The effect of u.v. irradiation of the vegetal pole of Xenopus laevis eggs on the presumptive primordial germ cells

Development ◽  
1975 ◽  
Vol 34 (1) ◽  
pp. 209-220
Author(s):  
Brigitta Züst ◽  
K. E. Dixon

The initial effect of u.v. irradiation of the vegetal pole was to inhibit cleavage in the vegetal hemisphere although karyokinesis was not substantially affected. In this way a syncytium formed in the vegetal hemisphere which broke down into individual cells some time between morula and late blastula. The movement of the germ plasm from the peripheral cortical regions into the interior of the egg was not appreciably delayed although aggregation of the germ plasm did not take place until the individual presumptive primordial germ cells were formed when the syncytium broke down. The method of segregation of the germ plasm and formation of the presumptive primordial germ cells was therefore very different in irradiated embryos from the normal orderly processes which depend on normal cleavage patterns. After neurula, the number of presumptive primordial germ cells declined rapidly and at stage 43/44, when the genital ridges in normal embryos contain primordial germ cells, the genital ridges in irradiated embryos were sterile. These results raise the question whether derangement of the segregation of the presumptive primordial germ cells is solely responsible for the later abnormalities in the cell lineage or whether u.v. irradiation affects the germ plasm and therefore indirectly the germ cells.

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 79-99
Author(s):  
J. H. Cleine ◽  
K. E. Dixon

Eggs of X. laevis were rotated (sperm entrance point downwards) either through 90° (1×90 embryos) or 180° in two 90° steps (2×90 embryos) at approximately 25–30 min postfertilization after cooling to 13°C. The embryos were kept in their off-axis orientation and cooled until the early gastrula stage. Rotation resulted in relocation of egg constituents with slight changes in the distribution of outer cortical and subcortical components and major changes in inner constituents where the heavy yolk and cytoplasm appeared to reorient as a single coherent unit to maintain their relative positions with respect to gravity. Development of rotated embryos was such that regions of the egg which normally give rise to posterior structures instead developed into anterior structures and vice versa. Germ plasm was displaced in the vegetal-dorsal-animal direction (the direction of rotation) and was segregated into dorsal micromeres and intermediate zone cells in 2×90 embryos and dorsal macromeres and intermediate zone cells in 1×90 embryos. In consequence, at the gastrula stage, cells containing germ plasm were situated closer to the dorsal lip of the blastopore after rotation — in 2×90 gastrulas around and generally above the dorsal lip. Hence, in rotated embryos, the cells containing germ plasm were invaginated earlier during gastrulation and therefore were carried further anteriorly in the endoderm to a mean position anterior to the midpoint of the endoderm. The number of cells containing germ plasm in rotated embryos was not significantly different from that in controls at all stages up to and including tail bud (stage 25). However at stages 46, 48 and 49 the number of primordial germ cells was reduced in 1×90 embryos in one experiment of three and in 2×90 embryos in all experiments. We tested the hypothesis that the decreased number of primordial germ cells in the genital ridges was due to the inability of cells to migrate to the genital ridges from their ectopic location in the endoderm. When anterior endoderm was grafted into posterior endodermal regions the number of primordial germ cells increased slightly or not at all suggesting that the anterior displacement of the cells containing germ plasm was not the only factor responsible for the decreased number of primordial germ cells in rotated embryos. Other possible explanations are discussed.


Development ◽  
1977 ◽  
Vol 39 (1) ◽  
pp. 221-233
Author(s):  
Masami Wakahara

Larvae of Rana chensinensis developed from fertilized eggs which had been subjected to ultraviolet (u.v.) irradiation on their vegetal hemisphere at a dose of 20000 ergs/mm2 within 60 min of fertilization contained no primordial germ cells (PGCs) when examined histologically at the stage when the operculum was complete (8 days after fertilization at 18 °C, stage 25 according to Shumway, 1940). The morphogenetic ability of vegetal pole cytoplasm from non-irradiated eggs to establish the PGCs was tested by injecting some fractions of this cytoplasm into the vegetal hemisphere of u.v.-irradiated eggs. Crude homogenate of the vegetal pole cytoplasm without large yolk platelets was able to restore the PGCs when injected into u.v.-irradiated eggs, but a similar fraction from animal half cytoplasm had no ability to form PGCs. The ‘PGC-forming activity’ demonstrated in the crude homogenate of the vegetal pole cytoplasm was not abolished by dialysis, lyophilization and heating to 90 °C for 10 min. When the homogenate was fractionated by differential centrifugation in 0·25 M sucrose, the ‘PGC-forming activity’ was recovered mainly in the precipitate of 15000g for 30 min. The precipitate of 7000 g for 10 min had also a little ‘activity’. The possibility was discussed that the ‘PGC-forming activity’ demonstrated in the vegetal pole cytoplasm was associated with the germinal granules in the germ plasm rather than the mitochondria.


