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Biomedical subjects

E Karsenti

Publications and source records attributed to E Karsenti.

97 records · Page 6Linked to original sources

Receptor mobility and its cooperative restriction by ligands.

The functional significance of membrane fluidity and receptor mobility in lymphoid cells has been studied in the recent literature. Although far from clarified, the role of membrane fluidity in achieving control over cell activity is probably important; it allows cooperative interactions over long distances. Here, the emphasis is put on the phenomenon of restriction of receptor mobility by ligands such as Concanavalin A, a phenomenon discovered in recent years using morphological techniques. We discuss in some detail our own approach for studying this phenomenon. This consists of quantitatively measuring the active sites on cell-bound lectin molecules by subsequent fixation of horse-radish peroxidase. This study has shown a cooperative binding of Concanavalin A to cells which corresponds to a modification of the membrane, leading to the recruitment of new receptors. The existence of a post-binding event, that we have called micro-redistribution, has been shown at 4 degrees C, through the use of peroxidase binding to cell-bound lectin. A cooperative restriction of receptor microredistribution is observed when the cooperative recruitment of receptors induced by increasing concentrations of Concanavalin A occurs. Both phenomena were shown to be modulated by drugs such as colchicine and cytochalasin B. The characteristics of this modulation suggest that density and distribution of receptors are dependent upon the state of a multimeric submembrane structure which is still functional at 4 degrees C.

Lectins↗

A model for the proposed roles of different microtubule-based motor proteins in establishing spindle bipolarity.

BACKGROUND: In eukaryotes, assembly of the mitotic spindle requires the interaction of chromosomes with microtubules. During this process, several motor proteins that move along microtubules promote formation of a bipolar microtubule array, but the precise mechanism is unclear. In order to examine the roles of different motor proteins in building a bipolar spindle, we have used a simplified system in which spindles assemble around beads coated with plasmid DNA and incubated in extracts from Xenopus eggs. Using this system, we can study spindle assembly in the absence of paired cues, such as centrosomes and kinetochores, whose microtubule-organizing properties might mask the action of motor proteins. RESULTS: We blocked the function of individual motor proteins in the Xenopus extracts using specific antibodies. Inhibition of Xenopus kinesin-like protein 1 (Xklp1) led either to the dissociation of chromatin beads from microtubule arrays, or to collapsed microtubule bundles on beads. Inhibition of Eg5 resulted in monopolar microtubule arrays emanating from chromatin beads. Addition of antibodies against dynein inhibited the focusing of microtubule ends into spindle poles in a dose-dependent manner. Inhibition of Xenopus carboxy-terminal kinesin 2 (XCTK2) affected both pole formation and spindle stability. Co-inhibition of XCTK2 and dynein dramatically increased the severity of spindle pole defects. Inhibition of Xklp2 caused only minor spindle pole defects. CONCLUSIONS: Multiple microtubule-based motor activities are required for the bipolar organization of microtubules around chromatin beads, and we propose a model for the roles of the individual motor proteins in this process.

Animals↗

Release of mature starfish oocytes from interphase arrest by microinjection of human centrosomes.

Mature oocytes (unfertilized eggs) are arrested at definite cell-cycle stages which vary from species to species. In frogs and mammals, the oocytes are arrested at the second metaphase of meiosis whereas in echinoderms they are blocked later, at the pronucleus stage. What causes the maturing oocytes to stop at some point in the cell cycle is not entirely clear. In frogs, the metaphase arrest seems to be maintained by a cytostatic factor. In echinoderms, which stop at interphase, no such a factor has so far been found. The fertilization process, beyond the introduction of paternal chromosomes, releases the oocyte from cell-cycle arrest and provides a functional centrosome to replace the endogenous centrosome which is apparently lost during oogenesis in most species. Several lines of evidence suggest that release from cell-cycle arrest is mediated by a Ca2+ burst which is associated with fertilization, and it is known that the functional centrosome provided by the sperm is necessary for mitotic spindle formation and cleavages. We report here that microinjection of purified human centrosomes into mature starfish oocytes is sufficient to release them from arrest at interphase and to support many cleavages leading to the occasional formation of normal embryos. In this species centrosome induced re-entry into the cell cycle does not require a transient calcium burst nor does it require intact microtubules.

Animals↗

[Anti-actin and anti-tubulin antibodies in the serum of non-immunized animals].

The preparation and the purification of anti-action and anti-tubulin antibodies is reported. The occurrence of low amount of IgG anti-tubulin was also found in the sera of non-immunized animals from various species. The staining of the cellular network of microtubules as well as of tubulin paracrystals was observed with both induced and natural antibodies.

Actins↗