细胞骨架与卵子减数分裂器的再启动和旋转关系的研究

本文采用体外授精(IVF)技术,通过小鼠受精卵早期发育过程中的形态变化,来研究受精过程中减数分裂器的重新启动与旋转。得到以下几个结果:①精子是在小鼠卵子的减数分裂器旋转之前进入卵子,随后精核发生去凝缩,从而使停滞的减数分裂器复苏启动。②雄性原核的形成过程,伴随着减数分裂器的旋转过程,雄性原核的形成要比雌性原核的为早。③在减数分裂器旋转过程中,微丝不仅参与,并且控制着旋转;解聚微丝,将阻止减数分裂器的旋转。④微管是减数分裂器的主要构成部分,它的稳定为减数分裂器形成提供了保证;解聚微管,将使减数分裂器解体。     

狭基巢蕨叶表皮的结构和气孔器发育的观察

狭基巢蕨Ｎｅｏｔｏｐｔｅｒｉｓａｎｔｒｏｐｈｙｏｉｄｅｓ（Ｃｈｒｉｓｔ）Ｃｈｉｎｇ叶片的上表皮无气孔器,仅具表皮细胞,下表皮由表皮细胞和气孔器组成,气孔指数为２．５。上下表皮细胞和气孔器的细胞中均含有叶绿体。每个气孔器由２个肾形的保卫细胞和２～６个副卫细胞组成,其中以３个和４个副卫细胞的占绝大多数（３细胞的占４５．１％,４细胞的占４３．５％）。从发育上看,气孔器原始细胞进行２次分裂,产生２个保卫细胞和１个同源的副卫细胞。气孔器的发育过程大体可分为４个时期：（１）气孔器原始细胞的分化和分裂期;（２）保卫细胞母细胞成熟期;（３）保卫细胞母细胞分裂和气孔器幼期;（４）气孔器成熟期。狭基巢蕨的气孔器属于中周型     

A gas-flow cryostat for use in freeze-quench studies: design and application to discontinuous pre-steady-state spectral analyses

A design for an improved freeze-quench apparatus is presented. A freeze-quench apparatus has two main parts: the apparatus which mixes the reactants and after a set time sprays the reacting mixture into the quencher, and the quenching apparatus itself. The quenching apparatus is the novel feature of our design and it comprises a gas-flow cryostat mounted directly onto a liquid nitrogen storage Dewar. The gas flow maintains the temperature of a small volume of isopentane, which acts as the freeze-quench agent, at -140 degrees C. (The apparatus can maintain the quenching temperature over the range -190 degrees C to room temperature.) Because of the small size of the cryostat and the much reduced volume of solvent used by this method it is more convenient to use than its predecessors, can be used in the open laboratory, and is safer. Our apparatus is designed for application to electron paramagnetic resonance studies but could be easily modified for use with other spectroscopic techniques.     

Sexual differences in the Golgi apparatus of rat hepatocytes: three-dimensional analysis

Lipid metabolism takes place in the Golgi apparatus, but at a higher rate in female than in male rats. I therefore examined the Golgi apparatus by morphometric means for differences between the sexes at the light-and electron-microscopic level. The Golgi apparatus was stained in situ by a zinc-iodide-osmium method. The counts of the Golgi apparatus in cross-sections in female hepatocytes by light microscopy were approximately twice that in male hepatocytes. Upon ovariectomy, these counts were greatly reduced but were reestablished after estrogen supplement. To clarify this phenomenon, three-dimensional reconstructions of the Golgi apparatus were produced from electron-microscopic images of serially cut 160-nm sections. The Golgi apparatus of both male and ovariectomized females had the shape of a small ring, whereas it took the form of a large elongated cylinder in normal females and in castrated males after treatment with estrogen. The numerical difference in Golgi apparatus counts by light microscopy of in males and females is, therefore, apparently attributable to the size and shape of the Golgi apparatus, and is controlled by the estrogen level.     

