scholarly journals DNA SYNTHESIS IN THE OOPLASM OF DROSOPHILA MELANOGASTER

1966 ◽  
Vol 28 (2) ◽  
pp. 199-208 ◽  
Author(s):  
F. A. Muckenthaler ◽  
A. P. Mahowald

Tritiated thymidine was injected into 2-day-old Drosophila melanogaster females, and tissue sections were prepared from the ovary for radioautography with both the light and electron microscopes. Besides the expected incorporation of H3-thymidine into nuclei of nurse cells and follicle cells, there was a relatively high level of incorporation of label into ooplasmic DNA. The highest level of incorporation occurred at stage 12. At the same time, the 15 nurse cell nuclei also incorporate thymidine in spite of the fact that they are breaking down and degenerating. The label in the ooplasm is not removed by extraction with DNase (although this removes nuclear label) unless extraction is preceded by a treatment with protease. Electron microscopic radioautography revealed that 36% of the silver grains resulting from decay of H3-thymidine are found over mitochondria, with a further 28% being located within 0.25 µ of these organelles. The remaining 36% of the silver grains was not found to be associated with any organelles, and it probably represents synthesis in the cytoplasm by the "storage DNA" characteristic of many eggs. It is suggested that one mechanism acting throughout the egg chamber is responsible for the synchronous synthesis of DNA in the degenerating nurse cells, in the mitochondria of the egg, and in the "storage DNA" of the ooplasm.

2010 ◽  
Vol 190 (4) ◽  
pp. 523-531 ◽  
Author(s):  
Ioannis P. Nezis ◽  
Bhupendra V. Shravage ◽  
Antonia P. Sagona ◽  
Trond Lamark ◽  
Geir Bjørkøy ◽  
...  

Autophagy is an evolutionarily conserved pathway responsible for degradation of cytoplasmic material via the lysosome. Although autophagy has been reported to contribute to cell death, the underlying mechanisms remain largely unknown. In this study, we show that autophagy controls DNA fragmentation during late oogenesis in Drosophila melanogaster. Inhibition of autophagy by genetically removing the function of the autophagy genes atg1, atg13, and vps34 resulted in late stage egg chambers that contained persisting nurse cell nuclei without fragmented DNA and attenuation of caspase-3 cleavage. The Drosophila inhibitor of apoptosis (IAP) dBruce was found to colocalize with the autophagic marker GFP-Atg8a and accumulated in autophagy mutants. Nurse cells lacking Atg1 or Vps34 in addition to dBruce contained persisting nurse cell nuclei with fragmented DNA. This indicates that autophagic degradation of dBruce controls DNA fragmentation in nurse cells. Our results reveal autophagic degradation of an IAP as a novel mechanism of triggering cell death and thereby provide a mechanistic link between autophagy and cell death.


1977 ◽  
Vol 25 (7) ◽  
pp. 655-661 ◽  
Author(s):  
M Pfoch ◽  
W Kade

In classifying cells in tissue sections, one must consider the fact that only random projections of cells and of subcellular structures are available in the two-dimensional image. Therefore, measurement values that solely reflect the size of such projections cannot be taken on their own as a basis for cell classification. More complex morphologic features such as shape, texture and distribution pattern of cells and their components should be analyzed. Using cell nuclei as an example, the relationship between such features and geometric measurement values is evaluated. It can be shown that a well balanced combination of geometric parameters provides a suitable basis for reproducing the visual preclassification of lymphocytes in tissue sections. Moreover, using a cluster algorithm, which allows different levels of similarity to be defined, a hierarchical sequence of subclusters turns out, indicating the heterogeneity of the visually determined cell classes. Whether or not these subclusters can be correlated to functionally defined subpopulations of lymphocytes remains a matter for further investigation.


Development ◽  
1976 ◽  
Vol 35 (3) ◽  
pp. 521-533
Author(s):  
F. Giorgi ◽  
P. Deri

An ultrastructural analysis has been made of certain ovarian chambers undergoing ab-normal development. The earliest morphological change in these chambers consists of thealteration of the nuclear material which is then followed by engulfment of portions of thenurse cell cytoplasm, including the nuclear fragments, into the overlying follicle cells. Thecontinuation of this process leads to the progressive disappearance of nurse cells with theconcomitant formation of huge dense vacuoles in the follicle layer. The morphological featuresdescribed in the present investigation are similar to those found in other tissues and inter-preted as leading to cell death. It is suggested that certain ovarian chambers undergo celldeath as a result of the incapability of furthering their development. The role played by celldeath in oogenesis is also discussed on the basis of the current literature.


