Relations entre la prolifération et la différenciation cellulaires dans l'intestin embryonnaire et larvaire dePleurodeles waltlii Michah

Résumé Les chalones 1 et 2, extraites de l'intestin de pleurodèle adulte, inhibent en phase G₁ et G₂ respectivement les cycles mitotiques de l'épithélium intestinal embryonnaire. Les effets de ces chalones sur la prolifération et la différenciation cellulaires dans ce tissu ont été étudiés en fonction de la dose injectée, du stade de développement et de la durée du traitement. L'inhibition provoquée par la chalone 2 est proportionnelle à la dose injectée entre deux seuils de concentration. Le quart environ des cellules intestinales en activité mitotique est insensible à la chalone 2 même à la suite d'injections répétées de l'inhibiteur. Seules les cellules intestinales des embryons âgés (stade 34) sont sensibles à cette chalone et répondent par un allongement de la phase G₂ qui, malgré des injections répétées de l'inhibiteur, n'excède pas une vingtaine d'heures. La sensibilité des cellules de l'épithélium intestinal à la chalone 1 se manifeste à la fin du développement (stade 33), comme dans le cas de la chalone 2. A l'égard de la chalone 1, la population cellulaire en activité mitotique dans l'intestin embryonnaire apparaît hétérogène et comprend: 50% de cellules aptes à être inhibées par des doses faibles de chalone 1; 25% de celludes aptes à n'être inhibées que par des doses de chalone 1 cent fois plus élevées et 25% environ de cellules insensibles à cet inhibiteur. Les injections répétées de chalone 1 bloquent définitivement en phase G₁ la moitié environ des cellules en activité mitotique dans l'épithélium intestinal indifférencié au stade 34; en outre, elles accélèrent la consommation du vitellus, favorisent la différenciation des cellules à mucus et diminuent le nombre des cellules constituant les nids sous-épithéliaux qui apparaissent au stade 36 et représentent le compartiment générateur de l'épithélium intestinal. Les réultats obtenus permettent de proposer un modèle de cinétique de la prolifération cellulaire au cours de la différenciation de l'épithélium intestinal du pleurodèle; de plus, ils conduisent à l'hypothèse que le nombre de divisions subies par une cellule embryonnaire et le taux de chalone dans le tissu auquel elle appartient, sont les deux signaux complémentaires qui déclenchent le blocage du cycle mitotique et l'achèvement de la différenciation dans cette cellule.

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Relationships between the cell proliferation and the differentiation in the embryonic and larval intestine ofpleurodeles waltlii michah. II. Effects of intestinal chalones extracted from the intestine of the adult newt

Summary The intestinal chalones 1 and 2, extracted from the intestine of the adult newt, are known to inhibit the G₁ and G₂ phases of the cell cycle in the embryonic intestine. The effects of these intestinal chalones on the proliferation and differentiation of intestinal cells of newt embryos were studied with special attention to the dose-response relationship, the embryonic stage and the duration of treatment. The chalone 2 triggered a linear, dose-dependent inhibition between two concentration thresholds; nevertheless about 25% of the cycling cells were not inhibited either by the highest doses injected or by repeated injections. Sensitivity to chalone 2 appeared in the intestinal epithelium at the end of embryonic development (stage 34) but the cells were not delayed in the G₂ phase for more than about 20 h in spite of repeated injections. It was inferred from the doseresponse curve of the mitotic inhibition by chalone 1, that the intestinal cell population was heterogeneous: about 50% of the cycling cells were inhibited by low concentrations of chalone 1; an additional proportion of about 25% of cycling cells was inhibited by 100 x more concentrated chalone 1 and the remaining 25% was insensitive to the inhibitor. Repeated injections of chalone 1 blocked about 50% of the cycling cells definitively in the G₁ phase, speeded up digestion of yolk platelets, promoted the differentiation of goblet cells and depressed the number of stem cells in the proliferative compartment located beneath the epithelium. A kinetics model of cell proliferation and cell differentiation in the intestinal cell lineages was elaborated and it was suggested that the arrest of mitotic activity and the completion of differentiation in an embryonic cell depends on two incoming signals: one is intracellular and appears when the required number of cell cycles has occured in the cell lineage, leading to a committed stem cell sufficiently differentiated to synthesise chalone and to respond to chalone; the other signal is extracellular and appears when the chalone concentration is high enough: i.e. when the required number of cells is obtained in this tissue.