Postnatal differentiation of the Golgi apparatus and the dendrites of Purkinje cells of the rat cerebellum

Summary The morphology of postnatal differentiation of the Golgi apparatus, the nucleus, the perikaryon, and the dendrites was studied in Purkinje cells of the rat cerebellum for 30 days after birth using histochemical, histological, and electron microscopic methods.The Golgi apparatus during differentiation undergoes morphological and positional changes. From the 1st to 7th postnatal day, the Golgi apparatus is found in a supranuclear position, and is connected with the axes of differentiating primary dendrites by beam-like processes. From days 8 to 11 this connection disappears, and most of the Golgi apparatus assumes a lateronuclear and infranuclear position. After the 11th or 12th day, the Golgi apparatus is found in perinuclear and peripheral cytoplasmic positions. The formation of granular endoplasmic reticulum occurs in the vicinity of the perinuclear Golgi apparatus. The differentiation of cell and nuclear forms requires approximately 20 days. The morphological changes of differentiation are discussed in relation to the participation of the Golgi apparatus in the differentiation of dendrites and in the formation of the granular endoplasmic reticulum.     

ISOLATION OF GERM CELL GOLGI APPARATUS FROM SEMINIFEROUS TUBULES OF RAT TESTES

Intact Golgi apparatus have been isolated with good purity from rat testis by a simplified sucrose gradient technique. The procedure is inherently selective in that most of the Golgi apparatus in the isolate are from germ cells in late spermatocyte or early spermatid development. No Sertoli cell Golgi apparatus are present in the fraction. Biochemical analyses showed that the enzyme N-acetylglucosamine galactosyltransferase is enhanced in the band containing Golgi apparatus. Because they are easy to isolate, and because they are involved with the formation of a specific internal cellular component (the acrosome), these Golgi apparatus will be useful objects for comparison with other kinds of Golgi apparatus and for other studies leading to a better understanding of the basic functioning of Golgi apparatus.     

Functional morphology of the head of the anabantoid teleost fish Helostoma temmincki

Osteology, myology and motion analysis of the head of the anabantoid fish Helostoma temmincki, a specialized filter feeder, has revealed six functional units: neurocranium, suspensory apparatus, opercular apparatus, hyoid apparatus, branchial apparatus and pectoral girdle. Interactions between the functional units take place through four couplings involved in opening and protruding the jaws. The first coupling is activated in the beginning of the opening cycle by the levator operculi muscle through the opercular apparatus, interoperculomandibular ligament and mandible. The second is activated during feeding by contraction of the sternohyoideus through the hyoid apparatus, interopercular, interoperculomandibular ligament and mandible. The third coupling is active during feeding and “kissing” by contraction of epaxial muscles through mediation of the neurocranium to the jaw apparatus. The fourth coupling is the only one active during air intake and involves contraction of the levator arcus palatini which abducts and rotates the suspensory apparatus forwards, causing the mandible to drop. The retention of isolated ancestral characters during mosaic evolution are explained in terms of the maintenance of couplings which represent functional associations of seemingly remote structures. When natural selection acts on one component of a functional unit or coupling, it essentially acts on all associated elements simultaneously causing character complexes to evolve in common evolutionary trends. It is feasible that functional analysis can separate primary from secondary evolutionary trends.     

Src regulates Golgi structure and KDEL receptor-dependent retrograde transport to the endoplasmic reticulum

