Larval Development of Cucumaria Elongata (Echinodermata: Holothuroidea)

Author(s):  
Fu-Shiang Chia ◽  
J. B. Buchanan

Cucutnaria elongata Düben & Koren, collected from the Northumberland coast in January 1968, was induced to spawn in the laboratory by raising the temperature. At spawning, the animal waved its pharyngeal bulb back and forth while a thin string of gametes was discharged. The freshly spawned eggs, surrounded by a jelly coat, were at the end of the second meiotic division and the size varies from 250 to 350 μ in diameter. Oocytes in the ovarian tubules can be grouped into three stages according to their size and other morphological features. Insemination of artificially recovered eggs resulted in only a very low frequency of abnormal development up to 8-cell stage. However, over 90% fertilization was achieved by insemination of the naturally shed eggs and the development was normal. The cleavage was holoblastic and radial type. The larva hatched at post blastula stage and swam freely with its animal pole upward. A doliolaria stage with four ciliary bands and pentactula stage with five primary tentacles subsequently developed. The planktonic life at temperature 9–11 °C ended on the ninth day after fertilization. Pentactula, up to 4 months old, live underneath the mud where they are deposit feeders. The primary tentacles bearing mucus glands in the form of papillae are important in both locomotion and feeding. An outline of chronology from fertilization up to 5 months old is presented.

1985 ◽  
Vol 63 (8) ◽  
pp. 1989-1990
Author(s):  
Panagiotis A. Tsonis

The effects of a carcinogen, N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), on newt embryonic development were studied. When embryos are treated with MNNG before the blastula stage, abnormal development occurs. The most prominent effect is that the hinder region of the egg–embryo (vegetal pole) does not participate in the development; thus, only one hemisphere (animal pole) of the egg develops. This phenomenon is evident at the gastrula stage and becomes even more apparent during the neurula stage.


Development ◽  
1989 ◽  
Vol 107 (Supplement) ◽  
pp. 141-148 ◽  
Author(s):  
J. M. W. Slack ◽  
B. G. Darlington ◽  
L. L. Gillespie ◽  
S. F. Godsave ◽  
H. V. Isaacs ◽  
...  

In early amphibian development, the mesoderm is formed around the equator of the blastula in response to an inductive signal from the endoderm. A screen of candidate substances showed that a small group of heparin-binding growth factors (HBGFs) were active as mesoderm-inducing agents in vitro. The factors aFGF, bFGF, kFGF and ECDGF all show similar potency and can produce inductions at concentrations above about 100 pM. The product of the murine int-2 gene is also active, but with a lower specific activity. Above the induction threshold there is a progressive increase of muscle formation with dose. Single blastula ectoderm cells can be induced and will differentiate in a defined medium to form mesodermal tissues. All inner blastula cells are competent to respond to the factors but outer cells, bearing oocyte-derived membrane, are not. Inducing activity can be extracted from Xenopus blastulae and binds to heparin like the previously described HBGFs. Antibody neutralization and Western blotting experiments identify this activity as bFGF. The amounts present are small but would be sufficient to evoke inductions in vivo. It is not yet known whether the bFGF is localized to the endoderm, although it is known that inducing activity secreted by endodermal cells can be neutralized by heparin. The competence of ectoderm to respond to HBGFs rises from about the 128-cell stage and falls again by the onset of gastrulation. This change is paralleled by a rise and fall of binding of 125I-aFGF. Chemical cross-linking reveals that this binding is attributable to a receptor of relative molecular mass about 130 × 103. The receptor is present both in the marginal zone, which responds to the signal in vivo, and in the animal pole region, which is not induced in vivo but which will respond to HBGFs in vitro. In the embryo, the induction in the vicinity of the dorsal meridian is much more potent than that around the remainder of the marginal zone circumference. Dorsal inductions contain notochord and will dorsalize ventral mesoderm with which they are later placed in contact. This effect might be due to a local high bFGF concentration or, more likely, to the secretion in the dorsal region of an additional, synergistic factor. It is known that TGF-β-1 and -2 can greatly increase the effect of low doses of bFGF, although it has not yet been demonstrated that they are present in the embryo. Lithium salts have a dorsalizing effect on whole embryos or on explants from the ventral marginal zone, and also show potent synergism when applied together with HBGFs.


