Investigations of the Germ-plasm in Relation to Nuclear Transplantation

Development ◽  
1961 ◽  
Vol 9 (3) ◽  
pp. 507-513
Author(s):  
Marie A. Di Berardino
Keyword(s):  
Germ Cells ◽  
Germ Plasm ◽  
Rana Temporaria ◽  
Primordial Germ Cells ◽  
Nuclear Transplantation ◽  
Early Cleavage ◽  
Large Size ◽  
Vegetal Pole ◽  
Frog Rana Temporaria ◽  
Made In

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).

Partial characterization of ‘primordial germ cell-forming activity’ localized in vegetal pole cytoplasm in anuran eggs

Development ◽  
1977 ◽  
Vol 39 (1) ◽  
pp. 221-233
Author(s):  
Masami Wakahara
Keyword(s):  
Germ Cells ◽  
Germ Plasm ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Differential Centrifugation ◽  
Rana Chensinensis ◽  
Crude Homogenate ◽  
Germinal Granules ◽  
Vegetal Pole

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.

scholarly journals Transfer of Primordial Germ-cells between Two Subspecies of Xenopus laevis

Development ◽  
1962 ◽  
Vol 10 (4) ◽  
pp. 641-651
Author(s):  
A. W. Blackler
Keyword(s):  
Xenopus Laevis ◽  
Germ Cells ◽  
Nuclear Membrane ◽  
Rana Temporaria ◽  
Primordial Germ Cells ◽  
Staining Technique ◽  
Tail Bud ◽  
Ventral Region ◽  
Large Size ◽  
The Poor

In Anura the primordial germ-cells are discernible in the dorsal crest endoderm of tail-bud stages of development and may be traced from this position throughout their migration into the undifferentiated gonadal rudiment. These facts have been established by the descriptive studies of a number of workers (see review by Johnston, 1951), the cells being recognizable by their large size, the retention of yolk platelets long after their disappearance in neighbouring cells, the sharply denned and often kidney-shaped nuclear membrane, and the poor staining affinity of the nuclear contents. By means of the application of the Altmann-Volkonsky staining technique, Bounoure (1934) was able to demonstrate that germ-cells of the dorsal crest endoderm are the lineal descendants of certain cells found in the ventral region of the blastula. This discovery has been confirmed for Rana temporaria (the species investigated by Bounoure) by Blackler (1958), and extended to other Anuran species by Nieuwkoop (1956 a, b), Blackler (1958), and Di Berardino (1961).

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
Keyword(s):  
Xenopus Laevis ◽  
Germ Cells ◽  
Germ Plasm ◽  
Primordial Germ Cells ◽  
Cell Lineage ◽  
Initial Effect ◽  
Vegetal Pole ◽  
Cortical Regions ◽  
The Individual

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.

Germ plasm and the origin of the primordial germ cells in the oriental river prawn Macrobrachium nipponense

2021 ◽  
Author(s):  
Ying Chen ◽  
Xiang Fang ◽  
Xiao-Qing Tian ◽  
Zheng Cui ◽  
Hai-Yang Feng ◽  
...  
Keyword(s):  
Germ Cells ◽  
Germ Plasm ◽  
Primordial Germ Cells ◽  
Macrobrachium Nipponense ◽  
Oriental River Prawn

The effect of egg rotation on the differentiation of primordial germ cells in Xenopus laevis

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 79-99
Author(s):  
J. H. Cleine ◽  
K. E. Dixon
Keyword(s):  
Xenopus Laevis ◽  
Germ Cells ◽  
Germ Plasm ◽  
Primordial Germ Cells ◽  
Intermediate Zone ◽  
Gastrula Stage ◽  
Tail Bud ◽  
Axis Orientation ◽  
Early Gastrula ◽  
Number Of Cells

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.

