Integrins: regulators of embryogenesis.

Integrins are heterodimeric transmembrane glycoproteins involved in cell-cell and cell-extracellular matrix adhesion. They also participate in cytoskeletal rearrangements, co-regulation of growth factor activities and activation of signal transductions. This review describes experimental approaches that have given new insights into the integrin functions during embryogenesis. Using anti-functional antibodies, peptide inhibitors of integrin-ligand interactions and genetic ablation of integrins results, this review will show that integrins are key molecules during early development of both invertebrates and vertebrates.

Polar organization of gamma-tubulin in acentriolar mitotic spindles of Drosophila melanogaster cells.

The spindle pole localization of gamma-tubulin was compared in wild type and acentriolar cultured Drosophila cells using polyclonal antibodies specifically raised against the carboxy terminal amino acid sequence of Drosophila gamma-tubulin-1 (-KSEDSRSVTSAGS). During interphase, gamma-tubulin was present in the centrosome of wild type cells and accumulated around this organelle in a cell cycle dependent manner. In contrast, no such structure was observed in acentriolar cells. Wild type mitoses were homogeneously composed of biconical spindles, with two centrosome-associated gamma-tubulin spots at the poles. The mitotic apparatuses observed in the acentriolar cells were heterogeneous; multipolar mitoses, bipolar mitoses with a barrel-shaped spindle and bipolar mitoses with biconical spindles were observed. In acentriolar cells, gamma-tubulin accumulation at mitotic poles was dependent on spindle microtubule integrity. Most acentriolar spindles presented a dispersed gamma-tubulin labeling at the poles. Only well polarized and biconical acentriolar spindles showed a strong gamma-tubulin polar spot. Finally, acentriolar mitotic poles were not organized around true centrosomes. In contrast to wild type cells, in acentriolar cells the Bx63 centrosome-associated antigen was absent and the gamma-tubulin containing material dispersed readily following microtubule disassembly. These observations confirm that gamma-tubulin plays an essential role in the nucleation of microtubules even in the absence of mitotic polar organelles. In addition the data suggest that the mechanisms involved in the bipolarization of wild type and acentriolar mitoses are different, and that centrioles play a role in the spatial organization of the nucleating material containing gamma-tubulin.

Effect of hydrogen peroxide on cytoskeletal proteins of Drosophila cells: comparison with heat shock and other stresses.

Hydrogen peroxide, which was shown to trigger the heat-shock response by activating the immediate binding of the heat-shock factor to DNA heat shock regulatory elements in the promoter of heat-shock genes of Drosophila cells, has also been reported to enhance the synthesis of actin. We show here that very short and transient H2O2 treatments, from 1 s to 2 min, are sufficient to induce an increase of actin synthesis. This increase becomes apparent 2 to 3 h after the short H2O2 treatment. It is inhibited if actinomycin D is present during the short H2O2 treatment. An increase of actin synthesis was also observed during the recovery period after two other stresses: reoxygenation after anoxia and ethanol treatment. The synthesis of two cytoskeletal proteins, tubulin and a 46-kDa insoluble protein of the intermediate filament fraction, was also slightly increased by H2O2 in Drosophila cells, but this increase was not actinomycin D-dependent. H2O2 does not provoke the translocation of the 46-kDa protein to the nuclear fraction as does heat shock. The very rapid stimulation of actin synthesis by H2O2 and the involvement of cytoskeletal elements in many stress situations suggest that actin may play a key role in the response to external stimuli.

Cyclin B is associated with centrosomes in Drosophila mitotic cells.

We have studied by way of confocal laser scanning microscopy the subcellular localization of cyclin B in Drosophila-cultured cells and report here evidence that a part of the cyclin B cell pool is closely associated with the centrosome. This cyclin B centrosomal signal is strong in prophase and metaphase but disappears during anaphase. Moreover, the signal is absent in the acentriolar Drosophila cell line 1182-4. These results put forward additional arguments suggesting that the centrosome plays an important role in the control of the cell cycle.

Overall study of the in vitro plasma clotting system in an invertebrate, Liocarcinus puber (Crustacea Decapoda): considerations on the structure of the Crustacea plasma fibrinogen in relation to evolution.

An overall study of the in vitro plasma coagulation system in the crab Liocarcinus puber has been carried out using various analytical methods, namely thromboelastography, spectrophotometrical examination, and a new one based on changes of the mechanical impedance of the developing clot. From the results reported here the clotting pattern in this species appears surprisingly complex for an invertebrate and unexpectedly closer to that of the vertebrates. Indirect evidences suggest that the fibrinogen polypeptide chains in this species and very likely in the other crustacean, are very different from those of the vertebrates. This would imply that crustacean and vertebrate fibrinogen would have diverged from one another in a far remote past, far beyond the individualization of the vertebrate alpha chain, that is, over 1.5 million years ago.

Cross induced coagulations between human and crustacean clotting factors. Considerations on the clotting processes.

1. Cross induced coagulations show that human factor XIII and crustacean coagulin are to some extent functionally equivalent and may be substituted for each other. 2. In crustacea, fibrinogen and coagulin appear as in situ activated products since they are both able to react with non-activated human clotting factors. 3. The coagulin catalyzed transamidation which stabilizes the clot and renders it insoluble in 1-5% monochloroacetic acid solutions seems to be the basic reaction of the clotting process in the animals in which coagulation occurs. 4. The possibility of a two step clotting in crustacea is discussed.

[Coagulation of hemolymph in Crustacea. Electrophoretic demonstration of a plasmatic component in the course of hemostasis in a group A crustacean (Cancer pagurus)]. / Coagulation de l'hémolymphe chez les crustacés. Mise en évidence par électrophorèse d'un composant plasmatique intervenant dans l'hémostase chez un crustacé du groupe A (Cancer pagurus).

Coagulation assays in vitro of plasmatic extract from Cancer pagurus reveal the existence of a plasmatic coagulation in this species which was considered as yet as lacking a such process. Electrophoretical analysis of this extract before and after coagulation shows the presence of a plasmatic component probably involved in the formation of the clot.

[Coagulation of the hemolymph in Macropipus puber (L)., Crustacea, Decapoda. Electrophoretic demonstration of a plasma component intervening in hemostasis]. / Coagulation de l'hémolymphe chez Macropipus puber (L.). Crustacé décapode. Mise en évidence par électrophorèse d'un composant plasmatique intervenant dans l'hémostase.

Electrophoretical and immuno-electrophoretical analysis of plasma, serum and some plasmatic extracts of Macropipus puber (L). have evidenced the presence in the plasma of a component absent in the serum. This component which disappears from the plasmatic extracts after their coagulation must therefore plays a role in the clotting processes.

[Hemolymph coagulation in Macropipus puber (L.) Crustacea Decapoda. Respective roles of hemolymph and plasma]. / Coagulation de l'hemolymphe chez Macropipus puber (L.) Crustacé Décapode. Mise en évidence des rôles respectifs du plasma et des organites cellulaires dans les mécanismes de la coagulation

The process of coagulation of the hemolymph in Macropipus puber (L.) Crustacea Decapod depends on one or several plasmatic factors and on a thermo-labile cellular factor which does not seem to be specific. The existence of these factors has been demonstrated.