scholarly journals Specific interactions of chromatin with the nuclear envelope: positional determination within the nucleus in Drosophila melanogaster.

1996 ◽  
Vol 7 (5) ◽  
pp. 825-842 ◽  
Author(s):  
W F Marshall ◽  
A F Dernburg ◽  
B Harmon ◽  
D A Agard ◽  
J W Sedat
Keyword(s):  
Drosophila Melanogaster ◽  
Nuclear Envelope ◽  
Three Dimensional ◽  
Specific Interactions ◽  
Early Embryos ◽  
Chromatin Proteins ◽  
Discrete Site ◽  
Large Stretch ◽  
Drosophila Embryos

Specific interactions of chromatin with the nuclear envelope (NE) in early embryos of Drosophila melanogaster have been mapped and analyzed. Using fluorescence in situ hybridization, the three-dimensional positions of 42 DNA probes, primarily to chromosome 2L, have been mapped in nuclei of intact Drosophila embryos, revealing five euchromatic and two heterochromatic regions associated with the NE. These results predict that there are approximately 15 NE contacts per chromosome arm, which delimit large chromatin loops of approximately 1-2 Mb. These NE association sites do not strictly correlate with scaffold-attachment regions, heterochromatin, or binding sites of known chromatin proteins. Pairs of neighboring probes surrounding one NE association site were used to delimit the NE association site more precisely, suggesting that peripheral localization of a large stretch of chromatin is likely to result from NE association at a single discrete site. These NE interactions are not established until after telophase, by which time the nuclear envelope has reassembled around the chromosomes, and they are thus unlikely to be involved in binding of NE vesicles to chromosomes following mitosis. Analysis of positions of these probes also reveals that the interphase nucleus is strongly polarized in a Rabl configuration which, together with specific targeting to the NE or to the nuclear interior, results in each locus occupying a highly determined position within the nucleus.

Detection and mapping of interactions between chromatin and the nuclear envelope in drosophila embryos

1995 ◽  
Vol 53 ◽  
pp. 764-765
Author(s):  
W.F. Marshall ◽  
A.F. Dernburg ◽  
B. Harmon ◽  
J.W. Sedat
Keyword(s):  
Nuclear Envelope ◽  
Chromatin Condensation ◽  
Three Dimensional ◽  
Physiological Role ◽  
Nuclear Surface ◽  
Intact Cells ◽  
Molecular Determinants ◽  
Surface Harmonic ◽  
Drosophila Embryos

Interactions between chromatin and nuclear envelope (NE) have been implicated in chromatin condensation, gene regulation, nuclear reassembly, and organization of chromosomes within the nucleus. To further investigate the physiological role played by such interactions, it will be necessary to determine which loci specifically interact with the nuclear envelope. This will not only facilitate identification of the molecular determinants of this interaction, but will also allow manipulation of the pattern of chromatin-NE interactions to probe possible functions. We have developed a microscopic approach to detect and map chromatin-NE interactions inside intact cells.Fluorescence in situ hybridization (FISH) is used to localize specific chromosomal regions within the nucleus of Drosophila embryos and anti-lamin immunofluorescence is used to detect the nuclear envelope. Widefield deconvolution microscopy is then used to obtain a three-dimensional image of the sample (Fig. 1). The nuclear surface is represented by a surface-harmonic expansion (Fig 2). A statistical test for association of the FISH spot with the surface is then performed.

Three-dimensional reconstruction of the chromatin bodies in the nuclei of mature erythrocytes from the newt Triturus cristatus: the number of nuclear envelope-attachment sites

1979 ◽  
Vol 35 (1) ◽  
pp. 59-66
Author(s):  
A.B. Murray ◽  
H.G. Davies
Keyword(s):  
Nuclear Envelope ◽  
Three Dimensional ◽  
Interphase Nuclei ◽  
Interphase Chromosome ◽  
Triturus Cristatus ◽  
3 Dimensional ◽  
Attachment Sites ◽  
Chromatin Body ◽  
Dimensional Reconstruction ◽  
Discrete Site

The arrangement of the chromatin bodies in the interphase nuclei of 6 erythrocytes has been investigated by means of 3-dimensional reconstruction from electron micrographs of serial sections. When the borders of chromatin bodies are marked on the surface of each model, discrete areas of chromatin in contact with the nuclear envelope are revealed. The number of these areas in approximately equal to the number of chromosomes in the diploid set. The data suggest that each chromatin body corresponds to a condensed interphase chromosome and that each chromosome is attached to one discrete site on the nuclear envelope. The data are insufficient to show whether or not the condensed chromosomes are arranged in any orderly pattern in these nuclei.

DNA from Drosophila melanogaster β-heterochromatin binds specifically to nuclear lamins in vitro and the nuclear envelope in situ

Gene ◽  
1996 ◽  
Vol 171 (2) ◽  
pp. 171-176 ◽  
Author(s):  
Ella A. Baricheva ◽  
Miguel Berrios ◽  
Sergei S. Bogachev ◽  
Igor V. Borisevich ◽  
Eugenia R. Lapik ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Nuclear Envelope ◽  
Nuclear Lamins

scholarly journals Three-dimensional organization of Drosophila melanogaster interphase nuclei. II. Chromosome spatial organization and gene regulation.