1930 ◽  
Vol s2-73 (292) ◽  
pp. 617-630
Author(s):  
R.A. R. GRESSON

1. By the use of Feulgen's ‘nuclealreaktion’ certain points of Tenthredinid oogenesis have been subjected to closer study. The chromatin of the early nurse-cells of Allantus pallipes exists in the form of granules, the majority of which occur close to the nuclear membrane. In the older cells a nuclear network appears in which is distributed granules of chromatin. In Thrinax mixta, where the ovarioles were more highly developed, the chromatin of the nurse-cells occurs as granules scattered through the nucleus; a nuclear network is not present, but certain granules appear to be connected by a thread. The granules which were shown to surround the nurse-cell nuclei (in material treated by Bensley's method and also by fixation in Bouin's picro-formol and subsequently stained in iron haematoxylin) and which were formerly regarded as chromatin emissions from the nurse-cell nuclei (9) were not revealed by Feulgen's technique. They therefore cannot be regarded as chromatin. Their precise nature and origin remains undetermined. 2. The nucleoli of the early nurse-cells of both species, as revealed by Mann's methyl-blue eosin, are faintly basophil. Later they break up into a number of basophil bodies which undergo fragmentation; formerly (technique and reference as in 1) the basophil nucleolus and the basophil bodies originating from it were termed ‘nuclear material’ undergoing fragmentation. While this basophil nucleolar material presents a fragmented appearance, it increases in amount as evidenced by the large number of basophil bodies present in the older nurse-cell nuclei. This material is utilized for the nourishment of the egg after the latter engulfs the nurse-cell nuclei. Nucleolar extrusions to the cytoplasm do not occur. 3. The behaviour of the chromatin of the follicle-cell nuclei is similar to that of the nurse-cell nuclei except that in Allantus pallipes the nuclear chromatin network as demonstrated by Feulgen's technique disappears in the older cells. 4. The nucleoli of the follicle-cells are basophil. They become broken up in the older cells, but in most cases the resulting masses remain in contact. Nucleolar extrusions to the cytoplasm do not occur. 5. The occurrence of deeply basophil material in the cytoplasm of the follicle-cells of Thrinax mixta stained with Mann's methyl-blue eosin, formerly described for Bouin fixed material stained in iron haematoxylin (9), suggests that some substance in solution may be passed into the ooplasm; extrusion of granules from the follicle-cells to the ooplasm does not take place. 6. The absence or non-visibility of chromatin (Feulgen's technique) from the oocytes of Thrinax mixta, and its disappearance from the older oocytes of Allantus pallipes , would indicate that the chromatin undergoes a chemical change during oogenesis such as suggested by Koch for Chilopods. 7. The oxyphil and basophil nucleoli of the oocytes do not react to Feulgen's technique for chromatin; this agrees with Ludford's findings for the mouse and for Limnaeastagnalis.


Author(s):  
J. E. Michaels ◽  
J. T. Hung ◽  
E. L. Cardell ◽  
R. R. Cardell

In order to study early events of glycogen synthesis, we have used adrenalectomized (ADX) rats fasted overnight and injected with the synthetic glucocorticoid dexamethasone (DEX) to stimulate glycogen synthesis. Rats were given DEX 0-5 hr prior to sacrifice and injected with 2 mCi 3H-galactose 1 hr prior to sacrifice. Liver was prepared for light (LM) and electron microscopic (EM) radioautography by routine procedures.The concentration of silver grains over hepatic cytoplasm was measured in LM radioautographs using a Zeiss Videoplan. The hepatocytes were categorized as unlabeled if no silver grains (gr) were present, lightly labeled (<10gr/100 μm2 cytoplasm) or intensely labeled (>10 gr/1002 μm cytoplasm). Although very few hepatocytes showed heavy labeling after 1 hr treatment with DEX, by 2 hr after DEX treatment 8% of the cells distributed throughout the lobule were intensely labeled.


