likewise failed to exhibit peroxidase reaction product. The results suggest that in secretory cells of anterior and posterior pituitary lobes, WGA-HRP, initially internalized with cell surface membrane, is eventually conveyed to the trans-most Golgi saccule, in which the lectin conjugate and associated membrane are packaged in dense-core secretory granules for export and potential exocytosis of the tracer. Endoplasmic reticulum and the cis and intermediate Golgi saccules appear not to be involved in the endocytic/exocytic pathways of pituitary cells exposed to WGA-HRP.     

正丁酸钠对人胃腺癌细胞染色体数目和DNA含量的影响

培养的人胃腺癌MGc 80-3细胞经过正丁酸钠处理7天后,生长抑制率达50.7%,约有90%的细胞形态发生分化,其超微结构亦有显著改变。而且,在染色体数目上,超二倍体细胞由对照组的78%增加到实验组的96%,超三倍体和超四倍体细胞则分别从6%和14%下降至2%。同时应用~3H-TdR放射自显影和福尔根细胞光度法测定未标记细胞(G_1期)DNA含量,结果显示实验组比对照组降低了。而且在实验组的同一制片中,未分化细胞DNA含量平均为超六倍体值(DI=3.67和3.56),其中90%的细胞超过6C;分化细胞DNA含量则平均为近四倍体值(DI=2.03和1.99),其中近60%的细胞少于4C。两者差异统计显著,表明形态分化的人胃腺癌细胞的遗传物质含量明显减少,但这些细胞并非就是正常二倍体细胞。     

Ultrastructural localization of CMPase, TPPase, and NADPase activity in neurons, satellite cells, and Schwann cells in frog dorsal root ganglia

Sections of bullfrog dorsal root ganglia were analyzed for cytidine monophosphatase (CMPase), thiamine pyrophosphatase (TPPase), and nicotinamide adenine dinucleotide phosphatase (NADPase) activity, and the distributions of these enzymatic activities were compared with those traditionally found in other cell types (e.g., CMPase: Golgi trans-sacculotubular network; TPPase: trans-Golgi saccule(s); NADPase: intermediate Golgi saccules). In the present study, CMPase activity in neurons was localized mainly to the Golgi trans-sacculotubular network and lysosomes, but sometimes also occurred at the ends of the trans and most distal intermediate Golgi saccules. A similar distribution was found in satellite and Schwann cells. TPPase activity in neurons occurred not only in the trans-Golgi saccule but also in the trans-sacculotubular network, lysosomes, and scattered tubular elements. In satellite and Schwann cells, activity was found in both the trans saccule and trans-sacculotubular network, and substantial activity often appeared in the more distal of the intermediate saccules. NADPase activity in neurons was usually absent from the intermediate Golgi saccules and was confined to the trans-sacculotubular network and lysosomes; however, activity was sometimes also found in the intermediate and/or trans-Golgi saccules. In satellite and Schwann cells, activity appeared consistently in both the trans-sacculotubular network and intermediate saccules, as well as in lysosomes. These distributions, especially in the case of TPPase and NADPase, differ substantially from the most frequently reported localizations of the above enzymes, indicating that the Golgi complex may exhibit considerable plasticity of structure and function in different cell types.     

天然甾体皂甙化合物的抗肿瘤活性

采用MTT法,以长春新碱(VCR)为阳性对照,研究了6种从菝葜属植物中分离提取的天然甾体皂甙化合物对肝癌SMMC-7721、人宫颈癌HeLa和胃腺癌MGc80-3细胞生长的抑制作用.结果显示;6种甾体皂甙抗肿瘤活性与其化学结构密切相关,对三种癌细胞的抑瘤作用强度相同,抑癌活性的顺序为:薯蓣皂甙>VCR>SQD_4>SQD_3>M_1>SQD_1,甲基原薯蓣皂甙.甾体骨架的差异性是决定这类化合物抗肿瘤活性的主要因素.     

Polyanion-mediated mineralization — assembly and reorganization of acidic polysaccharides in the Golgi system of a coccolithophorid alga during mineral deposition

Summary Immunolocalization of two highly acidic polysaccharides (PS-1 and PS-2) in a calcifying algaPleurochrysis carterae is described throughout the mineralization process, from before crystal nucleation through the cessation of crystal growth. This unicellular coccolithophorid alga is a useful model for mineralization because it produces calcified scales known as coccoliths in homogeneous cell culture. PS-1 and PS-2 were localized in the crystal coats of mature coccoliths and in electron dense Golgi particles. The polyanions are synthesized in medial Golgi cisternae and co-aggregate with calcium ions into discrete 25 nm particles. Particle-laden vesicles bud from cisternal margins and fuse with a coccolith-forming saccule containing an organic oval-shaped scale which forms the base of the future coccolith. The particles are localized on the base before the onset of mineral deposition and are present in the coccolith saccule throughout the period of crystal (CaCO₃) nucleation and growth. During the final phase of coccolith formation, the particles disappear, and the mature crystals acquire an amorphous coat containing PS-1 and PS-2 polysaccharides which remain with the mineral phase after the coccoliths are extruded from the cell. Postulated mechanisms of polyanion-mediated mineralization are reviewed and their relevance to the calcification of coccoliths is addressed.Abbreviations PS-1 polysaccharide one - PS-2 polysaccharide two - BSA bovine serum albumin - SDS sodium dodecyl sulfate - MES 2-(N-morpholino)-ethanesulfonic acid - EDTA ethylenediaminetetraacetic acid - DHA 3-deoxy-lyxo-2-heptulosaric acid - TCA trichloroacetic acid     

DETECTION OF COMPLEX CARBOHYDRATES IN THE GOLGI APPARATUS OF RAT CELLS

Two methods used for the electron microscopic detection of glycoproteins were applied to a variety of cell types in the rat; one involved successive treatment of sections with periodic acid, chromic acid, and silver methenamine; and the other, a brief treatment with a chromic acid-phosphotungstic acid mixture. The results obtained with the two methods were identical and, whenever the comparison was possible, similar to those obtained with the periodic acid-Schiff technique of light microscopy. In secretory as well as in nonsecretory cells, parts of the Golgi apparatus are stained. The last saccule on one side of each Golgi stack is strongly reactive (mature face), and the last saccule on the other side shows little or no reactivity (immature face); a gradient of reactivity occurs in between these saccules. The more likely explanation of the increase in staining intensity is that carbohydrate is synthesized and accumulates in saccules as they migrate toward the mature face. In many secretory cells, the mature face is associated with strongly stained secretory granules. Other structures stained are: (1) small vesicles, dense and multivesicular bodies, at least some of which are presumed to be lysosomal in nature; (2) cell coat; and (3) basement membrane. The evidence suggests that the Golgi saccules provide glycoproteins not only for secretion, but also for the needs of the lysosomal system as well as for incorporation into the cell coat and perhaps basement membrane.