The tyrosine kinase Src is present on the Golgi membranes. Its role, however, in the overall function and organization of the Golgi apparatus is unclear. We have found that in a cell line called SYF, which lacks the three ubiquitous Src-like kinases (Src, Yes, and Fyn), the organization of the Golgi apparatus is perturbed. The Golgi apparatus is composed of collapsed stacks and bloated cisternae in these cells. Expression of an activated form of Src relocated the KDEL receptor (KDEL-R) from the Golgi apparatus to the endoplasmic reticulum. Other Golgi-specific marker proteins were not affected under these conditions. Because of the specific effect of Src on the location of KDEL-R, we tested whether protein transport between ER and the Golgi apparatus involves Src. Transport of Pseudomonas exotoxin, which is transported to the ER by binding to the KDEL-R is accelerated by inhibition or genetic ablation of Src. Protein transport from ER to the Golgi apparatus however, is unaffected by Src deletion or inhibition. We propose that Src has an appreciable role in the organization of the Golgi apparatus, which may be linked to its involvement in protein transport from the Golgi apparatus to the endoplasmic reticulum.     

A low-cost automated device incorporating a hollow fiber filtration cartridge for large-scale production of ghosts from human erythrocytes

Readily available elements were used to build an automatic apparatus dedicated to the preparation of erythrocyte ghosts. The apparatus is designed around a low-cost re-usable hollow-fiber filtration cartridge (marketed for therapeutic plasmapheresis). The apparatus is controlled by a simple programmer (based on a diode matrix and low cost timers and liquid level sensors): once the apparatus is loaded with whole red blood cells, washing of cells, as well as hemolysis and washing of ghosts, is performed by the machine in about 4.5 h without any operator intervention. Automatic filter cleaning takes a further 110 min.     

A compact apparatus for vertical gel slab isoelectric focusing: resolution of rabbit anti-pneumococcal polysaccharide antibodies.

A compact apparatus for analytical and preparative vertical gel slab isoelectric focusing is described. The apparatus was extremely versatile, easy to use, and comparatively inexpensive. The apparatus was useful to compare the electrofocusing patterns of anti-pneumococcal polysaccharide antibodies from several different restricted and partially restricted rabbit responders.     

Functional changes in the anatomy of the pharyngeal jaw apparatus of Astatoreochromis alluaudi (Pisces, Cichlidae), and their effects on adjacent structures

Organisms are tightly packed with structures so architectonic interdependency of structures is an obvious aspect of integration. This aspect of functional morphology, however, has received remarkably little attention. The present paper presents an example of the spatial relations among several apparatuses in the head of the cichlid fish, Astatoreochromis alluaudi. It investigates the transformations of these apparatuses and their functions due to a change in the pharyngeal jaw apparatus resulting from a functional shift (insect eating to snail crushing or vice versa ). The volume of the pharyngeal jaw apparatus differs 55% between the insect eating- and the snail eating morph. The increase in volume of the pharyngeal jaw apparatus has an impressive number of spatial effects, both direct and indirect, on other structures. Reallocation of space within the pharyngeal jaw apparatus occurs. Total head volume increases 31% but a reallocation of space is still necessary as the increase of the opercular compartment where the pharyngeal jaw apparatus is situated compensates for only 59% of the volume increase of that. Not all spatial effects do impose constraints. Spatial constraints are avoided when one of the apparatuses can use a topographically different volume of space. The respiratory apparatus shows internal reallocations of space without loss of total volume. The same solution occurs for elements of the expansion apparatus and the buccal savity. The eyes are not influenced. Finally spatial effects can have positive repercussions. The muscles of the oral jaw apparatus increase in size. This may be an example of an epiphenomenon.     

蕨类植物精细胞运动器和细胞骨架的研究

介绍了近年来蕨类植物游动精子运动器和细胞骨架的研究进展.游动精子由配子体精子器中的非运动细胞发育形成,其分化过程包括了运动器官和细胞骨架的合成和组装.精子发生过程中形成的运动器的各部分结构包括鞭毛、基体、多层结构及附属结构;基体是细胞中新形成的结构,在不同类群的蕨类植物中分别由双中心粒、分支生毛体和生毛体产生.鞭毛、基体和多层结构中的微管带形成了游动精子三个独特的微管列阵,由于微管蛋白的后修饰作用这些微管列阵十分稳定;centrin是运动器中的重要成分,但功能尚不清楚,可能和细胞骨架及运动器的构建有关.     

