Biomedical subjects

M G Riparbelli

Publications and source records attributed to M G Riparbelli.

33 records · Page 2Linked to original sources

PubMed1994

The proliferating cell marker monoclonal antibody Ki-67 recognizes specific antigens associated with the nuclear envelope of the early Drosophila embryo.

Immunofluorescence and immunoelectron microscopy indicated that the antibody raised against the nuclear antigen Ki-67 of mammalian cells recognized antigenic determinants of early Drosophila embryos, localized on the outside of the nuclear envelope. Hence, the nuclear envelope of Drosophila appears to share a similar epitope with the chromosome scaffold of mitotic mammalian cells. With the progression of mitosis the antigen persisted around the mitotic spindle region and was also found in the pole regions at metaphase and anaphase. The antibody also stained the equatorial regions of the spindles from anaphase to late telophase. The antibody may therefore be used as a biochemical marker of the nuclear envelope for studying nuclear membrane biogenesis and behavior during the mitotic divisions of the Drosophila embryo.

Anaphase↗

PubMed1994

The distribution of cytoplasmic bacteria in the early Drosophila embryo is mediated by astral microtubules.

Maternally inherited cytoplasmic bacteria have occasionally been observed in embryos and adults of different strains of several Drosophila species. While there is a considerable body of data on the relationship between bacteria and embryo viability, little is known about the behavior of these bacteria during the early development of Drosophila. In eggs laid by infected Drosophila melanogaster females we showed that cytoplasmic bacteria were initially concentrated in a thin cortical layer and scattered in the yolk region. During the following syncytial blastoderm mitoses the bacteria mainly accumulated towards the poles of the mitotic spindles, suggesting that astral microtubules play a role in localizing bacteria. This is supported by the observation that treatment of the infected embryos with the microtubule-disrupting drug colchicine led to the partial dissociation of the bacteria from the spindle poles, whereas cytochalasin treatment left almost all the bacterial clusters intact. Moreover, bacteria were not found near the polar bodies and yolk nuclei, which were without astral microtubules. In mitosis-defective embryos, with centrosomes dissociated from the nuclei, the bacteria were concentrated in association with the isolated astral microtubules, and in cold-treated embryos, in which microtubules regrew from isolated centrosomes after recovering, the bacteria clustered around the newly formed asters. These observations, also supported by electron microscope analysis, indicate a close relationship between cytoplasmic bacteria and astral microtubules, and suggest that the latter were able to build discrete cytoplasmic domains ensuring the proper distribution of cytoplasmic components during the blastoderm mitoses, despite the lack of cell membranes.

Animals↗

PubMed1993

Spatial organization of microtubules and microfilaments in larval and adult salivary glands of Drosophila melanogaster.

We examined the distribution of microtubules and microfilaments by conventional fluorescence microscopy and laser scanning confocal microscopy in larval and adult salivary glands of Drosophila melanogaster. The cells of the larval salivary gland epithelium were characterized by the same spatial distribution of microfilaments, whereas microfilament localization was more complex in adult salivary glands, showing some regional differentiation. Microtubules distributed throughout the cell cytoplasm of the larval salivary glands, whereas in adult glands they were mostly observed in the basal or apical cytoplasm of the cells. These observations were related to the secretory process and the mechanism of saliva discharge.

Actin Cytoskeleton↗

PubMed1993

The cortical actin cytoskeleton in a Dipteran embryo: analysis of the spatial reorganization of F-actin aggregates during the early nuclear division cycles.

Rhodamine phalloidin-staining was used to study the organization of the cortical actin cytoskeleton of the early Ceratitis capitata embryo. The dynamics of the actin aggregates and their changes in distribution during the formation of the syncytial blastoderm, were followed in detail. It was found that these aggregates formed a shell-like cluster around the interphase nuclei, and concentrated toward the poles of the mitotic apparatus when the nuclei divided. Laser scanning confocal microscopy revealed that aggregates not clustered at the poles of the mitotic apparatus were closely associated with fine fibers of a dense cytoplasmic network of actin filaments.

Actins↗

PubMed1992

Surface cap modifications in cold-treated Drosophila melanogaster embryos.

When early Drosophila embryos were allowed to develop at 0 degree C, several abnormalities in the surface cap organization were observed. Scanning electron microscopy showed that exposure to cold mainly lead to the deformation of the cortical caps and to their partial fusion with adjacent caps. The process of cellularization was presumably affected and large uncellularized areas were observed. Rhodamine-phalloidin staining showed that cap deformation was closely related to the altered microfilament distribution, which was presumably responsible for the failure of large syncytial areas to cellularize. During the process of cellularization, F-actin localization did not depend on the microtubules forming the baskets around the elongating nuclei, but was related to the subpopulation of microtubules radiating from the centrosomes toward the plasma membrane. Only these microtubules seemed to be affected by cold treatment.

Actin Cytoskeleton↗

PubMed1992

Involvement of microtubules and microfilaments in centrosome dynamics during the syncytial mitoses of the early Drosophila embryo.

To examine the role of microfilaments and microtubules in centrosome dynamics we exposed Drosophila embryos to culture medium containing cytochalasin B and to low temperature. The results show that the splitting of the centrosomal material does not occur when the embryos are treated with cytochalasin before centrosome duplication at late telophase. The fragmentation of the centrosomal material, caused by cold exposure, is also prevented by cytochalasin incubation. These results indicate that both microtubules and microfilaments may be involved in determining centrosome shape during the syncytial mitoses which lead to the formation of the blastoderm in early Drosophila embryos.

Actin Cytoskeleton↗

PubMed1992

Cytochalasin induces spindle fusion in the syncytial blastoderm of the early Drosophila embryo.

