怀头鲇成熟卵膜表面扫描电镜观察

The surface ultrastructure of mature egg chorion of the tamed northern sheatfish Silurus soldatovi in Hei- longjiang river was investigated by scanning electron microscopy. The outermost layer was composed of a thin lamella of a jelly coat. The surface of ielly coat was characterized by overlapping net-like fibrillar lamellae, many clefts with bridges and pores. The pore canals were 0.25 μm in diameter. The pore density was about 0.7 - 1.0/μm^-2. The pores on the surface of jelly coart were distributed uniformly. Mature eggs of sheatfish possessed a funnel-shaped micropyle in the animal polar region. The micropylar region was about 0.04 mm^2 area in diameter with 3 - 8 ridges in radial arrangement. Fertilized eggs in which jelly coats were removed mechanically presented no adhesiveness and exhibited inlays of irregular granules on their surface [Acta Zoologica Sinica 51 （5）： 940-946, 2005].     

Observation of nuclei reassembled from demembranated Xenopus sperm nuclei and analysis of their lamina components

A cell-free preparation obtained from extracts of activated Xenopus laevis eggs induced chromatin decondensation and nuclear formation from demembranated Xenopus sperm nuclei.Electron microscopy revealed that the reassembled nucleus had a double-layered nuclear memblane,nuclear pore complexes,and decondensed chromatin etc.Indirect immunofluorescence analysis demonstrated the presence of lamina in newly assembled nuclei.Western-blotting results showed that lamin LII was present in egg extracts and in lamina of the reassembled nuclei which were previously reported to contain only egg derived lamin LIII.     

Ultrastructure of the Mature Egg and Fertilization in the Fern Ceratopteris thalictroides

The ultrastructure of the mature egg and fertilization in the fern Ceratopteris thafictroides （L.） Brongn. were observed by transmission electron microscopy. The results revealed that the mature egg possesses an obvious egg membrane at the periphery of the egg. Furthermore, a fertilization pore was identified in the upper egg membrane of the mature egg. The structure of the pore is described for the first time. The fertilization experiment indicated that spermatozoids crowd into the cavity above the egg through the neck canal of the archegonium; however, only one of these can penetrate into the egg through the fertilization pore. Immediately on penetration of the spermatozoid, the egg begins to shrink. The volume of the fertilized egg decreases to almost one-half that of the unfertilized egg. As a result, the protoplasm of the fertilized egg becomes dense and opaque, which may lead to a situation where the organelles of both the egg and the fertilizing spermatozoid become indistinguishable. Simultaneously, abundant vesicles containing concentric membranes or opaque materials appear near the fertilization pore in the cytoplasm of the fertilized egg. These vesicles are considered to act as a barrier that prevents polyspermy. The present study provides a new insight into the ultrastructure of the mature egg and the cytological mechanism of fertilization in ferns.     

Electrofusion between Blastula Cells and Unfertilized Egg in Loach (Paramisgurnus dabryanus)

Electrofusion between blastula cells and unfertilized eggs in loach were investigated usingdielectrophoretic field where, under alternating sinusoidal electric field, blastula cells formed beads-like chain in close contact with the unfertilized egg and cell fusion occurred between eggs and thecells in tight contact with them. The nuclei ofblastula cells were brought into the cytoplasm of therecipient eggs, where they promoted the development of the fused eggs just like the zygote nuclei.But the development of the fused eggs was different from that of zygotes. Several nuclei might enterone and the same egg simultaneously and all of them could undergo division, resulting in severalblastomere after the first cleavage of the recipient egg. Before blastula stage, the embryo developingfrom the fused egg showcd irregular shape, but it was soon regulated and developed to a normalblastula which often continued its development into a normal individual. Cell/egg electrofusion cameto its highest fosion rate (80%) 8nd hatching rate (20%), with cell density at 1×10~3 cells/ml, Ca~(++)concentration at 10 mM, mannitol at 0.2 M and when the blastula cells were digested with 100μg/ml pronase E for 6-10 min at 20℃. The mechanism underlying development of electrofused eggsis discussed. As the result indicates, electrofusion might prove to be a promising biotechnology justas nuclear transplantation.     

Ecdysteroids in the adults and eggs of Opogona sacchari (Bojer), an invasive alien pest

In order to understand the composition and quantitative variation of ecdysteriods in the adults and eggs of Opogona sacchari (Bojer), an invasive alien pest, we analyzed the ecdysteroid composition and titers in the adult and egg of this pest. On day 4 after eclosion, the titer of ecdysteriods in the male adult was 0.080 ng/adult, much lower than 5.978 ng/adult in the female adult. During the development of ovaries, the titer of ecdysteriods was low on the first two days, and high in the late period, with the peak (10.48 ng/ovary) appearing on day 3. During the development of eggs, the titer of ecdysteriods was about 0.010 ng/egg from day 1 to day 3, and then decreased to 0.006 ng/egg on day 4. In both adults and eggs, three main components of ecdysteriods were found by identification of HPLC/RIA. They were 20-hydroxyecdysone, 26-hydroxyecdysone, and an unidentified component.     

