Distribution of tudor protein in the Drosophila embryo suggests separation of functions based on site of localization

Development ◽  
1993 ◽  
Vol 119 (1) ◽  
pp. 207-219 ◽  
Author(s):  
A. Bardsley ◽  
K. McDonald ◽  
R.E. Boswell
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Germ Plasm ◽  
Primordial Germ Cells ◽  
Posterior Pole ◽  
Drosophila Embryo ◽  
Normal Functioning ◽  
Polar Granules ◽  
Pole Cells

Mutations in the tudor locus of Drosophila affect two distinct determinative processes in embryogenesis; segmentation of the abdomen and determination of the primordial germ cells. The distribution of tudor protein during embryogenesis, and the effect of various mutations on its distribution, suggest that tudor protein may carry out these functions separately, based on its location in the embryo. The protein is concentrated in the posterior pole cytoplasm (germ plasm), where it is found in polar granules and mitochondria. Throughout the rest of the embryo, tudor protein is associated with the cleavage nuclei. Mutations in all maternal genes known to be required for the normal functioning of the germ plasm eliminate the posterior localization of tudor protein, whereas mutations in genes required for the functioning of the abdominal determinant disrupt the localization around nuclei. Analysis of embryos of different maternal genotypes indicates that the average number of pole cells formed is correlated with the amount of tudor protein that accumulates in the germ plasm. Our results suggest that tudor protein localized in the germ plasm is instrumental in germ cell determination, whereas nuclear-associated tudor protein is involved in determination of segmental pattern in the abdomen.

Epsilon granules in Drosophila pole cells and oöcytes

Development ◽  
1965 ◽  
Vol 13 (1) ◽  
pp. 73-81
Author(s):  
Suzanne L. Ullmann
Keyword(s):  
Germ Cells ◽  
Microscope Study ◽  
Light Microscope ◽  
Primordial Germ Cells ◽  
Structural Level ◽  
Developmental Morphology ◽  
Insect Eggs ◽  
Polar Granules ◽  
Pole Cells ◽  
Drosophila Embryos

In many insect eggs, including those of the Diptera, deeply staining granules, rich in RNA, occur in the posterior polar plasm and during ontogeny become enclosed within the pole cells. The structure and fate of these cells, which generally give rise to the primordial germ cells, and their inclusions have excited interest for over half a century (Hegner, 1908; Huettner, 1923; Rabinowitz, 1941; Poulson, 1947; Counce, 1963; Mahowald, 1962), yet numerous questions concerning them remain unsettled or controversial to this day. For instance, the dual fate of the pole cells in Drosophila, the genus which has been most extensively studied, is still debated (Poulson & Waterhouse, 1960; Hathaway & Selman, 1961). Recently, Counce (1963), in a light-microscope study, has described the developmental morphology of the polar granules in several species of Drosophila embryos; while Mahowald (1962) has succeeded in identifying them in D. melanogaster at the ultra-structural level.

Experimental studies on the role of ‘polar granules’ in the segregation of pole cells in Drosophila melanogaster

Development ◽  
1966 ◽  
Vol 16 (3) ◽  
pp. 391-399
Author(s):  
Bożenna Jazdowska-Zagrodzińska
Keyword(s):  
Germ Cells ◽  
Normal Development ◽  
Primordial Germ Cells ◽  
Experimental Studies ◽  
Main Body ◽  
Track Formation ◽  
Polar Granules ◽  
Pole Cells ◽  
Pole Plasm

The early differentiation of germ cells is a common phenomenon in the animal kingdom. Insects are of special interest in this respect, as the differentiation of their primordial germ cells occurs in very early stages of cleavage (Kahle, 1908; Hegner, 1914; Reitberger, 1934; Kraczkiewicz, 1935, 1936) and the structure of the ooplasm enables relatively convenient observation of the phenomenon of germ track formation. The ooplasm is differentiated in that the posterior end of the egg contains the so-called ‘pole plasm’ in which there are easily visible inclusions quite different from yolk, though staining similarly with haematoxylin. Such inclusions are not noted in other parts of the egg. In the course of normal development the region containing granules and pole plasm always detaches, producing the primordial germ cells. During the separation of the primordial germ cells, also called pole cells, all these granules become included in their cytoplasm, and the main body of ooplasm is left devoid of them.

scholarly journals Two types of pole cells are present in the Drosophila embryo, one with and one without splicing activity for the third P-element intron

Development ◽  
1993 ◽  
Vol 117 (3) ◽  
pp. 885-893 ◽  
Author(s):  
S. Kobayashi ◽  
T. Kitamura ◽  
H. Sasaki ◽  
M. Okada
Keyword(s):  
Germ Cells ◽  
Germ Line ◽  
Primordial Germ Cells ◽  
Egg Laying ◽  
P Element ◽  
Drosophila Embryo ◽  
The Third ◽  
Pole Cells ◽  
Almost All ◽  
Second Peak