Microfilament integrity is needed to maintain the regular arrangement of the spindle microtubules and to guarantee the normal progression of the last syncytial mitoses in Drosophila embryo. To investigate when and how microfilaments participate in this process, we incubated permeabilized embryos with the inhibitor of actin polymerization, cytochalasin B, at different times during the nuclear cycle. Our results suggest that the correct microfilament distribution is only required for the appropriate segregation of nuclei during the 11th, 12th and 13th syncytial mitoses rather than during the 10th mitosis when the spindles are too far apart to interact. When cytochalasin B treatment was performed during the last syncytial mitoses many spindles fuse among them and the regular mitotic progression is perturbed.

Actin Cytoskeleton↗

PubMed1992

Distribution of a nuclear envelope antigen during the syncytial mitoses of the early Drosophila embryo as revealed by laser scanning confocal microscopy.

The changing distribution of a nuclear envelope antigen recognized by a monoclonal antibody raised against human fibroblast vimentin during the syncytial mitoses of the Drosophila embryo has been studied with a confocal laser scanning microscope. The antigen appears very early as irregular aggregates in the peripheral cytoplasm of the preblastoderm embryo. As the first nuclei reach the periplasm the antigen is localized on the nuclear envelope and the cytoplasmic staining decreases. In addition to the perinuclear labeling we observed intense midzone and polar staining during the mitotic cycle. A possible relationship between polar localization of the antigen and centrosome position is discussed.

Animals↗

PubMed1991

A monoclonal antibody recognizing a common antigen on Drosophila embryos and human fibroblasts.

We used a monoclonal antibody specific for vimentin from human fibroblasts to stain whole mounts of Drosophila embryos. In immunofluorescence observations this antibody cross-reacts with an antigenic determinant localized throughout mitosis at the nuclear boundary. Double fluorescence observations with the Rb188 antibody that specifically recognizes a centrosomal protein of the Drosophila embryo [Whitfield et al., 1988] showed that the anti-vimentin antibody cross-reacts with an antigen localized in the centrosomal region.

Animals↗

PubMed1991

Microfilament distribution in cold-treated Drosophila embryos.

Cold treatment of Drosophila embryos is observed to result in general alteration of microfilament distribution leading to deformation of the surface caps and to perturbation of the process of cleavage furrow extension. After exposure to low temperature the cortical actin caps underwent several morphological changes, despite the arrested nuclear cycle. These observations are discussed in relation to centrosome behavior during the cell cycle.

Actin Cytoskeleton↗

PubMed1990

Centriole and centrosome cycle in the early Drosophila embryo.

Centriole and centrosome cycles were examined by indirect immunofluorescence and electron microscopy techniques in the early Drosophila embryo. The centrosomes, which are already divided at interphase, appear as compact spheres during prophase and metaphase, expand and flatten from anaphase to telophase and split into two units in late telophase. Centriole separation starts in late metaphase, becomes evident in anaphase and increases during telophase. Procentrioles appear during the following interphase.

Animals↗

PubMed1989

Diazepam induces abnormal mitosis in the early Drosophila embryo.

Drosophila embryos, because of their high proportion of dividing nuclei, offer many advantages for the study of the mitotic cycle. In the present study we combined immunofluorescence with interference contrast techniques to follow centrosome and spindle behavior in embryos exposed to diazepam during the first stages of development. Exposure to 100 micrograms/ml of diazepam produced polyploid and aneuploid figures resulting from the unusual fusion of one or more adjacent spindles. Diazepam also causes the inhibition of centrosome shifting and induces the formation of monopolar spindles during the metaphase-anaphase transition.

Animals↗

PubMedDate not supplied

Mechanisms of microsporidial cell division: ultrastructural study on Encephalitozoon hellem.

The mitotic process in microsporidian Encephalitozoon hellem, a known human pathogen, has been studied with the aim of elucidating some ultrastructural aspects of its nuclear division. The presence of a nuclear spindle, of "electrondense spindle plaques" associated with the nuclear envelope and of cytoplasmic double walled vesicles are reported. We suggest that these "electrondense spindle plaques" serve as foci for intranuclear and cytoplasmic microtubule arrangements, similar to the microtubule organizing centers within the centrosomes of animal cells. The extent to which the microsporidial division process is comparable with that of more familiar eukaryotes such as yeast cells is discussed.

AIDS-Related Opportunistic Infections↗

PubMedDate not supplied

Evidence of actin in the cytoskeleton of microsporidia.

Using transmission electron microscopy, immuno-electron microscopy, and biochemical techniques such as 2-D electrophoresis and immunoblotting, actin was found in all biological stages of the microsporidia Encephalitozoon hellem and Encephalitozoon cuniculi.

Actins↗

PubMedDate not supplied

Centrosome inheritance in insects: fertilization and parthenogenesis.

Centrosome biogenesis is unclear, although much structural and biochemical research has been performed in several experimental systems. An alternative model to study the assembly of functional centrosomes could be the process of zygotic centrosome formation at the beginning of embryonic development. Although it seems obvious that the sperm cell provides the centrosome at fertilization, some pieces of evidence are not in line with this point of view and give controversial results. Such an analysis could provide useful information if applied to a large variety of organisms. Since insects are a highly diverse group of organisms they provide a variety of models in which to study the process of centrosome reconstitution during fertilization. Moreover, many insect species reproduce by parthenogenesis, a special mode of reproduction in which embryonic development occurs without male contribution. Studies of unfertilized parthenogenetic eggs may therefore teach us much about the process of centrosome assembly in the absence of preexisting centrioles.

Animals↗