Polar granules and pole cells in the embryo of Calliphora erythrocephala: ultrastructure and [3H]leucine labelling

Development ◽  
1980 ◽  
Vol 57 (1) ◽  
pp. 79-93
Author(s):  
Anders Lundquist ◽  
Hadar Emanuelsson
Keyword(s):  
Cell Formation ◽  
Electron Microscopic ◽  
Electron Microscopic Autoradiography ◽  
Calliphora Erythrocephala ◽  
Pole Cell ◽  
Polar Granules ◽  
Pole Cells ◽  
Autoradiographic Analysis ◽  
Pole Plasm ◽  
Blastoderm Stage

The polar granules in Calliphora undergo a gradual fragmentation during early cleavage, but reaggregate after pole-cell formation. Autoradiographic analysis showed that the pole cells in Calliphora acquire a higher ‘3H’leucine label than the rest of the embryo during the blastoderm stage. Such an increased label was not seen in the pole plasm before pole-cell formation or in the pole cells during gastrulation. Electron microscopic autoradiography revealed that the polar granules are substantially labelled during the blastoderm stage. At the same time, characteristic nuclear blebs appear in the pole cells. The observations are consistent with the hypothesis that polar granules contain maternalmessenger RNA, which is released and translated into proteins.

scholarly journals Transcriptomic and functional analysis of the oosome, a unique form of germ plasm in the waspNasonia vitripennis

2018 ◽  
Author(s):  
Honghu Quan ◽  
Jeremy Lynch
Keyword(s):  
Molecular Basis ◽  
Germ Plasm ◽  
Cell Formation ◽  
Solid Sphere ◽  
The Novel ◽  
Developmental Networks ◽  
Pole Cell ◽  
Polar Granules ◽  
Developmental Role ◽  
Pole Cells

AbstractBackgroundThe oosome is the germline determinant in the waspNasonia vitripennisand is homologous to the polar granules ofDrosophila. Despite a common evolutionary origin and developmental role, the oosome is morphologically quite distinct from polar granules. It is a solid sphere that migrates within the cytoplasm before budding out and forming pole cells.ResultsTo gain an understanding of both the molecular basis of the novel form of the oosome, and the conserved essential features of germ plasm, we quantified and compared transcript levels between embryo fragments that contained the oosome, and those that did not. The identity of the localized transcripts indicated thatNasoniauses different molecules to carry out conserved germ plasm functions. In addition, functional testing of a sample of localized transcripts revealed potentially novel mechanisms of ribonucleoprotein assembly and pole cell cellularization in the wasp.ConclusionsOur results demonstrate that numerous novel and unexpected molecules have been recruited in order produce the unique characteristics of the oosome and pole cell formation inNasonia. This work will serve as the basis for further investigation into the patterns of germline determinant evolution among insects, the molecular basis of extreme morphology of ribonucleoproteins, and the incorporation of novel components into developmental networks.

Restoration of pole-cell-forming ability to u.v.-irradiated Drosophila embryos by injection of mitochondrial lrRNA

Development ◽  
1989 ◽  
Vol 107 (4) ◽  
pp. 733-742 ◽  
Author(s):  
S. Kobayashi ◽  
M. Okada
Keyword(s):  
Dna Sequences ◽  
Nuclear Dna ◽  
Mitochondrial Gene ◽  
Cell Formation ◽  
Nuclear Bodies ◽  
Cdna Insert ◽  
Pole Cell ◽  
Polar Granules ◽  
Pole Cells

Screening a cDNA library generated from poly(A) +RNA of Drosophila cleavage embryos, we selected a cDNA clone (pDE20.6). The cDNA hybridized specifically with a poly(A) +RNA that is capable of restoring embryos from u.v.-caused inability of pole cell formation. The RNA hybrid-selected by pDE20.6 was also able to induce pole cells in the anterior region of embryos, if it was coinjected with u.v.-irradiated polar plasm, although the RNA or irradiated polar plasm alone was not effective. Pole cells thus formed in the anterior or in the u.v.-irradiated posterior region were identified by polar granules and nuclear bodies, morphological markers for normal pole cells. Furthermore, the RNA-induced pole cells were able to migrate into gonadal rudiments. The nucleotide sequence of pDE20.6 cDNA insert was highly homologous with the mitochondrial large rRNA (lrRNA) gene, but not with any nuclear DNA sequences. Using pDE20.6 as a primer, a full-length cDNA of mitochondrial lrRNA was generated and cloned. The RNA transcribed in vitro from the cDNA was able to restore pole cell formation. The cDNA hybridized only with a 1.5 kb poly(A) +RNA on a Northern blot. The 1.5 kb RNA sedimented more with the post-mitochondrial (P3) fraction than with the mitochondrial (P2) fraction, while the majority of transcripts from another mitochondrial gene was detected in the P2 fraction.

