食物在贻贝棘尾虫(原生动物,纤毛虫)细胞内的消化过程

利用免疫荧光显微术、透射电镜及电镜酶细胞化学技术对食物在腹毛类纤毛虫-- 贻贝棘尾虫细胞内的消化过程进行了追踪观察.食物在进入消化腔隙的初期,外面包被着一层膜结构,但此外层膜很快被消化而消失;尔后食物在贻贝棘尾虫体内的消化过程可分为两种方式:一种方式为直接在消化腔隙内完成,此过程同1982年Kaul et al.和Das s et al.的报道;而另一部分食物的消化过程表现为食物逐步向细胞质内突入,以致最终完全被细胞质包围形成一被膜包裹的圆泡状结构;在此过程中,细胞质边缘突出众多含有酸性磷酸酶等物质的小囊泡结构,小囊泡与食物融合,其内的酸性磷酸酶等水解酶释放出来, 食物的表膜逐渐瓦解,个体逐渐变小,最终消化后的营养物质被释放到细胞的消化腔隙内; 这种食物通过胞口与胞咽达消化腔隙后,再由细胞质将其包裹形成的食物泡,我们称之为" 后成食物泡".这两种消化方式与贻贝棘尾虫的消化胞器为一腔隙结构相适应[动物学报 5 4(3):510-516,2008].     

魏氏拟尾柱虫休眠包囊及其细胞器超微结构的观察

为研究纤毛虫休眠状态下细胞的分化及其胞器的特征,本文以透射电镜术显示,魏氏拟尾柱虫（Ｐａｒａｕｒｏｓｔｙｌａｗｅｉｓｓｅｉ）休眠包囊中,颗粒层壁内有小泡,表膜位置偶见小泡样结构,大部分线粒体以多个相互聚集在一起,自噬泡将线粒体等胞器包裹在内经历消化过程,细胞内膜系统十分发达。作者推测,颗粒层及表厝小泡可能是休眠细胞经由表膜进行物质交换的结构,自噬泡消化现象可能是细胞中物质和能量来源的主要途径。     

魏氏拟尾柱虫休眠包囊细胞器的电镜酶细胞化学研究

为研究纤毛虫在休眠状态下细胞生命活动的特征及细胞器的功能,应用电镜酶细胞化学方法显示,魏氏拟尾柱虫（Ｐａｒａｕｒｏｓｔｙｌａ ｗｅｉｓｓｅｉ）休眠包囊中,在自噬泡位置,酸性磷酸酶反应颗粒形成多种形态的泡状体,ＡＴＰ酶反应颗粒聚集形成泡状体或球状体,在分散分嘏的线粒体中于内膜位置形成少数琥珀酸脱氢酶反应颗粒。由所得结果推测,纤毛虫休眠包囊中的自噬泡消化现象是细胞在特殊生理条件下的重要生命活动,是休眠     

棘尾虫酸性磷酸酶的定位及诱导表达

本文利用电镜对贻贝棘尾虫(Stylonychia mytilus)体内酸性磷酸酶的定位进行了观察。发现其体内除含有溶酶体性的酸性磷酸酶(ACPase)外，在纤毛内部还存在非溶酶体性的ACPase。另外，本文还利用光镜和酶细胞化学技术观察并比较了棘尾虫年轻态和衰老态个体体内ACPase在食物诱导下的表达量随时间发生的动态变化和差异，印证了棘尾虫个体的衰老也是与细胞内的酶的活性和含量的降低相伴随的[动物学报49(2)：218—223，2003]。     

旋唇纲系统发育基因组学分析支持瘦尾虫属归于散毛目

根据传统形态学特征的系统分类学研究,将瘦尾虫属(Uroleptus)归入尾柱目(Urostylida).但是,基于单基因和多基因联合的分子系统发育学分析支持瘦尾虫属与散毛目内部类群的亲缘关系更近.为进一步探讨旋唇纲(Spirotrichea)内部类群的系统发育关系以及瘦尾虫属的系统发育地位,我们对四种旋唇纲纤毛虫(澳大...     