Development ◽  
1977 ◽  
Vol 41 (1) ◽  
pp. 33-46
Author(s):  
Brigitta Züst ◽  
K. E. Dixon

Approximately 20–25 primordial germ cells leave the endoderm between stages 38–41 and localize in the dorsal root of the mesentery by stage 43/44. At this time all the cells contain large quantities of yolk which is gradually resorbed. The cells begin dividing between stages 48–52. The number and size of the germ cells were measured in tadpoles between stages 48–54 of development. The results indicate that in females the germ cells divide more often than in males. In both sexes the mitoses are grossly unequal, leading to the formation of a new generation of germ cells which are considerably smaller (one-tenth to one-fifth) than the size of the primordial germ cells at stage 48. The germ cells in male tadpoles at stage 54 are larger than in female tadpoles at the same stage. In tadpoles which developed from eggs irradiated in the vegetal hemisphere with u.v. light at the 2- to 4-cell-stage, primordial germ cells migrate into the genital ridges much later (stage 46–48) than in unirradiated embryos. They also differ morphologically from germ cells in control animals at this stage in that they are approximately one-tenth the size, lacking yolk in the cytoplasm and have a more highly lobed nucleus. Comparison of the results in unirradiated and irradiated animals suggests that the germ cell lineage is composed of a series of ordered, predictable events, and serious disruption of one of the events deranges later events.


Biology Open ◽  
2012 ◽  
Vol 2 (2) ◽  
pp. 191-199 ◽  
Author(s):  
T. Yamaguchi ◽  
A. Taguchi ◽  
K. Watanabe ◽  
H. Orii

Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 83-93
Author(s):  
J. H. Cleine

The genital ridges of Xenopus laevis tadpoles reared from eggs kept in an inverted position contain less than 40 % of the number of primordial germ cells (PGCs) of controls (Cleine & Dixon, 1985). It has been suggested that this reduction is caused by the germ cells' ectopic position in the anterior endoderm of larvae from inverted eggs, from where they may be unable to migrate into the genital ridges (Cleine & Dixon, 1985). This hypothesis is tested here by interchanging anterior and posterior endodermal grafts between pairs of inverted embryos at the early tailbud stage. Replacement of anterior by posterior endoderm has no effect but replacement of posterior by anterior endoderm increases the number of PGCs in the genital ridges and significantly reduces the proportion of sterile embryos. In a control series, in which the same type of grafting was done with normal embryos, replacement of posterior by anterior endoderm reduced the number of germ cells to almost zero, but replacement of anterior by posterior endoderm nearly doubled it. These findings are explained in terms of the distribution of the germ cells in the endoderm at the time of grafting. The results firstly show that the position of the germ cells is crucial to successful migration and secondly they support the notion that germ plasm has a determinative role during early germ cell differentiation.


Development ◽  
1961 ◽  
Vol 9 (3) ◽  
pp. 507-513
Author(s):  
Marie A. Di Berardino

The origin of the germ-cells has been extensively investigated in both invertebrates and vertebrates. In invertebrates it has been traced to the early cleavage of the zygote, e.g. in such forms as Ascaris megalocephala (Boveri, 1887) and Sciara (Metz, 1938). Within the vertebrate group the cases in which primordial germ-cells have been detected in cleavage stages are very few. Eigenmann (1891) identified gonocytes of Micrometrus aggregates in the late gastrula stage on the basis of their large size and the uniform distribution of their chromatin. The first clear demonstration of vertebrate primordial germ-cells appearing in a stage as early as the blastula was made in the European frog, Rana temporaria (Bounoure, 1934). In this case germ-cells were found to be conspicuous because of a stainable cytoplasmic element, the germ-plasm. This germ-plasm first appears shortly after fertilization in the form of islets concentrated in the vegetal pole region of the egg (Bounoure, 1934, 1939, 1954).


Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 251-265
Author(s):  
Yasuko Akita ◽  
Masami Wakahara

Correlation of the number of primordial germ cells (PGCs) at stage 47 with the amount of germ plasm at the 8-cell stage and with the number of the germ-plasm-containing cells (GPCCs) was analysed using two different laboratory-raised colonies of Xenopus laevis, HD and J groups. The average number of PGCs in J group tadpoles was significantly larger than that in HD group tadpoles. The amount of germ plasm in J group embryos was also demonstrated to be larger than in HD group embryos. The amount of germ plasm was related positively to the number of GPCCs at the 8-cell stage and to the resulting number of PGCs; embryos which contained larger amounts of germ plasm developed larger numbers of PGCs at stage 47. The average number of PGCs in experimentally induced triploid tadpoles was exactly twothirds of that in normal diploid tadpoles. Furthermore, in somatic cells (e.g. epidermis, muscle, pancreas), the number of cells in the triploid was also two-thirds of that in diploid tadpoles. These findings suggest that the number of PGCs is regulated by at least two different mechanisms: first, the number of PGCs is primarily specified by the intrinsic amount of germ plasm in the fertilized egg. Second, it is regulated by an unknown mechanism which controls the total number of cells of whole embryos, such as the nucleocytoplasmic ratio.


Development ◽  
2022 ◽  
Author(s):  
Yuki Naitou ◽  
Go Nagamatsu ◽  
Nobuhiko Hamazaki ◽  
Kenjiro Shirane ◽  
Masafumi Hayashi ◽  
...  

In mammals, primordial germ cells (PGCs), the origin of the germ line, are specified from the epiblast at the posterior region where gastrulation simultaneously occurs, yet the functional relationship between PGC specification and gastrulation remains unclear. Here, we show that Ovol2, a transcription factor conserved across the animal kingdom, balances these major developmental processes by repressing the epithelial-to-mesenchymal transition (EMT) driving gastrulation and the upregulation of genes associated with PGC specification. Ovol2a, a splice variant encoding a repressor domain, directly regulates EMT-related genes and consequently induces re-acquisition of potential pluripotency during PGC specification, whereas Ovol2b, another splice variant missing the repressor domain, directly upregulates genes associated with PGC specification. Taken together, these results elucidate the molecular mechanism underlying allocation of the germ line among epiblast cells differentiating into somatic cells through gastrulation.


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