Development ◽  
1994 ◽  
Vol 120 (9) ◽  
pp. 2619-2628 ◽  
Author(s):  
R. Kuraishi ◽  
L. Osanai

Contribution of maternal cytoplasmic factors and cellular interaction to determination of archenteron in a starfish embryo was analyzed by (1) examining temporal and positional pattern of expression of an endoderm-specific enzyme, alkaline phosphatase, (2) deleting the vegetal polar fragment from an immature oocyte and (3) changing the orientation of a blastomere within an early stage embryo. The archenteron (and the differentiated digestive tract) of Asterina pectinifera was divided into three areas based on the time of start of alkaline phosphatase expression. At 27 hours after 1-methyladenine treatment, the whole archenteron except the anterior end started to express alkaline phosphatase. The anterior negative area differentiated into mesodermal tissues such as mesenchyme cells and anterior coelomic pouches (anterior mesodermal area). The alkaline-phosphatase-positive area 1 gave rise to the esophagus and the anterior end of the stomach. Alkaline-phosphatase-positive area 2, which was gradually added to the posterior end of the archenteron after 30 hours, became alkaline-phosphatase- positive and formed the middle-to-posterior part of the stomach and the intestine. When the vegetal oocyte fragment, the volume of which was more than 8% of that of the whole oocyte, was removed from the immature oocyte, archenteron formation was strongly suppressed. However, when the volume deleted was less than 6%, most of the larvae started archenteron formation before the intact controls reached the mesenchyme-migration stage (30 hours). Although cells in the alkaline-phosphatase-positive area 2 are added to the posterior end of the archenteron after 30 hours in normal development (R. Kuraishi and K. Osanai (1992) Biol. Bull. Mar. Biol. Lab., Woods Hole 183, 258–268), few larvae started gastrulation after 30 hours. Estimation of the movement of the oocyte cortex during the early development suggested that the area that inherits the cortex of the 7% area coincides with the combined area of anterior mesodermal area and alkaline-phosphatase-positive area 1. When one of the blastomeres was rotated 180° around the axis of apicobasal polarity at the 2-cell stage to make its vegetal pole face the animal pole of the other blastomere, two archentera formed at the separated vegetal poles. Intracellular injection of tracers showed that cells derived from the animal blastomere, which gives rise to the ectoderm in normal development, stayed in the outer layer until 30 hours; a proportion of them then entered the archenteron gradually. The involuted animal cells expressed alkaline phosphatase and were incorporated into the middle-to-posterior part of the stomach and the intestine. These results suggest that anterior mesodermal area and alkaline-phosphatase-positive area 1 are determined by cytoplasmic factor(s) that had already been localized in their presumptive areas. In contrast, alkaline-phosphatase-positive area 2 becomes the endoderm by homoiogenetic induction from the neighboring area on the vegetal side, namely alkaline-phosphatase-positive area 1.


Development ◽  
1997 ◽  
Vol 124 (13) ◽  
pp. 2553-2560 ◽  
Author(s):  
R. Maeda ◽  
A. Kobayashi ◽  
R. Sekine ◽  
J.J. Lin ◽  
H. Kung ◽  
...  

This study analyzes the expression and the function of Xenopus msx-1 (Xmsx-1) in embryos, in relation to the ventralizing activity of bone morphogenetic protein-4 (BMP-4). Expression of Xmsx-1 was increased in UV-treated ventralized embryos and decreased in LiCl-treated dorsalized embryos at the neurula stage (stage 14). Whole-mount in situ hybridization analysis showed that Xmsx-1 is expressed in marginal zone and animal pole areas, laterally and ventrally, but not dorsally, at mid-gastrula (stage 11) and late-gastrula (stage 13) stages. Injection of BMP-4 RNA, but not activin RNA, induced Xmsx-1 expression in the dorsal marginal zone at the early gastrula stage (stage 10+), and introduction of a dominant negative form of BMP-4 receptor RNA suppressed Xmsx-1 expression in animal cap and ventral marginal zone explants at stage 14. Thus, Xmsx-1 is a target gene specifically regulated by BMP-4 signaling. Embryos injected with Xmsx-1 RNA in dorsal blastomeres at the 4-cell stage exhibited a ventralized phenotype, with microcephaly and swollen abdomen. Histological observation and immunostaining revealed that these embryos had a large block of muscle tissue in the dorsal mesodermal area instead of notochord. On the basis of molecular marker analysis, however, the injection of Xmsx-1 RNA did not induce the expression of alpha-globin, nor reduce cardiac alpha-actin in dorsal marginal zone explants. Furthermore, a significant amount of alpha-actin was induced and alpha-globin was turned off in the ventral marginal zone explants injected with Xmsx-1. These results indicated that Xmsx-1 is a target gene of BMP-4 signaling, but possesses a distinct activity on dorsal-ventral patterning of mesodermal tissues.