Spatial and developmental changes in the respiratory activity of mitochondria in early Drosophila embryos

Development ◽  
1992 ◽  
Vol 115 (4) ◽  
pp. 1175-1182 ◽  
Author(s):  
T. Akiyama ◽  
M. Okada
Keyword(s):  
Respiratory Activity ◽  
Cell Formation ◽  
Rhodamine 123 ◽  
Mitochondrial Activity ◽  
Early Cleavage ◽  
Pole Cell ◽  
Posterior Group ◽  
Cleavage Stages ◽  
Pole Cells ◽  
Drosophila Embryos

Mitochondria of early Drosophila embryos were observed with a transmission electron microscope and a fluorescent microscope after vital staining with rhodamine 123, which accumulates only in active mitochondria. Rhodamine 123 accumulated particularly in the posterior pole region in early cleavage embryos, whereas the spatial distribution of mitochondria in an embryo was uniform throughout cleavage stages. In late cleavage stages, the dye showed very weak and uniform accumulation in all regions of periplasm. Polar plasm, sequestered in pole cells, restored the ability to accumulate the dye. Therefore, it is concluded that the respiratory activity of mitochondria is higher in the polar plasm than in the other regions of periplasm in early embryos, and this changes during development. The temporal changes in rhodamine 123-staining of polar plasm were not affected by u.v. irradiation at the posterior of early cleavage embryos at a sufficient dosage to prevent pole cell formation. This suggests that the inhibition of pole cell formation by u.v. irradiation is not due to the inactivation of the respiratory activities of mitochondria. In addition, we found that the anterior of Bicaudal-D mutant embryos at cleavage stage was stained with rhodamine 123 with the same intensity as the posterior of wild-type embryos. No pole cells form in the anterior of Bic-D embryos, where no restoration of mitochondrial activity occurs in the blastoderm stage. The posterior group mutations that we tested (staufen, oskar, tudor, nanos) and the terminal mutation (torso) did not alter staining pattern of the posterior with rhodamine 123.

Differentiation and positioning of nuclei in eggs of the cecidomyid Heteropeza pygmaea

1976 ◽  
Vol 22 (1) ◽  
pp. 99-113
Author(s):  
M. Meats ◽  
J.B. Tucker
Keyword(s):  
Structural Changes ◽  
Early Stage ◽  
Cell Formation ◽  
Longitudinal Axis ◽  
Posterior Pole ◽  
Spindle Orientation ◽  
Specific Sequence ◽  
Pole Cell ◽  
Polar Granules ◽  
Egg Chamber

During the first three cleavage divisions of the egg nuclei a precise sequence of spindle orientation and elongation parallel to the longitudinal axis of the egg is apparently involved in positioning one nucleus among the polar granules at the posterior pole of the egg. The size of this nucleus, and the position at which the egg cleaves when pole cell formation occurs, appear to constitute part of the mechanism which ensures that only one nucleus is included in the first pole cell. Blastoderm formation occurs without a well-defined migration of nuclei to the egg surface. Nuclei are so large in relation to the size of the egg that uniform spacing and distribution of nuclei ensures that a large proportion are situated near the egg surface. Those nuclei which are near the egg surface divide synchronously to form a layer of blastoderm nuclei, while membranous cleavage furrows invaginate from the egg surface between them. Nuclei in the central region of the egg chamber condense to form yolk nuclei before blastoderm nuclei have been separated from the rest of the egg by the completion of the cleavage membranes. Polar granules provide the only evidence of fine-structural differences in different regions of the egg chamber cytoplasm. They are found near the posterior pole of the egg from an early stage of oogenesis. They undergo a specific sequence of structural changes and increase in size as the egg grows. No microtubular or microfibrillar arrays have been found in the egg chamber which might form a cytoskeletal basis for spindle orientation or for the spatial differences which develop during differentiation of the uncleaved egg cytoplasm.

scholarly journals Identification of a component of Drosophila polar granules

Development ◽  
1988 ◽  
Vol 103 (4) ◽  
pp. 625-640 ◽  
Author(s):  
B. Hay ◽  
L. Ackerman ◽  
S. Barbel ◽  
L.Y. Jan ◽  
Y.N. Jan
Keyword(s):  
Germ Line ◽  
Posterior Pole ◽  
Nuclear Bodies ◽  
Maternal Origin ◽  
Pole Cell ◽  
Polar Granules ◽  
Early Embryos ◽  
Biochemical Studies ◽  
Males And Females ◽  
Pole Cells

Information necessary for the formation of pole cells, precursors of the germ line, is provided maternally and localized to the posterior pole of the Drosophila egg. The maternal origin and posterior localization of polar granules suggest that they may be associated with pole cell determinants. We have generated an antibody (Mab46F11) against polar granules. In oocytes and early embryos, the Mab46F11 antigen is sharply localized to the posterior embryonic pole. In pole cells, it becomes associated with nuclear bodies within, and nuage around, the nucleus. Immunoreactivity remains associated with cells of the germ line throughout the life cycle of both males and females. This antibody recognizes a 72–74 × 10(3) Mr protein and is useful both as a pole lineage marker and in biochemical studies of polar granules.