In Drosophila, it has been postulated that the third intron of the P-element is spliced only in germ-line cells. To test whether this postulate is applicable to pole cells, the progenitor cells of germ line, we carried out a histochemical assay to detect the splicing activity in embryos. The splicing activity was detected in pole cells and primordial germ cells. The activity increased to reach a maximum at 5–6 hours AEL (after egg laying), then decreased to an undetectable level by 8–9 hours AEL. The splicing activity showed a small second peak at 12–15 hours AEL. It was rather unexpected that not all pole cells were capable of splicing the third intron. Almost all pole cells that had the splicing activity at 5–6 hours AEL penetrated the embryonic gonads and differentiated into primordial germ cells. Our findings suggest that pole cells are selected to penetrate the gonads while they are migrating from the proctodeal cavity to the gonads. Furthermore, these results suggest that the machinery to splice the P-element is active in some pole cells, and that this activity is used for processing transcripts of genes that play important roles in the differentiation of pole cells into primordial germ cells.

Experiments on chromosome elimination in the gall midge, Mayetiola destructor

Development ◽  
1970 ◽  
Vol 24 (2) ◽  
pp. 257-286
Author(s):  
C. R. Bantock
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Germ Line ◽  
Gall Midge ◽  
Primordial Germ Cells ◽  
Chromosome Complement ◽  
Chromosome Elimination ◽  
Mayetiola Destructor ◽  
Cell Nuclei ◽  
Polar Granules

Cleavage in Cecidomyidae (Diptera) is characterized by the elimination of chromosomes from presumptive somatic nuclei. The full chromosome complement is kept by the germ-line nuclei. The course of cleavage in Mayetiola destructor (Say) is described. After the fourth division two nuclei lie in the posterior polar-plasm and become associated with polar granules, and fourteen nuclei lie in the rest of the cytoplasm. All the nuclei possess about forty chromosomes. During the fifth division the posterior nuclei do not divide and the polar-plasm becomes constricted to form primordial germ cells (pole cells). The remaining fourteen nuclei divide and lose about thirty-two chromosomes so that twenty-eight nuclei are formed containing only eight chromosomes. These are the presumptive somatic nuclei. During subsequent divisions the pole cell nuclei retain the full chromosome number; these divisions occur less frequently than those of the somatic nuclei. Experiments were performed on early embryonic stages to elucidate the properties of the posterior end during the time that chromosome elimination was taking place from the presumptive somatic nuclei. Ultraviolet irradiation, constriction, and centrifugation techniques were used. The polar granules are concerned with the non-division of the germ-cell nuclei during the fifth division, since if the granules are dispersed by centrifugation, or if nuclei are prevented by constriction from coming into contact with them before the fifth division, all the nuclei divide with chromosome elimination at this division. With each technique it is possible to obtain embryos possessing germ cells with only eight chromosomes in their nuclei. Individuals possessing germ-line nuclei with only eight chromosomes were allowed to develop to maturity. Abnormalities were confined to the germ cells only and were the same regardless of which technique had been used to produce the deficient germ line. An ovary containing germ-cell nuclei with only eight chromosomes is unable to form both oocytes and nurse cells. A testis containing germ-cell nuclei with only eight chromosomes is unable to form spermatocytes but cells which come to resemble gametes are formed. Experimental males and females are both sterile. The results are discussed in relation to other experimental work on Cecidomyidae and the following main conclusions are reached: (a) the polar granules are responsible for preventing an irreversible loss of chromosomes from the germ-cell nuclei by preventing the mitosis of these nuclei during the fifth division; (b) the chromosomes normally retained in the germ line are required for gametogenesis, particularly for oogenesis. The significance of chromosome elimination is discussed.

Replacement of posterior by anterior endoderm reduces sterility in embryos from inverted eggs of Xenopus laevis

Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 83-93
Author(s):  
J. H. Cleine
Keyword(s):  
Cell Differentiation ◽  
Xenopus Laevis ◽  
Germ Cell ◽  
Germ Cells ◽  
Germ Plasm ◽  
Primordial Germ Cells ◽  
Inverted Position ◽  
Control Series ◽  
Germ Cell Differentiation ◽  
Tailbud Stage

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.

scholarly journals Phase transitioned nuclear Oskar promotes cell division of Drosophila primordial germ cells

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Kathryn E Kistler ◽  
Tatjana Trcek ◽  
Thomas R Hurd ◽  
Ruoyu Chen ◽  
Feng-Xia Liang ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Fate ◽  
Germ Cells ◽  
Cell Function ◽  
Primordial Germ Cells ◽  
Drosophila Embryo ◽  
Nuclear Localization Sequence ◽  
Cell Systems ◽  
Germ Granules ◽  
Localization Sequence