腹毛目纤毛虫大尾柱虫腹皮层纤毛器基部微管的荧光标记

应用荧光紫杉醇直接荧光标记,显示腹毛目纤毛虫大尾柱虫Urostyla grandis腹皮层纤毛器微管胞器由口围带、波动膜、额腹横棘毛和左、右缘棘毛等纤毛器微管、纤毛器基部附属微管等组成.其中,口围带小膜托架及其相联系的肋壁微管和波动膜基体托架,额棘毛基部前纵微管束、后纵微管束及横棘毛基部前纵微管束,中腹棘毛及左、右缘棘毛基部前纵微管束、后纵微管束和横微管束,是该纤毛虫皮层纤毛器基部的主要附属微管.据结果推测,尽管腹毛目纤毛虫的纤毛器基部微管具有相同的结构成分,但其结构的组成、分化特征、定位和定向、发达程度等均有差异.所得结果为进一步说明纤毛虫细胞皮层纤毛器的形态及其微管建构的多样性提供了新的证据资料.     

一种用于观察纤毛虫表膜下结构的扫描电镜样品制备新方法

该实验摸索出通过扫描电镜观察纤毛虫表膜下三维结构的新方法:用适当浓度的KMnO_4作为固定剂,固定虫体细胞表膜,调整固定液的渗透压使细胞在低渗溶液中胀破、细胞质溶出,表膜剥落下来、内外翻转,经脱水、冷冻干燥、喷金后,在扫描电镜下对爽口虫(Climacostomumsp.)、尾草履虫(Paramecium caudatum)及拟尾柱虫(Paraurostyla weissei)的表膜下结构进行了观察。结果表明:利用此方法能够观察到表膜下层次分明而又清晰的三维立体构象。此方法可为纤毛虫表膜及其它细胞质膜的研究提供可借鉴的样品制备新方法。     

魏氏拟尾柱虫腹皮层纤毛器微管胞器的形态及形态发生

应用荧光紫杉醇直接荧光标记和抗α-微管蛋白抗体免疫荧光标记显示,魏氏拟尾柱虫(Paraurostyla weissei)腹面皮层纤毛器微管胞器由口围带、波动膜、额腹横棘毛和左右缘棘毛等纤毛器微管、纤毛器基部附属微管等组成。其中口围带基部微管包括小膜托架、小膜附属微管;额腹横棘毛和左右缘棘毛基部附属微管包括前纵微管束、后纵微管束和横微管束,它们由各自的纤毛器基部向皮层细胞质不同方向发射,形成腹皮层表面下微管网。结果表明,魏氏拟尾柱虫的纤毛器骨架、纤毛器附属结构也是一类以微管蛋白为基本成分的微管胞器,其中缘棘毛基部附属微管具有不同于其他纤毛虫(例如棘尾虫)中所观察到的同种微管胞器的建构特征。形态发生中,前仔虫口围带在老结构位置形成,其结构建成与部分老口围带的更新有关;老缘棘毛的结构物质对新的左、右缘棘毛的发生可能具有定位作用及物质贡献,但此后新的左、右缘棘毛列分别在老缘棘毛的右侧形成,而并非是在老缘棘毛位置分化的。在有些细胞中,新的左缘棘毛左侧另有一列棘毛,这可能是形态发生中老的左缘棘毛退化不完全产生的。     