The positional effect of presumptive primordial germ cells (pPGCs) on their differentiation into PGCs in Xenopus

Development ◽  
1988 ◽  
Vol 102 (3) ◽  
pp. 527-535
Author(s):  
K. Ikenishi ◽  
Y. Tsuzaki
Keyword(s):  
Germ Cells ◽  
Germ Plasm ◽  
Germ Line ◽  
Anterior Part ◽  
Primordial Germ Cells ◽  
Positional Effect ◽  
In Series

To determine whether the location of ‘germ plasm’-bearing cells [presumptive primordial germ cells (pPGCs)] is crucial for their differentiation into PGCs in Xenopus, [3H]thymidine-labelled pPGCs were implanted into the anterior or posterior halves of the endoderm in unlabelled host neurulae. Labelled PGCs in the genital ridges of experimental tadpoles were investigated by autoradiography. When the labelled pPGCs were implanted into posterior halves of the endoderm where host pPGCs are situated, 65 and 77% of the experimental tadpoles (designated as p-tadpoles) had the labelled PGCs in series I and II, respectively. When implanted into the anterior halves, 20 and 27% of the experimental tadpoles (a- tadpoles) had the labelled PGCs in series I and II, respectively. In p-tadpoles, the average numbers of labelled PGCs per tadpole were 8á7 in series I and 10 in series II, whereas they were 2á0 in a-tadpoles of both series. Both the proportion and the average number in p-tadpoles of both series were significantly different from those in a-tadpoles. In both series, labelled PGCs in p-tadpoles were found to be distributed throughout the genital ridges while those in a-tadpoles were localized only in the anterior part of the ridges. These facts indicate that the location of pPGCs in the endoderm affects their successful migration into the genital ridges, and that not only the presence of the germ plasm but also the proper location in endoderm are prerequisites to PGC differentiation of the germ line cells.

Nuclear transplantation of male primordial germ cells in the mouse

Development ◽  
1989 ◽  
Vol 107 (2) ◽  
pp. 407-411 ◽  
Author(s):  
Y. Tsunoda ◽  
T. Tokunaga ◽  
H. Imai ◽  
T. Uchida
Keyword(s):  
Germ Cells ◽  
Inner Cell Mass ◽  
Primordial Germ Cells ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Nuclear Transplantation ◽  
Embryonic Genome ◽  
Donor Nucleus ◽  
Implantation Sites ◽  
Genome Activation

We examined the developmental ability of enucleated eggs receiving embryonic nuclei and male primordial germ cells (PGCs) in the mouse. Reconstituted eggs developed into the blastocyst stage only when an earlier 2-cell nucleus was transplanted (36%) but very rarely if the donor nucleus was derived from a later 2-cell, 8-cell, or inner cell mass of a blastocyst (0–3%). 54–100%, 11–67%, 6–43% and 6–20% of enucleated eggs receiving male PGCs developed to 2-cell, 4-cell, 8-cell and blastocyst stage, respectively, in culture. The overall success rate when taking into account the total number of attempts at introducing germ cells was actually 0–6%. Live fetuses were not obtained after transfer of reconstituted eggs to recipients, although implantation sites were observed. The developmental ability of reconstituted eggs in relation to embryonic genome activation and genomic imprinting is discussed.

The Fine Structure of Primordial Germ Cells of the Rat

1973 ◽  
Vol 31 ◽  
pp. 634-635
Author(s):  
E. M. Eddy
Keyword(s):  
Endoplasmic Reticulum ◽  
Life History ◽  
Fine Structure ◽  
Rough Endoplasmic Reticulum ◽  
Germ Cells ◽  
Fibrous Material ◽  
Primordial Germ Cells ◽  
The Other ◽  
Large Size ◽  
History Of

Primordial germ cells are readily recognizable in embryos of the rat due to their large size, generally rounded shape and prominent nuclei with uniformly dispersed heterochromatin. They often have blunted pseudopodal processes at one end and small ruffles or trailing processes at the other, characteristics expected from their known ameboid activity- and migratory abilities. Also, the cytoplasm is rich in polyribosomes and contains a modest amount of rough endoplasmic reticulum and the mitochondria are frequently larger and less dense than those of adjacent somatic cells.In addition to these general characteristics, there are features unique to germ cells which allow them to be identified with certainty. These are: 1) small vesicles containing an irregular, dense core and 2) discrete accumulations of fibrous material known as nuage. Both of these features are present in other species and at other times in the life history of germ cells. The dense-cored vesicles have been noted in fetal and early postnatal mouse oogonia and oocytes, and in hamster and rabbit oocytes.

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