Scanning electron microscopy of gastrulation in a sea urchin (Anthocidaris crassispina)

Development ◽  
1982 ◽  
Vol 67 (1) ◽  
pp. 27-35
Author(s):  
Shonan Amemiya ◽  
Koji Akasaka ◽  
Hiroshi Terayama

Gastrulation in Anthocidaris was investigated by observing the inside and the outside of embryos by scanning electron microscopy. Furrows which possibly rėflect changes in intercellular interactions were observed on the outer surface (hyaline layer side) of embryos twice in development: firstly at the time of primary mesenchyme cell formation, and secondly at the time of vegetal plate indentation. In the latter case, the cells within and surrounding the vegetal plate appeared to change their shapes differently; the former (within the plate) having broader surfaces on the blastocoel side whereas the latter (surrounding the plate) having broader surfaces on the hyaline layer side. This suggests that the first phase of indentation may be mediated by the autonomous change of cell shape and intercellular adhesiveness, accompanied by an autonomous cell movement in the vegetal pole region. Although some pseudopodial linkages were observed between secondary mesenchyme cells on the top of the invaginating archenteron and the animal pole in the mid-gastrula and later stage embryos, they were thinner and smaller in number as compared to those in the Pseudocentrotus embryos. The rate of invagination appeared rather constant throughout gastrulation in contrast to the accelerated invagination in other embryos with larger blastocoel cavities. Moreover, the number of columnar cells on the dissected surface of embryos remained unaltered. These findings suggest that the secondary mesenchyme cells may act as a linker between the archenteron tip and the animal pole, but they may not generate major motive forces for archenteron invagination at least in the Anthocidaris embryos.

Zygote ◽  
2009 ◽  
Vol 17 (2) ◽  
pp. 93-99 ◽  
Author(s):  
Luciana Nakaghi Ganeco ◽  
Irene Bastos Franceschini-Vicentini ◽  
Laura Satiko Okada Nakaghi

SummaryIn the present work, we analyzed the structure of oocytes and fertilized eggs of the piracanjuba fish (Brycon orbignyanus) under light and scanning electron microscopy. After inducing spawning, samples were collected at the moment of oocyte extrusion, when oocytes and semen were mixed (time 0), as well as at 10, 20 and 30 s after mixing, every minute up to 10 min, and then at 15 and 20 min. The oocytes are spherical, translucent and greenish with a mean diameter of 1.3 ± 0.11 mm. During the extrusion, cytoplasmic movement was observed in eggs towards the micropyle, characterizing the animal pole. At the moment of fertilization, the cortical cytoplasm showed a higher concentration of cortical alveoli at the animal pole than at the vegetal pole. The cortical alveoli breakdown promoted the elevation of the chorion with a consequent increase in egg diameter (1.95 ± 0.08 mm). The penetration of the spermatozoon promotes the formation of a fertilization cone of spherical external structure, which obstructs the opening of the micropyle. This structure acts as a main mechanism to avoid polyspermy, intercepting the access of supernumerary spermatozoa. Such studies about the reproductive biology of fish are important to species survival and conservation programmes.


2016 ◽  
Author(s):  
Chisato Kitazawa ◽  
Tsubasa Fujii ◽  
Yuji Egusa ◽  
Miéko Komatsu ◽  
Akira Yamanaka

ABSTRACTEmbryos of temnopleurid sea urchins exhibit species-specific morphologies. While Temnopleurus toreumaticus has a wrinkled blastula, others have a smooth blastula. Embryos of T. toreumaticus invaginate continuously at gastrulation, whereas in some others invagination is stepwise. We studied blastula and gastrula formation in four temnopleurids using light and scanning electron microscopy to clarify the mechanisms producing these differences. Unlike T. toreumaticus, blastomeres of mid-blastulae in T. reevesii, T. hardwickii and Mespilia globulus formed pseudopods. Before primary mesenchyme cells ingressed, embryos developed an area of orbicular cells in the vegetal plate. The cells surrounding the orbicular cells extended pseudopods toward the orbicular cell area in T. toreumaticus, T. reevesii and T. hardwickii. In T. toreumaticus, the extracellular matrix was well-developed and developed a hole-like structure that was not formed in others. Gastrulation of T. reevesii, T. hardwickii and M. globulus was stepwise, suggesting that differences of gastrulation are caused by all or some of factors: change of cell shape, rearrangement, pushing up and towing of cells. These species-specific morphologies may be caused by the shape and surface structure of blastomeres with cell-movement.Summary statement:Temonopleurid embryology


2021 ◽  
Author(s):  
Yixuan Wang ◽  
Si Zhang ◽  
Qian Sun ◽  
Fan’en Yuan ◽  
Linyao Zhao ◽  
...  