新伪尾柱虫腹皮层纤毛器微管胞器的形态和形态发生

应用荧光紫杉醇直接荧光标记法显示,腹毛目纤毛虫新伪尾柱虫(Pseudourostyla nova)腹皮层纤毛器微管胞器由口围带、波动膜、额腹横棘毛和左右缘棘毛等纤毛器微管及纤毛器基部附属微管组成.口围带基部含小膜托架及与托架相联系的肋壁微管,其中领部小膜托架间由"Λ"形微管相联接;额腹横棘毛基部含前纵微管束、后纵微管束、横微管束和周围微管束,其微管在不同棘毛基部的发达程度不一;缘棘毛基部含前纵微管束、后纵微管束.同时,对新伪尾柱虫纤毛器微管胞器的形态发生和生理改组过程进行了详细的追踪研究,并对细胞皮层的额腹棘毛定位及组成特征进行了补充报道.此外,发现形态发生末期新纤毛器微管形成时,残存部分老额棘毛、横棘毛和缘棘毛,此后老结构逐渐被吸收.结果表明,新伪尾柱虫的纤毛器基部微管具有其种的特异性,新纤毛器微管分化过程中老结构可能具有定位和物质贡献作用.     

纤毛虫念珠异列虫射出胞器的超微结构观察

应用扫描电镜术和透射电镜术显示,纤毛虫念珠异列虫(Anteholosticha monilata)的射出胞器早期发生在细胞质深处,附近有不同类型的囊泡结构。成熟后射出胞器向表膜迁移,结构由不同电子密度片层的体部、结晶状的中心轴杆部和多层膜的帽部组成。受外界刺激时胞器冲破皮层射出,形态呈"蘑菇"状。据上述观察结果推测:该射出胞器具有防御作用,它可能起源于高尔基体活动产生的小泡;在亲缘关系较近的纤毛虫中,其射出胞器可能具有相似的分化特征。     

冠突伪尾柱虫营养期和形成包囊期间细胞的超微结构

冠突伪尾柱虫营养细胞中含有轴杆样结构。细胞形成包囊期间,胞质内发生自噬作用,并产生由这聚集在一起组成的高尔基体,在高尔基体内其分泌物质聚集成高电子密度的嗜锇晶体。细胞分化的结果形成含膜粒层,内层壁和外层壁的“尾柱虫类包囊”。     

显示纤毛虫细胞微管骨架的两种荧光标记方法

应用直接荧光标记和免疫荧光标记显微术显示了几种原生动物纤毛虫(如尾草履虫、大尾柱虫、阔口游仆虫)的细胞微管骨架,并据结果提出了用所述方法制备标本时需注意的几个方面:对游仆虫,可以忽略某些步骤;对大尾柱虫各种药品的浓度以及处理时间以较小为宜。     

Observations on the Ultrastructure of Uronema spp., Marine Scuticociliates*

SYNOPSIS. Observations of the ultrastructure of marine scuticociliatids, tentatively assigned to the genus Uronema, were made by light, transmission electron, and scanning electron microscopy. Giant, cortically oriented mitochondria filled the subpellicular, intermeridional areas, and were in close association with the epiplasm immediately under the inner alveolar sac membranes. Reconstructions of serial sections of the posterior poles of ciliates indicated that the intermeridional mitochondria could fuse at that point and the entire chondriome might at times be a single organelle. A system of tubules was observed to be intimately associated with the mitochondria in the posterior region. The tubules anastomosed and were directed posteriorly into the region of the nephridial-contractile vacuole system. The outer surfaces were coated with projections arranged in helical patterns. The system may be regarded as a fluid segregation organelle. The tripartite nature of the polar basal body complex observed by silver impregnation was confirmed by transmission electron microscopy. The 3 structures were the basal body of the caudal cilium and 2 parasomal sacs. A prominent ring around the caudal cilium was observed by scanning electron micrcscopy; it is probably responsible for the silver deposition surrounding the polar basal body complex that can be seen by light microscopy of silver-impregnated specimens. The ultrastructure of the nonmotile caudal cilium and its kinetosome was unremarkable, being like that of the motile, somatic cilia. The micronuclear and macronuclear outer membranes were continuous at several sites. Such interconnections explain the intimate physical relationship between the nuclei during interphase in many ciliates, and could be a structural basis for chemical communication between the 2 nuclear types. Within the cytoplasm surrounding the opening of the cytoproct, numerous clear vesicles were observed. Their position and appearance suggested that the cytoproct may be involved in the elimination of solutions as well as solids. Food vacuoles, cortical microtubules, lamellar vesicles, disc-shaped vesicles, mucocysts, and a contractile vacuole and its pore were also observed.     