Dynamic concentration challenges for biosensor characterization

A method and apparatus are described for characterization of the steady state and dynamic response of biosensors. The apparatus produces a steady stream of homogeneously mixed analyte whose concentration can be fixed at discrete values or varied continously. The device is ideally suited for continously operated biosensors, but is also effective for biosensors that operate in discrete sampling modes. The system permits simultaneous testing of several sensors and determination of the accuracy, precision and repeatability of sensor response. The characteristics of this testing apparatus were validated with ferrocyanide and glucose as indicators. As an example of use of the apparatus, concentration ramps were created and used to complement conventional step changes for characterizing an implantable glucose sensor. The ramp rate can be adjusted easily by scaling the apparatus to simulate the rate of concentration change anticipated during actual monitoring situations.     

Observations on the Golgi apparatus in living plasma cells and in erythroblasts with dark ground illumination

The living Golgi apparatus has been studied in very thin preparations with dark ground illumination. Two kinds of intimately mingled droplets are found in a structureless ground substance. The movements performed by the two droplet systems are described in detail. The Golgi apparatus is capable of changing shape and is mobile. An exchange of droplets was observable between the Golgi apparatus and the surrounding cytoplasm.     

Cleavage furrow establishment by the moving mitotic apparatus

The midpoint of the mitotic apparatus is fixed in the future division plane long before the division mechanism develops, and this static relationship has been considered essential in speculations concerning division mechanism establishment. The purpose of the present investigation was to determine whether prevention of the static relationship affects the establishment process. Sand dollar eggs were reshaped into cylinders by confinement in an elastic capillary tube and, beginning about 20 min before cleavage, the mitotic apparatus was kept in reciprocal motion by alternately compressing the poles. When the movement was continuous and the excursions were 25, 50 or 75 μm, furrow activity developed near the midpoint of the region underlain by the mitotic apparatus. The acuteness of the furrow decreased as the distance the mitotic apparatus was moved increased. When the movement was made discontinuous by allowing the mitotic apparatus to pause at the end of each excursion, the results depended upon the duration of the pause. Pauses 30 s long resulted in a single furrow formed in the midpoint of the entire region underlain by the mitotic apparatus. When the pauses were 45s long, furrowing activity developed in both regions where the mitotic apparatus was allowed to pause. The results indicated that the normal static relation between the mitotic apparatus midpoint and the division plane is unnecessary for division mechanism establishment. They also demonstrate that a restricted region of contractile activity can be established in the cortex despite experimentally induced spreading and dilution of mitotic apparatus effect.     

Specific organization of Golgi apparatus in plant cells

Microtubules, actin filaments, and Golgi apparatus are connected both directly and indirectly, but it is manifested differently depending on the cell organization and specialization, and these connections are considered in many original studies and reviews. In this review we would like to discuss what underlies differences in the structural organization of the Golgi apparatus in animal and plant cells: specific features of the microtubule cytoskeleton organization, the use of different cytoskeleton components for Golgi apparatus movement and maintenance of its integrity, or specific features of synthetic and secretory processes. We suppose that a dispersed state of the Golgi apparatus in higher plant cells cannot be explained only by specific features of the microtubule system organization and by the absence of centrosome as an active center of their organization because the Golgi apparatus is organized similarly in the cells of other organisms that possess the centrosome and centrosomal microtubules. One of the key factors determining the Golgi apparatus state in plant cells is the functional uniformity or functional specialization of stacks. The functional specialization does not suggest the joining of the stacks to form a ribbon; therefore, the disperse state of the Golgi apparatus needs to be supported, but it also can exist “by default”. We believe that the dispersed state of the Golgi apparatus in plants is supported, on one hand, by dynamic connections of the Golgi apparatus stacks with the actin filament system and, on the other hand, with the endoplasmic reticulum exit sites distributed throughout the endoplasmic reticulum.     