Development ◽  
1973 ◽  
Vol 29 (3) ◽  
pp. 601-615
Author(s):  
M. H. L. Snow

Mouse embryos were grown in vitro from the 2-cell stage to blastocysts in the presence of [3H]thymidine. Methyl-T-thymidine and thymidine-6-T(n) were used and both forms found to be lethal at concentrations above 0·1 μCi/ml. Both forms of [3H]Tdr at concentrations between 0·01 and 0·1 μCi/ml caused a highly significant (P < 0·001) reduction in blastocyst cell number. The reduction in cell number, which was positively correlated with specific activity and tritium concentration, was associated with cell damage typical of radiation damage caused by tritium disintegration. Thymidine-6-T(n) also significantly reduced the number of 2-cell embryos forming blastocysts whereas methyl-T-Tdr did not. This difference in effect is assumed to be caused by contamination of one form of [3H]Tdr with a by-product of the tritiation process. A study of the cleavage stages showed that almost all the reduction in cell numbers could be accounted for by selective cell death occurring at the 16-cell stage. Cells which survive that stage cleave at a normal rate. The cells that are most susceptible to [3H]Tdr damage were found to normally contribute to the inner cell mass. The [3H]Tdr-resistant cells form the trophoblast. It is possible to grow blastocysts in [3H]Tdr such that they contain no inner cell mass but are composed entirely of trophoblast. Comparatively short (12 h) incubation with [3H]Tdr at any stage prior to the 16-cell stage will cause this damage. Possible reasons for this differential effect are discussed, and also compared with damage caused by X-irradiation.


2019 ◽  
Vol 22 (8) ◽  
pp. 1855-1866 ◽  
Author(s):  
Gang Li ◽  
Zhi-Qiang Huang ◽  
Zhang-Hua Lian ◽  
Lei Hao

To analyze the behavior of the vibrator–ground coupling vibration, a model containing equivalent dynamic stiffness and equivalent dynamic damping to describe the interaction between the vibrator and the ground is established based on half-space theory. According to load cell test, this model shows a good agreement with the experimental data. Dynamic responses of the structure are analyzed on displacement, velocity, acceleration, and ground force. Results show that the stroke and pump displacement are main constraints that limit the bandwidth of vibrator toward low frequency, and the stroke of conventional vibrator is not long enough to achieve lower frequency. Analysis of velocity response indicates that with the increase of frequency, a larger mass results in a lower velocity under external force. The influence of the ground acting on the baseplate is limited, and the acceleration of the baseplate is determined by its own mass beyond 80 Hz. Analysis of ground force shows that the response of the structure can be divided into three stages. The reaction mass, the baseplate, and the ground play different roles in dominating the ground force at different frequency bands.


Zygote ◽  
1999 ◽  
Vol 8 (S1) ◽  
pp. S42-S43 ◽  
Author(s):  
Tetsuya Kominami

Sea urchin pluteus larvae contain dozens of pigment cells in their ectoderm. These pigment cells are the descendants of the veg2 blastomeres of the 60-cell stage embryo. According to the fate map made by Ruffins and Ettensohn, the prospective pigment cells occupy the central region of the vegetal plate. Most of these prospective pigment cells exclusively give rise to pigment cells. Therefore, specification of the pigment cell lineage should occur at some point between the 60-cell and mesenchyme blastula stage. However, the detailed process of the specification of the pigment lineage is unknown.When are pigment cells specified? Are cell interactions necessary for the specification? Do founder cells exist? To answer these questions, I treated embryos with Ca2+-free seawater during the cleavage stage and examined the number of pigment cells observed in pluteus larvae. Treatment at 5.5–8.5 h and especially 7.5–10.5 h postfertilisation markedly reduced the number of pigment cells. The decrease was statistically significant. On the other hand, the treatment at 3.5–6.5 h or 9.5–12.5 h never reduced the number of pigment cells. By examining the frequency of the appearance of embryos whose numbers of pigment cells were less than 20, it was also found that the numbers of pigment cells were frequently in multiples of 4. Embryos having 4, 8, 12, 16 and 20 pigment cells were more frequently observed. Statistics indicated that the frequency of appearance was not random. These results indicated that cell contacts are necessary for the specification of pigment cells and that the specification occurs from 7 to 10 h postfertilisation. The results also suggest that the founder cells, if they exist, divide twice before they differentiate into pigment cells.