The Fine Structure of Colpoda maupasi with Special Emphasis on Food Vacuoles*

SYNOPSIS. An electron microscope study of Colpoda maupasi Enriques, isolated from the intestine of the blue-tongued skink Tiliqua nigrolutea, showed that the fine structure of this ciliate is similar in all respects to that of free-living ciliates. The correspondence applies particularly to the structure, distribution and number of mitochondria. This organelle has a rich intramitochondrial structure in the form of microvilli; it is found close to the periphery, near the nuclear apparatus and in other parts of the cytoplasm. It was concluded that the association between Colpoda maupasi and Tiliqua nigrolutea was probably accidental and limited to the cyst stage. Thus electron microscopy confirmed a conclusion arrived at by light microscopy. The presence of numerous food vacuoles made it possible to study stages of digestion within this organelle. Four major types of food vacuole were distinguished. Type 1 food vacuoles are characterized by their large size, the presence of intact bacteria and abundance of water. In type 2 the food vacuole is deprived of water, the bacteria are pressed together and the nuclei have lost their structure. Type 3 food vacuoles contain only bacterial ghosts, cytoplasmic and nuclear material having been digested. Food vacuoles of this type are found only occasionally, suggesting their short duration. It is of interest that during this transient stage the bulk of digestion takes place. In type 4 nothing reminiscent of bacteria is found; there are only myelin figures and vesicles of different sizes. Evaginations and invagnations of the vacuolar membrane and vesicles of different size and structure inside and outside the food vacuoles of types 1, 3 and 4 suggest that extensive communication exists between the cytoplasm and the food vacuole. It seems likely that enzymes are delivered to the food vacuole and digested materials are released from the food vacuole to the cytoplasm.     

阔口尖毛虫形成包囊期间细胞超微结构的观察

阔口尖毛虫形成囊期间,细胞质内出现条带状或管产产的内质网和由不同大小的囊泡组成的包囊壁前体。并且,前体的产生与内质网有关;细胞质内发生自噬泡消化现象,这是细胞将原有结构和能量进行贮存,利用的一种重要形式;大核向细胞质突出形成阿米巴形结构,这与大核向细胞质排出部分核物质有关。     

Immunocytochemical localization of acid phosphatase in rat liver

Localization of acid phosphatase (ACPase) in rat liver was investigated by immunocytochemical techniques. Rat liver was fixed by perfusion and cut into thick tissue slices, which were embedded in Epon or Lowicryl K4M. For light microscopy (LM), semithin Epon sections were stained for the enzyme ACPase by an indirect immunoenzyme technique. For electron microscopy (EM), ultra-thin Lowicryl K4M sections were stained by a protein A-gold technique. By means of LM, granular reaction deposits were observed in hepatocytes and sinus-lining cells. Stained granules were present in the juxtanuclear cytoplasm, but they did not correspond to a typical staining pattern for the Golgi complex. EM revealed that gold particles indicating ACPase antigens were present on lysosomes and on some vesicles locating in the trans Golgi region. Endosomelike vesicles were strongly positive for the labeling. Golgi cisterna were mostly negative, but weak signals were noted in dilated sacules. The plasma membranes on the sinusoidal and bile canalicular sides were labeled by a few gold particles. The results indicate that ACPase is present in endosomes and in a restricted area of plasma membrane, as well as in the lysosomal system.     