Author(s):  
Seiji Shioda ◽  
Yasumitsu Nakai ◽  
Atsushi Ichikawa ◽  
Hidehiko Ochiai ◽  
Nobuko Naito

The ultrastructure of neurosecretory cells and glia cells in the supraoptic nucleus (SON) of the hypothalamus and the neurohypophysis (PN) was studied after rapid freezing followed by substituion fixation. Also, the ultrastructural localization of vasopressin (VP) or its carrier protein neurophys in II (NPII) in the SON and PN was demonstrated by using a post-embedding immunoco1loidal gold staining method on the tissue sections processed by rapid freezing and freeze-substitution fixation.Adult male Wistar rat hypothalamus and pituitary gland were quenched by smashing against a copper block surface precooled with liquid helium and freeze-substituted in 3% osmium tetroxide-acetone solutions kept at -80°C for 36-48h. After substituion fixation, the tissue blocks were warmed up to room temperature, washed in acetone and then embedded in an Epon-Araldite mixture. Ultrathin sections mounted on 200 mesh nickel grids were immersed in saturated sodium metaperiodate and then incubated in each of the following solutions: 1 % egg albumin in phosphate buffer, VP or NPII (1/1000-1/5000) antiserum 24h at 4°C, 3) colloidal gold solution (1/20) 1h at 20°C. The sections were washed with distilled waterand dried, then stained with uranylacetate and lead citrate and examined with Hitachi HU-12A and H-800 electron microscopes.


2020 ◽  
Vol 62 (1-2) ◽  
pp. 151-161
Author(s):  
T. Shagholi ◽  
M. Keshavarzi ◽  
M. Sheidai

Tamarix L. (Tamaricaceae) is a halophytic shrub in different parts of Asia and North Africa. Taxonomy and species limitation of Tamarix is very complex. This genus has three sections as Tamarix, Oligadenia, and Polyadenia, which are mainly separated by petal length, the number of stamens, the shape of androecial disk and attachment of filament on the androecial disk. As there was no palynological data on pollen features of Tamarix species of Iran, in the present study 12 qualitative and quantitative pollen features were evaluated to find diagnostic ones. Pollen grains of 8 Tamarix species were collected from nature. Pollen grains were studied without any treatment. Measurements were based on at least 50 pollen grains per specimen. Light and scanning electron microscopes were used. Multivariate statistical methods were applied to clarify the species relationships based on pollen data. All species studied showed monad and tricolpate (except some individuals of T. androssowii). Some Tamarix species show a high level of variability, in response to ecological niches and phenotypic plasticity, which make Tamarix species separation much more difficult. Based on the results of the present study, pollen grains features are not in agreement with previous morphological and molecular genetics about the sectional distinction.


Genetics ◽  
1997 ◽  
Vol 146 (1) ◽  
pp. 253-262 ◽  
Author(s):  
Alexei V Tulin ◽  
Galina L Kogan ◽  
Dominik Filipp ◽  
Maria D Balakireva ◽  
Vladimir A Gvozdev

The 30-kb cluster comprising close to 20 copies of tandemly repeated Stellate genes was localized in the distal heterochromatin of the X chromosome. Of 10 sequenced genes, nine contain undamaged open reading frames with extensive similarity to protein kinase CK2 β-subunit; one gene is interrupted by an insertion. The heterochromatic array of Stellate repeats is divided into three regions by a 4.5-kb DNA segment of unknown origin and a retrotransposon insertion: the A region (∼14 Stellate genes), the adjacent B region (approximately three Stellate genes), and the C region (about four Stellate genes). The sequencing of Stellate copies located along the discontinuous cluster revealed a complex pattern of diversification. The lowest level of divergence was detected in nearby Stellate repeats. The marginal copies of the A region, truncated or interrupted by an insertion, escaped homogenization and demonstrated high levels of divergence. Comparison of copies in the B and C regions, which are separated by a retrotransposon insertion, revealed a high level of diversification. These observations suggest that homogenization takes place in the Stellate cluster, but that inserted sequences may impede this process.


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