Germ granules are non-membranous ribonucleoprotein granules deemed the hubs for post-transcriptional gene regulation and functionally linked to germ cell fate across species. Little is known about the physical properties of germ granules and how these relate to germ cell function. Here we study two types of germ granules in the Drosophila embryo: cytoplasmic germ granules that instruct primordial germ cells (PGCs) formation and nuclear germ granules within early PGCs with unknown function. We show that cytoplasmic and nuclear germ granules are phase transitioned condensates nucleated by Oskar protein that display liquid as well as hydrogel-like properties. Focusing on nuclear granules, we find that Oskar drives their formation in heterologous cell systems. Multiple, independent Oskar protein domains synergize to promote granule phase separation. Deletion of Oskar’s nuclear localization sequence specifically ablates nuclear granules in cell systems. In the embryo, nuclear germ granules promote germ cell divisions thereby increasing PGC number for the next generation.

scholarly journals Localization of vasa, a component of Drosophila polar granules, in maternal-effect mutants that alter embryonic anteroposterior polarity

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 425-433 ◽  
Author(s):  
B. Hay ◽  
L.Y. Jan ◽  
Y.N. Jan
Keyword(s):  
Germ Cell ◽  
Maternal Effect ◽  
Polar Granule ◽  
Cell Lineage ◽  
Posterior Pole ◽  
Drosophila Embryo ◽  
Polar Granules ◽  
Early Drosophila Embryo ◽  
Anteroposterior Polarity ◽  
Source Of Information

Cytoplasm at the posterior pole of the early Drosophila embryo, known as polar plasm, serves as a source of information necessary for germ cell determination and for specification of the abdominal region. Likely candidates for cytoplasmic elements important in one or both of these processes are polar granules, organelles concentrated in the cortical cytoplasm of the posterior pole. Females homozygous for any one of the maternal-effect mutations, tudor, oskar, staufen, vasa, or valois give rise to embryos that lack localized polar granules, fail to form the germ cell lineage and have abdominal segment deletions. Using antibodies against a polar granule component, the vasa protein, we find that vasa synthesis or localization is affected by these mutations. In vasa mutants, synthesis of vasa protein is absent or severely restricted. In oskar and staufen mutant females, vasa synthesis appears normal, but the vasa protein is not localized. In tudor and valois mutant females, vasa is localized to the posterior pole of oocytes, but this localization is lost following egg activation. In addition to the posterior localized vasa, there is a low level of vasa distributed throughout the embryo. A function for this distributed vasa is postulated based on the observation that embryos from Bicaudal-D mothers, in which abdominal determinants are incorrectly localized to the anterior pole, do not show any ectopic vasa localization, though abdomen development at the anterior end depends on the amount of vasa protein in the embryo.

scholarly journals Ligand–Receptor Interactions Elucidate Sex-Specific Pathways in the Trajectory From Primordial Germ Cells to Gonia During Human Development

2021 ◽  
Vol 9 ◽  
Author(s):  
Arend W. Overeem ◽  
Yolanda W. Chang ◽  
Jeroen Spruit ◽  
Celine M. Roelse ◽  
Susana M. Chuva De Sousa Lopes
Keyword(s):  
Germ Cell ◽  
Signaling Pathways ◽  
Germ Cells ◽  
Tyrosine Kinases ◽  
Intracellular Localization ◽  
Primordial Germ Cells ◽  
Cell Lineage ◽  
Systematic Analysis ◽  
Receptor Interactions

The human germ cell lineage originates from primordial germ cells (PGCs), which are specified at approximately the third week of development. Our understanding of the signaling pathways that control this event has significantly increased in recent years and that has enabled the generation of PGC-like cells (PGCLCs) from pluripotent stem cells in vitro. However, the signaling pathways that drive the transition of PGCs into gonia (prospermatogonia in males or premeiotic oogonia in females) remain unclear, and we are presently unable to mimic this step in vitro in the absence of gonadal tissue. Therefore, we have analyzed single-cell transcriptomics data of human fetal gonads to map the molecular interactions during the sex-specific transition from PGCs to gonia. The CellPhoneDB algorithm was used to identify significant ligand–receptor interactions between germ cells and their sex-specific neighboring gonadal somatic cells, focusing on four major signaling pathways WNT, NOTCH, TGFβ/BMP, and receptor tyrosine kinases (RTK). Subsequently, the expression and intracellular localization of key effectors for these pathways were validated in human fetal gonads by immunostaining. This approach provided a systematic analysis of the signaling environment in developing human gonads and revealed sex-specific signaling pathways during human premeiotic germ cell development. This work serves as a foundation to understand the transition from PGCs to premeiotic oogonia or prospermatogonia and identifies sex-specific signaling pathways that are of interest in the step-by-step reconstitution of human gametogenesis in vitro.

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