Food plaquette digestion in the ciliated protozoan Hyalophysa chattoni

Abstract. The digestion of food plaquettes in the ciliated protozoan Hyalophysa chattoni was analyzed by light and electron microscopy. Through the use of nigrosin as a tracer for light microscopy and polystyrene microparticles for electron microscopy, we have demonstrated that food plaquettes transform to late-stage digestive vesicles. Eventually, in the phoront, some of the late-stage vesicles merge to form larger fusion vesicles, which are retained in the peripheral cytoplasm of the ensuing feeding stage. After the feeding stage settles and encysts, these vesicles are either retained by the daughter cells or are left in the divisional cyst as residual bodies. Food plaquettes, digestive vesicles, and fusion vesicles stain positively with neutral red and acridine orange, indicating an acidic pH. These results portray a unique digestive pathway in which stored, undigested material is reorganized into larger fusion vesicles as the cell prepares for additional feeding.     

Microtubules Are required for motility and positioning of vesicles and mitochondria in hyphal tip cells of Allomyces macrogynus

We have used video-enhanced light microscopy and digital image processing to characterize the intracellular motility and positioning of vesicles ( approximately 1-microm diameter) and mitochondria in growing hyphal tip cells of Allomyces macrogynus. These observations were coupled with cytoskeletal inhibitory experiments to define the roles of the microtubule and actin cytoskeletons in organelle translocation and positioning. Vesicles and mitochondria were abundant in apical and subapical hypha regions. Vesicles traveled along paths that were parallel to the longitudinal axis of the cell. Anterograde (i.e., toward the hyphal apex) and retrograde (i.e., away from the hyphal apex) movements of vesicles occurred at average rates of 4.0 and 2.2 microm/s, respectively. Bidirectional travel of vesicles along common paths was noted in the cortical cytoplasm. Mitochondria were aligned mostly parallel to the long axis of the hypha, except those extending into the hyphal apex, which were oriented toward the Spitzenk?rper. In regions of the subapical hypha mitochondria were often restricted to the cortical cytoplasm and nuclei occupied the central cytoplasmic region. Mitochondria displayed rapid anterograde movements reaching speeds of 3.0 microm/s, but primarily maintained a constant position relative to either the advancing cytoplasm or the lateral cell wall. Cytoskeletal disruption experiments showed that the positioning of mitochondria and motility of vesicles and mitochondria were microtubule-based and suggested that the actin cytoskeleton played uncertain roles.     

Promotion of gas bubble formation by ingested nuclei in the ciliate,Tetrahymena pyriformis

Cells of the ciliateTetrahymena pyriformis were suspended with carmine or graphite particles or with Halobacterium gas vesicles, all of which promote bubble formation in aqueous suspensions when tested with 10 atm and above (0.1−0.5×10⁷ Pa) (carmine and graphite) or 25 atm and above (gas vesicles) of nitrogen supersaturations. All three particles were ingested, but only the gas vesicles promoted intracellular gas bubble formation if the cells containing them were nitrogen or methane saturated in a slow stepwise fashion prior to rapid decompression. Cell rupture did not occur until gas saturation pressures greater than 25 atm were used; this suggests that the ciliate pellicle and cytoplasm cannot resist the mechanical forces of an expanding gas phase induced by decompression from between 25 and 50 atm and thus provides an estimate of the physical strength of these cellular components. The inability of the ingested carmine, graphite, and collapsed gas vesicles to induce intracellular gas bubble formation suggests that the phagocytic process somehow altered them. This procedure may thus provide a tool for the study of early events in the digestive processes of ciliates.     

Distribution of Acid Phosphatase in Paramecium caudatum: Its Relations with the Process of Digestion

SYNOPSIS. The distribution of acid phosphatase was investigated at the ultrastructural level in Paramecium caudatum. Acid phosphatase occurs in endoplasmic reticulum, Golgi apparatus, food vacuoles, autophagic vesicles, vacuolar and dense bodies. Some slight deposits are also seen in the mitochondria.
These observations point out that this hydrolase activity is related to digestive processes. The enzyme, originating from the endoplasmic reticulum and Golgi apparatus reaches the food vacuole or autophagic vesicle likely via the reticulum. The digestion of the bacteria or of the enclosed organelle gives rise to electronopaque material which is later found in dense bodies. These dense bodies are likely secondary lysosomes and it is possible that they may fuse with the young food vacuole or with autophagic vesicles.