一种新型半导体激光治疗机研制

采用二合一光纤系统,高效率光纤耦合装置,AlGaInp半导体激光器, 方便适用的光纤定位装置以及稳定可靠的电源控制系统,研制了一种新型650nm半导体激光治疗仪,其光纤端输出达109mW。该仪器已在生物医学领域成功应用。     

96-Well apparatus and method for use in enzyme-linked immunofiltration assay (ELIFA)

Summary An apparatus and method are described for use in enzyme-linked immunofiltration assay (ELIFA). The method of filtering ligands, blockers, wash buffers, and substrates decreases a typical 30 min to 2 hr incubation step with ELISA to 5 minutes. The apparatus differs from a dot-blot apparatus in that soluble coloured product is transferred directly into a receiving 96-well microtiter plate for quantifying with an automatic plate reader. Model systems of streptavidin/biotin and antibody/antigen form detectable complexes in the apparatus.     

The Golgi apparatus remains associated with microtubule organizing centers during myogenesis

In vitro myogenesis involves a dramatic reorganization of the microtubular network, characterized principally by the relocalization of microtubule nucleating sites at the surface of the nuclei in myotubes, in marked contrast with the classical pericentriolar localization observed in myoblasts (Tassin, A. M., B. Maro, and M. Bornens, 1985, J. Cell Biol., 100:35-46). Since a spatial relationship between the Golgi apparatus and the centrosome is observed in most animal cells, we have decided to follow the fate of the Golgi apparatus during myogenesis by an immunocytochemical approach, using wheat germ agglutinin and an affinity-purified anti-galactosyltransferase. We show that Golgi apparatus in myotubes displays a perinuclear distribution which is strikingly different from the polarized juxtanuclear organization observed in myoblasts. As a result, the Golgi apparatus in myotubes is situated close to the microtubule organizing center (MTOC), the cis-side being situated at a fixed distance from the nuclear envelope, a situation which suggests the existence of a structural association between the Golgi apparatus and the nuclear periphery. This is supported by experiments of microtubule depolymerization by nocodazole, in which a minimal effect was observed on Golgi apparatus localization in myotubes in contrast with the dramatic scattering observed in myoblasts. In both cell types, electron microscopy reveals that microtubule disruption generates individual dictyosomes; this suggests that the connecting structures between dictyosomes are principally affected. This structural dependency of the Golgi apparatus upon microtubules is not apparently accompanied by a reverse dependency of MTOC structure or function upon Golgi apparatus activity. Golgi apparatus modification by monensin, as effective in myotubes as in myoblasts, is without apparent effect on MTOC localization or activity and on microtubule stability. The main result of our study is to show that in a cell type where the MTOC is dissociated from centrioles and where antero-posterior polarity has disappeared, the association between the Golgi apparatus and the MTOC is maintained. The significance of such a tight association is discussed.     

Ultrastructural alteration of cytolytic T lymphocytes following their interaction with target cells. I. Hypertrophy and change of orientation of the Golgi apparatus.

The ultrastructure of cytolytic T lymphocytes adhered to the surface of target cells was investigated at different periods after start of interaction. Fifteen-minute incubation led to increase of number of Golgi apparatus cisternae and vacuoles. After 30 min incubation Golgi apparatus become oriented to the contact area. If several lymphocytes adhered to one target cell the Golgi apparatus of each of them was oriented toward the contact area. If one lymphocyte adhered simultaneously to two target cells its Golgi apparatus was oriented toward both target cells. Giant Golgi apparatus vacuoles were formed 30 to 60 min later and then moved to plasma membrane of lymphocyte and then the content of those vacuoles moved to the intercellular space between a cytolytic T lymphocyte and a target cell. The period required for the hypertrophy and change of orientation of Golgi apparatus is supposed to represent the “mobilization” step of a medium-sized and small killer lymphocyte.