Abstract WAC is closely related to the occurrence and development of tumors. However, its role in human glioblastoma (GBM) and its potential regulatory mechanisms have not been investigated. This study demonstrated that WAC is downregulated in GBM, and its low expression predicts a poor prognosis. We investigated the effect of WAC on the proliferation of glioma cells through the CCK-8 assay, EdU incorporation and cell formation. The effects of WAC on apoptosis and autophagy in glioma were demonstrated by flow cytometry, TUNEL detection, immunofluorescence, q-PCR, WB and scanning electron microscopy. We found that overexpression of WAC inhibited proliferation of glioma cells, promoted apoptosis and induced autophagy. Therefore, WAC is likely to play a role as a new regulatory molecule in glioma.


Author(s):  
P.S. Porter ◽  
T. Aoyagi ◽  
R. Matta

Using standard techniques of scanning electron microscopy (SEM), over 1000 human hair defects have been studied. In several of the defects, the pathogenesis of the abnormality has been clarified using these techniques. It is the purpose of this paper to present several distinct morphologic abnormalities of hair and to discuss their pathogenesis as elucidated through techniques of scanning electron microscopy.


Author(s):  
P.J. Dailey

The structure of insect salivary glands has been extensively investigated during the past decade; however, none have attempted scanning electron microscopy (SEM) in ultrastructural examinations of these secretory organs. This study correlates fine structure by means of SEM cryofractography with that of thin-sectioned epoxy embedded material observed by means of transmission electron microscopy (TEM).Salivary glands of Gromphadorhina portentosa were excised and immediately submerged in cold (4°C) paraformaldehyde-glutaraldehyde fixative1 for 2 hr, washed and post-fixed in 1 per cent 0s04 in phosphosphate buffer (4°C for 2 hr). After ethanolic dehydration half of the samples were embedded in Epon 812 for TEM and half cryofractured and subsequently critical point dried for SEM. Dried specimens were mounted on aluminum stubs and coated with approximately 150 Å of gold in a cold sputtering apparatus.Figure 1 shows a cryofractured plane through a salivary acinus revealing topographical relief of secretory vesicles.


Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).


Author(s):  
Ronald H. Bradley ◽  
R. S. Berk ◽  
L. D. Hazlett

The nude mouse is a hairless mutant (homozygous for the mutation nude, nu/nu), which is born lacking a thymus and possesses a severe defect in cellular immunity. Spontaneous unilateral cataractous lesions were noted (during ocular examination using a stereomicroscope at 40X) in 14 of a series of 60 animals (20%). This transmission and scanning microscopic study characterizes the morphology of this cataract and contrasts these data with normal nude mouse lens.All animals were sacrificed by an ether overdose. Eyes were enucleated and immersed in a mixed fixative (1% osmium tetroxide and 6% glutaraldehyde in Sorenson's phosphate buffer pH 7.4 at 0-4°C) for 3 hours, dehydrated in graded ethanols and embedded in Epon-Araldite for transmission microscopy. Specimens for scanning electron microscopy were fixed similarly, dehydrated in graded ethanols, then to graded changes of Freon 113 and ethanol to 100% Freon 113 and critically point dried in a Bomar critical point dryer using Freon 13 as the transition fluid.


Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).


Author(s):  
J.N. Ramsey ◽  
D.P. Cameron ◽  
F.W. Schneider

As computer components become smaller the analytical methods used to examine them and the material handling techniques must become more sensitive, and more sophisticated. We have used microbulldozing and microchiseling in conjunction with scanning electron microscopy, replica electron microscopy, and microprobe analysis for studying actual and potential problems with developmental and pilot line devices. Foreign matter, corrosion, etc, in specific locations are mechanically loosened from their substrates and removed by “extraction replication,” and examined in the appropriate instrument. The mechanical loosening is done in a controlled manner by using a microhardness tester—we use the attachment designed for our Reichert metallograph. The working tool is a pyramid shaped diamond (a Knoop indenter) which can be pushed into the specimen with a controlled pressure and in a specific location.


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