Zygote ◽  
1999 ◽  
Vol 8 (S1) ◽  
pp. S41-S41 ◽  
Author(s):  
David R. McClay

It has long been recognized that micromeres have special significance in early specification events in the sea urchin embryo. Micromeres have the ability to induce a secondary axis if transferred to the animal pole at the 16-cell stage of sea urchin embryos (Hörstadius, 1939). Without micromeres an isolated animal hemisphere develops into an ectodermal ball called a dauer blastula. Addition of micromeres to an animal half rescues a normal pluteus larva, including endoderm (Hörstadius, 1939). Despite these well-known experiments, however, neither the molecular basis of that induction nor the endogenous inductive role of micromeres in development was known. In recent experiments we learned that if one eliminates micromeres from the vegetal pole at the 16-cell stage the resulting embryo makes no secondary mesenchyme. Earlier it had been found that β-catenin is crucial for specification events that lead to mesoderm and endoderm (Wikra-manayake et al., 1998; Emily-Fenouil et al., 1998; Logan et al., 1999). We noticed that at the 16-cell stage β-catenin enters the nuclei of micromeres, then enters the nuclei of macromeres at the 32-cell stage (Logan et al., 1999). Since nuclear entry of β-catenin is known to be important for its signalling function in the Wnt pathway, we asked whether β-catenin functions in the micromere induction pathway.


2007 ◽  
Vol 19 (1) ◽  
pp. 240
Author(s):  
O. D. Owiny ◽  
D. M. Barry ◽  
M. Agaba ◽  
R. A. Godke

Interspecies hybridization of bovids occurs between domestic cattle and at least 3 other species: the American bison (Bison bison), yak (Bos grunniens), and banteng (Bos banteng). Birth of a cattle � buffalo hybrid was reported in Russia, but the report was never authenticated. Such hybrids could be important in improving livestock production and managing diseases that impede production in tropical Africa. We investigated hybridization between cattle and their closest African wild relative, the African buffalo (Syncerus caffer caffer). In an attempt to produce pre-implantation cattle � buffalo hybrid embryos in vitro, matured bovine oocytes were subjected to a standard IVF procedure with either homologous (n = 1166 oocytes) or heterologous (n = 1202 oocytes) buffalo epididymal sperm. After IVF, 67.2% of the oocytes inseminated with homologous sperm cleaved. In contrast, insemination with buffalo sperm resulted in a 4.6% cleavage rate. Cleavage was also slower in hybrids than in cattle embryos. Up to 52.2% of cleaved homologous embryos progressed to the morula stage compared with 12.7% for hybrids. No hybrid embryos developed beyond the 16-cell stage, whereas 40.1% of the cleaved bovine embryos developed to the blastocyst stage. Developmental anomalies such as polyspermy, uneven cleavage, vacuolization, and absence of nuclei in some blastomeres were common in the hybrid embryos. We conclude that interspecies fertilization of cattle oocytes with African buffalo sperm occurs in vitro and that the barrier to hybridization is in the early stages of embryonic development. Also, chromosomal disparity is the likely cause of fertilization abnormalities, abnormal development, and subsequent arrest, impairing the formation of pre-implantation hybrid embryos. Investigation into developmental abnormalities, including reciprocal hybridization and genetic studies of the hybrid embryos, are recommended.


Development ◽  
1989 ◽  
Vol 106 (4) ◽  
pp. 641-647 ◽  
Author(s):  
R.A. Cameron ◽  
S.E. Fraser ◽  
R.J. Britten ◽  
E.H. Davidson

Several lines of evidence suggest that the oral-aboral axis in Strongylocentrotus purpuratus embryos is specified at or before the 8-cell stage. Were the oral-aboral axis specified independently of the first cleavage plane, then a random association of this plane with the blastomeres of the four embryo quadrants in the oral-aboral plane (viz. oral, aboral, right and left) would be expected. Lineage tracer dye injection into one blastomere at the 2-cell stage and observation of the resultant labeling patterns demonstrates instead a strongly nonrandom association. In at least ninety percent of cases, the progeny of the aboral blastomeres are associated with those of the left lateral blastomeres and the progeny of the oral blastomeres with the right lateral ones, respectively. Thus, ninety percent of the time the oral pole of the future oral-aboral axis lies 45 degrees clockwise from the first cleavage plane as viewed from the animal pole. The nonrandom association of blastomeres after labeling of the 2-cell stage implies that there is a mechanistic relation between axis specification and the positioning of the first cleavage plane.


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