Restoration of the capacity to form pole cells in u.v.-irradiated Drosophila embryos

Development ◽  
1975 ◽  
Vol 33 (4) ◽  
pp. 1003-1011
Author(s):  
Richard Warn
Keyword(s):  
Pole Cell ◽  
Pole Cells ◽  
Pole Plasm ◽  
Cell Fragments ◽  
Drosophila Embryos

Injection of pole plasm into u.v.-irradiated posterior poles of early Drosophila embryos leads to the restoration of the capacity to form pole cells in nearly half of the recipients. The effect is specific, since cytoplasm from the anterior tip has no such result. In most cases only a small number (between 1 and 5) of discrete pole cells are formed. However, a large number of pole cell fragments with or without nuclei occur. Occasionally pole cells were formed outside the area of the originally irradiated pole plasm. This happened when material was injected more anteriorly than usual. Thus polar cytoplasm contains some factor(s) necessary for the formation of pole cells.

Oskar anchoring restricts pole plasm formation to the posterior of the Drosophila oocyte

Development ◽  
2002 ◽  
Vol 129 (15) ◽  
pp. 3705-3714 ◽  
Author(s):  
Nathalie F. Vanzo ◽  
Anne Ephrussi
Keyword(s):  
Drosophila Melanogaster ◽  
Translational Control ◽  
Cell Formation ◽  
Positional Information ◽  
Mrna Localization ◽  
Posterior Pole ◽  
Tight Coupling ◽  
Anteroposterior Patterning ◽  
Pole Cell ◽  
Pole Plasm

Localization of the maternal determinant Oskar at the posterior pole of Drosophila melanogaster oocyte provides the positional information for pole plasm formation. Spatial control of Oskar expression is achieved through the tight coupling of mRNA localization to translational control, such that only posterior-localized oskar mRNA is translated, producing the two Oskar isoforms Long Osk and Short Osk. We present evidence that this coupling is not sufficient to restrict Oskar to the posterior pole of the oocyte. We show that Long Osk anchors both oskar mRNA and Short Osk, the isoform active in pole plasm assembly, at the posterior pole. In the absence of anchoring by Long Osk, Short Osk disperses into the bulk cytoplasm during late oogenesis, impairing pole cell formation in the embryo. In addition, the pool of untethered Short Osk causes anteroposterior patterning defects, owing to the dispersion of pole plasm and its abdomen-inducing activity throughout the oocyte. We show that the N-terminal extension of Long Osk is necessary but not sufficient for posterior anchoring, arguing for multiple docking elements in Oskar. This study reveals cortical anchoring of the posterior determinant Oskar as a crucial step in pole plasm assembly and restriction, required for proper development of Drosophila melanogaster.

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 Three distinct distributions of F-actin occur during the divisions of polar surface caps to produce pole cells in Drosophila embryos.

1985 ◽  
Vol 100 (4) ◽  
pp. 1010-1015 ◽  
Author(s):  
R M Warn ◽  
L Smith ◽  
A Warn
Keyword(s):  
Cell Formation ◽  
Contractile Ring ◽  
Polar Surface ◽  
Ring Type ◽  
Actin Organization ◽  
Polar Caps ◽  
Pole Cell ◽  
Nuclear Cycle ◽  
Pole Cells ◽  
Drosophila Embryos

The F-actin distribution was studied during pole cell formation in Drosophila embryos using the phalloidin derivative rhodaminyl-lysine-phallotoxin. Nuclei were also stained with 4'-6 diamidine-2-phenylindole dihydrochloride to correlate the pattern seen with the nuclear cycle. The precursors of the pole cells, the polar surface caps, were found to have an F-actin-rich cortex distinct from that of the rest of the embryo surface and an interior cytoplasm that was less intensely stained but brighter than the cytoplasm deeper in the embryo. They were found to divide once without forming true cells and then a second time when cells formed as a result of a meridional and a basal cleavage. Three distinct distributions of the cortical F-actin have been identified during these cleavages. It is concluded that the first division, which cleaves the polar caps but does not separate them from the embryo, involves very different processes from those that lead to the formation of the pole cells. A contractile-ring type of F-actin organization may not be present during the first cleavage but is suggested to occur during the second.

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