The giant panda (Ailuropoda melanoleuca) somatic nucleus can dedifferentiate in rabbit ooplasm and support early development of the reconstructed egg

The giant panda skeletal muscle cells, uterus epithelial cells and mammary gland cells from an adult individual were cultured and used as nucleus donor for the construction of interspecies embryos by transferring them into enucleated rabbit eggs. All the three kinds of somatic cells were able to reprogram in rabbit ooplasm and support early embryo development, of which mammary gland cells were proven to be the best, followed by uterus epithelial cells and skeletal muscle cells. The experiments showed that direct injection of mammary gland cell into enucleated rabbit ooplasm, combined with in vivo development in ligated rabbit oviduct, achieved higher blastoeyst development than in vitro culture after the somatic cell was injected into the perivitelline space and fused with the enucleated egg by electrical stimulation. The chromosome analysis demonstrated that the genetic materials in reconstructed blastocyst cells were the same as that in panda somatic cells. In addition, giant panda mitochondrial DNA (     

Binding of epithelial cells to lectin-coated surfaces

Summary Epithelial cells may relate to their basement membrane substrates via lectin-like interactions. In a model system for study of this type of interaction, lectin-coated bacteriological plastic petri dishes were presented as substrates for epithelial cell adhesion. Of 21 lectins tested by mixed agglutination against two epithelial cell types, Madin-Darby canine kidney (MDCK), and human embryonic kidney cells (HEK), nine gave less than 5% rosettes and 12 gave 5 to 50% rosettes. Wheat germ agglutinin (WGA) andGeodia cydonium lectin gave the highest percentage of rosettes. Wheat germ agglutinin was readily adsorbed to plastic surfaces and maintained specificity in binding interactions. Both MDCK and HEK cells attached as well to WGA coated petri dishes as to conventional tissue culture dishes. Furthermore, both spread over the lectin-coated surfaces. The MDCK cells grew to confluence and could be subcultured and maintained indefinitely on such surfaces, although WGA in solution was toxic to the cells in concentrations as low as 0.1 to 1.0 μg/ml. Cell attachment to WGA coated dishes was blocked by cycloheximide only if the cells had been preincubated with the inhibitor for several hours. Cell attachment was not inhibited by pretreatment of cells with neuraminidase. Precoating cells with WGA blocked binding to both WGA-coated surfaces and untreated tissue culture dishes. Cells attached to WGA-coated dishes could not be readily dislodged by trypsin-EDTA for the first 2 h after subculture. By 4 h, attachment was again trypsin sensitive, suggesting that the cells synthesized a trypsin-sensitive material that was laid down between the cell surface and the WGA-coated dish. Regeneration of trypsin sensitivity was not blocked by cycloheximide. This work was supported by Research Grant AG01986 from the National Institutes of Health, Bethesda, Maryland.     

中国南方海域鲎的种类和分布

The northern sea area of the South China Sea is situated between 107.5°～122.5°East longitude and 17°～25° North latitude with high temperature and a large quantity of rainfall which belongs to the oceanic climate of the torrid zone and subtropical zone. From September, 1994 to June, 1998,during the research on the species of Horseshoe crab within the scope between the south from North gulf and the north to Xiamen, choosing Xiamen, Shantou, Wuchuan, Donghaidao and Qishui as fixed positions and irregularly going to Xuwen and Suixi in Guangdong province and Fangcheng in Guangxi province, we found that there was only Tachypleus tridentatus in the northern sea area of the South China Sea and the sea area from Wuchuan to Xiamen, and found that there were two species of horseshoe crab which were Tachypleus tridentatus and Carcinoscorpius rotundicauda in the southern area to East Sea Island, referring to East Sea Island, Qishui, Xuwen, Suixi and Fangcheng, most of which were Tachypleus tridentatus, and found a great number of larvae of Tachypleus tridentatus including zooids with different sizes from 7cm bodily length to mature. In Minan we found not only big colonies of adult Carcinoscorpius rotundicauda , but also a great number of larvae of Carcinoscorpius rotundicauda with different body size from 7cm body length to maturity. All these show that the northern sea area of the South China Sea is the area where both Tachypleus tridentatus and Carcinoscorpius rotundicauda distribute and reproduce. On January 1st, 1998 at the town of Qishui in Leizhou, we found an adult male Horseshoe crab and majority of its eggs was immature, while the eggs with above 4mm diameters were only 0.87 percent but the eggs with below 1.0mm diameter was 58 percent(1 160 pills). It shows that Qishui is a place where Tachyplers tridentatus survive in the winter.     