1987 ◽  
Vol 104 (6) ◽  
pp. 1471-1483 ◽  
Author(s):  
M Hochstrasser ◽  
J W Sedat
Keyword(s):  
Gene Regulation ◽  
Drosophila Melanogaster ◽  
Nuclear Envelope ◽  
Spatial Organization ◽  
Polytene Chromosomes ◽  
Three Dimensional ◽  
Nucleolar Organizer ◽  
Cell Types ◽  
Functional Changes ◽  
Specific Subset

In the preceding article we compared the general organization of polytene chromosomes in four different Drosophila melanogaster cell types. Here we describe experiments aimed at testing for a potential role of three-dimensional chromosome folding and positioning in modulating gene expression and examining specific chromosome interactions with different nuclear structures. By charting the configurations of salivary gland chromosomes as the cells undergo functional changes, it is shown that loci are not repositioned within the nucleus when the pattern of transcription changes. Heterologous loci show no evidence of specific physical interactions with one another in any of the cell types. However, a specific subset of chromosomal loci is attached to the nuclear envelope, and this subset is extremely similar in at least two tissues. In contrast, no specific interactions between any locus and the nucleolus are found, but the base of the X chromosome, containing the nucleolar organizer, is closely linked to this organelle. These results are used to evaluate models of gene regulation that involve the specific intranuclear positioning of gene sequences. Finally, data are presented on an unusual class of nuclear envelope structures, filled with large, electron-dense particles, that are usually associated with chromosomes.

scholarly journals In situ Nuclear Matrix preparation in Drosophila melanogaster and its use in studying the components of nuclear architecture

2021 ◽  
Author(s):  
Rashmi U Pathak ◽  
Rahul Sureka ◽  
Ashish Bihani ◽  
Parul Varma ◽  
Rakesh K Mishra
Keyword(s):  
Drosophila Melanogaster ◽  
Nuclear Matrix ◽  
Cell Biology ◽  
Three Dimensional ◽  
Nuclear Architecture ◽  
Isolated Nuclei ◽  
Intact Embryo ◽  
Matrix Preparation ◽  
Transgenic Flies

The study of Nuclear Matrix (NuMat) over the last 40 years has been limited to either isolated nuclei from tissues or cells grown in culture. Here, we provide a protocol for NuMat preparation in intact Drosophila melanogaster embryos and its use in dissecting the components of nuclear architecture. The protocol does not require isolation of nuclei and therefore maintains the three-dimensional milieu of an intact embryo, which is biologically more relevant compared to cells in culture. One of the advantages of this protocol is that only a small number of embryos are required. The protocol can be extended to larval tissues like salivary glands and imaginal discs with little modification. Taken together, it becomes possible to carry out such studies in parallel to genetic experiments using mutant and transgenic flies. This protocol, therefore, opens the powerful field of fly genetics to cell biology in the study of nuclear architecture.

scholarly journals Temporal and spatial coordination of chromosome movement, spindle formation, and nuclear envelope breakdown during prometaphase in Drosophila melanogaster embryos.

1990 ◽  
Vol 111 (6) ◽  
pp. 2815-2828 ◽  
Author(s):  
Y Hiraoka ◽  
D A Agard ◽  
J W Sedat
Keyword(s):  
Drosophila Melanogaster ◽  
Nuclear Envelope ◽  
Temporal Dynamics ◽  
Three Dimensional ◽  
Metaphase Plate ◽  
Time Lapse ◽  
Three Dimensions ◽  
Nuclear Envelope Breakdown ◽  
High Resolution Analysis ◽  
Nuclear Structures

The spatial and temporal dynamics of diploid chromosome organization, microtubule arrangement, and the state of the nuclear envelope have been analyzed in syncytial blastoderm embryos of Drosophila melanogaster during the transition from prophase to metaphase, by three-dimensional optical sectioning microscopy. Time-lapse, three-dimensional data recorded in living embryos revealed that congression of chromosomes (the process whereby chromosomes move to form the metaphase plate) at prometaphase occurs as a wave, starting at the top of the nucleus near the embryo surface and proceeding through the nucleus to the bottom. The time-lapse analysis was augmented by a high-resolution analysis of fixed embryos where it was possible to unambiguously trace the three-dimensional paths of individual chromosomes. In prophase, the centromeres were found to be clustered at the top of the nucleus while the telomeres were situated at the bottom of the nucleus or towards the embryo interior. This polarized centromere-telomere orientation, perpendicular to the embryo surface, was preserved during the process of prometaphase chromosome congression. Correspondingly, breakdown of the nuclear envelope started at the top of the nucleus with the mitotic spindle being formed at the positions of the partial breakdown of the nuclear envelope. Our observation provide an example in which nuclear structures are spatially organized and their functions are locally and coordinately controlled in three dimensions.

The Localization of β-Glucuronidase in the Early Chick Embryo

Development ◽  
1958 ◽  
Vol 6 (1) ◽  
pp. 52-56
Author(s):  
F. Billett ◽  
Leela Mulherkar
Keyword(s):  
Drosophila Melanogaster ◽  
Chick Embryo ◽  
Embryonic Development ◽  
Xenopus Laevis ◽  
Mouse Liver ◽  
Histochemical Method ◽  
Toxic Compounds ◽  
Early Chick Embryo ◽  
Early Embryos ◽  
Drosophila Embryos

In vertebrate tissues there appears to be a connexion between β-glucuronidase and the proliferation of cells. This connexion was first noticed by Levvy, Kerr, & Campbell (1948) when they were investigating the effect of toxic compounds on mouse-liver glucuronidase. The early stages of embryonic development are characterized by rapid proliferations of cells and it is, therefore, of some interest to study the localization of β-glucuronidase during these stages. Small amounts of β-glucuronidase can be detected in the early embryos of Xenopus laevis (Billett, 1956) and in those of Drosophila melanogaster (Billett & Counce, unpublished). In these embryos no marked increase in the enzyme can be associated with the proliferation of cells. The large amount of yolk in the Xenopus and Drosophila embryos was a complicating factor in the above experiments. It was not possible to localize the enzyme in these embryos with a histochemical method.

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