Seasonal movement of the American horseshoe crab Limulus polyphemus in a semi-enclosed bay on Cape Cod,Massachusetts (USA) as determined by acoustic telemetry

<正> American horseshoe crabs Limulus polyphemus were tracked using acoustic telemetry and traditional tagging in asemi-enclosed bay on Cape Cod (Pleasant Bay),Massachusetts,USA,to determine seasonal movement patterns.Fifty-five activelyspawning females were fitted with transmitters in 2008 and 2009 and were tracked using acoustic telemetry from May 2008through July 2010.Fifteen crabs with transmitters also had archive depth-temperature tags attached.In addition,over 2000spawning crabs (males and females) were tagged with US Fish and Wildlife (USFWS) button tags over the same period.Ninety-one percent of the crabs with transmitters were detected during this study.In the spring,crabs were primarily located inthe northern section of the bay near spawning beaches,whereas in the fall crabs moved towards the deeper portions of the bay,and some may have overwintered in the bay.There was evidence that a majority (58%-71%) of the females with transmittersspawned in two sequential seasons.One archive tag was recovered resulting in a year-long continuous record of depth and temperaturedata that,when integrated with telemetry data,indicated that the crab overwintered in the bay.The live recapture rate ofcrabs with USFWS button tags was 11%,with all re-sighted crabs except one observed inside Pleasant Bay.Eighty-three percentof recaptures were found within 2.5km of the tagging location,and 51% were observed at the same beach where they were tagged.This study provides further evidence that horseshoe crabs in Pleasant Bay may be philopatric to this embayment     

Recognition of nestmate eggs in the ant Formicafusca is based on queen derived cues

Inclusive fitness benefits depend on recognizing the right individuals to interact with. Social insect nests protect themselves from non-kin intruders through nestmate recognition based on chemical cues. The recognition cues on adult individu- als are from a mixture of genetic and environmental sources, but the ontogeny and use of recognition cues on eggs has not been previously assessed. We studied recognition by workers of eggs that were either nestmates or non-nestmates, and the ontogeny of recognition cues on eggs in the ant Formica fusca, a species with precise egg recognition abilities. Workers were able to dis- criminate among freshly laid eggs with no nest derived cues on them, and the egg surface chemicals varied among nests in these eggs, suggesting that queen derived cues are used in nestmate recognition. The results are discussed in the light of their implica- tions on deceptive social parasite strategies and within colony conflicts     

Characteristics of the membranes of the pellicle of the ciliatePseudomicrothorax dubius

Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles.     

The early development of the tail and the transformation of the shape of the nucleus of the spermatid of the domestic fowl,Gallus gallus

Summary The differentiation of the spermatid, especially in reference to the formation of the flagellum, and transformation of the shape of the nucleus was investigated in the domestic fowl.In the early stage of the spermatid, a prominent Golgi apparatus appears around the centrioles. The Golgi vesicles then surround the axial-filament complex which develops from the distal centriole. These vesicles fuse to form continuous membrane at the earliest stage of flagellar formation, and in the succeeding stage Golgi lamellae are attached to the plasma membrane of the developing flagellum. From these observations, it is assumed that Golgi apparatus may be a source of the membrane system of the flagellum.The microtubules distributed around the nucleus form the circular manchette. The anterior region of the nucleus with the manchette is cylindrical in shape and the posterior region without it remains irregular in shape. When the circular manchette has been completed, the whole nucleus acquires a slender cylindrical shape. The circular manchette then changes into the longitudinal manchette. The nuclei of spermatids without a longitudinal manchette are abnormal in shape. In view of these observations it is assumed that the nuclear shaping of the spermatid may be accomplished by circular manchette and the maintenance of shape of the elongated nucleus by longitudinal manchette.The authors wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices     

Characterization of the composition of the aqueous humor and the vitreous body of the eye of the frog Rana temporaria L

This study aimed to analyze the aqueous humor (AH) and the vitreous body (VB) of the eye of the adult frog Rana temporaria L. as a representative species of amphibians, which lead a semi-terrestrial life. The presence of collagen, albumin, uric acid and electron donors was shown in both media; however, there are slight differences in their concentrations. To determine collagen, a spectral-fluorescent probe, cyanine dye, was used. The presence of collagen in AH of the frog was found at the first time. The total content of electron donors (ascorbic and uric acids, tryptophan, and tyrosine) in VB and HA was roughly estimated at ~ 1.5 × 10^− 4 mol/L. Both VB and AH absorb light in similar UV regions. The total protein and albumin contents in AH were found to be somewhat higher than those in VB. The uric acid content was at an equally low level in both intraocular media. It is supposed that the similarity of VB and AH compositions shown in this work is due to some exchange between VB and AH contents in the course of accommodation. The role of intraocular fluids in physiological functions of the eye and in protecting the retina against UV light is discussed.     

Observations on the permeability of the choriocapillaris of the eye

Summary The choriocapillaris is a fenestrated capillary bed located posterior to the retinal pigment epithelium. It serves as the main source of supply to the photoreceptors, retinal pigment epithelium, and other cells of the outer retina. The permeability of these capillaries to intravenously injected ferritin (MW — approx. 480,000; mol. diam. 11 nm) was examined in the mouse, rabbit, and guinea pig, each of which is characterized by a different type of retinal vascularization. In all three species, the bulk of the ferritin remained in the capillary lumina, where it appeared to be blocked at the level of the diaphragmed fenestrae. Some ferritin was present in endothelial cell vacuoles. The results confirm previous work on the rat choriocapillaris and indicate that the barrier function of the choriocapillary endothelium is present even among species in which the retinal circulation differs significantly.Supported by NIH grant EY03418