Ce travail a bénéficié de l'aide du CNRS (ATP N^o A655 1799) et de l'INSERM (AT N^o 74142036)     

Biomimetic synthesis of the IDO inhibitors exiguamine A and B

Biomimetic synthesis is an attempt to assemble natural products along biosynthetic lines without recourse to the full enzymatic machinery of nature. We exemplify this with a total synthesis of exiguamine A and the newly isolated natural product exiguamine B. The most noteworthy feature of this work is an oxidative endgame drawing from the complex chemistry of catecholamines, which allows for ready access to a new class of nanomolar indoleamine-2,3-dioxygenase inhibitors.     

Evolving concepts in cancer therapy through targeting sphingolipid metabolism

Traditional methods of cancer treatment are limited in their efficacy due to both inherent and acquired factors. Many different studies have shown that the generation of ceramide in response to cytotoxic therapy is generally an important step leading to cell death. Cancer cells employ different methods to both limit ceramide generation and to remove ceramide in order to become resistant to treatment. Furthermore, sphingosine kinase activity, which phosphorylates sphingosine the product of ceramide hydrolysis, has been linked to multidrug resistance, and can act as a strong survival factor. This review will examine several of the most frequently used cancer therapies and their effect on both ceramide generation and the mechanisms employed to remove it. The development and use of inhibitors of sphingosine kinase will be focused upon as an example of how targeting sphingolipid metabolism may provide an effective means to improve treatment response rates and reduce associated treatment toxicity. This article is part of a Special Issue entitled Tools to study lipid functions.     

Morphogenesis of the chick embryo limb. Competence of the embryonic and extra-embryonic ectoderm

Ecto-mesodermal interactions were investigated during the initiation of limb development in avian embryos. Experiments were performed on 2-day chick embryos. They consisted in implanting prospective leg mesoderm at different medio-lateral levels of the trunk and also into the extra-embryonic area. The implanted mesoderm was thus brought into contact with embryonic or extra-embryonic cicatricial or healing ectoderm, the ability of which to participate in the formation of an ectopic leg was tested. Whatever the level of embryonic ectoderm tested in hosts ranging from stage 14 to 27 pairs of somites (axial, paraxial, flank, ventrum), the experiments resulted in the formation of supernumerary limbs. Their frequency was level-dependent and decreased for each level, with increasing age of the host. The weakest competence was observed in the ectoderm of the prospective ventrum, the strongest in that of the prospective flank, axial and paraxial ectoderm showing an intermediary competence. Extra-embryonic ectoderm of blastoderms of the same age was unable to respond to the inducing action of the implanted prospective leg mesoderm. It was found to be incompetent, even at younger stages (5 to 13 pairs of somites).     

Étude autoradiographique de l'influence de la température sur la prolifération cellulaire chez les embryons âgés dePleurodeles waltlii Michah. (Amphibien,Urodéle)

Résumé Chez l'embryon de Pleurodèle au stade 34, la durée du cycle cellulaire et de ses phases varie peu selon les tissus mais dépend étroitement de la température. Le temps de génération et la durée de la phase S sont environ 3 ou 4 fois plus longs à 12° C qu'à 26° C. Lorsque la température s'élève, la phaseG ₂ est abrégée dans les mêmes proportions que la phaseM; par contre, la durée de la phaseG ₁ qui est nulle à 12° C s'allonge considérablement pour représenter environ 1/4 de la durée totale du cycle cellulaire à 26° C. La durée de cette phase est d'autant plus longue, à une température donnée, que les cellules sont plus différenciées. Les tissus étudiés représentent des populations cellulaires en croissance exponentielle. Le coefficient de prolifération, duquel dépend la base de la fonction exponentielle de croissance, est indépendant de la température mais particulier à chaque tissu. Il est d'autant plus faible que le tissu est plus différencié. En revanche, la vitesse de multiplication des cellules, qui est inversement proportionnelle au temps de génération, varie largement en fonction de la température; en outre, elle semble déterminer à elle seule la vitesse du développement des embryons aux températures choisies.

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Autoradiographic study of the effect of temperature on cellular proliferation in late embryos ofPleurodeles waltlii Michah. (Amphibia, Urodela)

Summary We observed in Pleurodeles embryos, stage 34, that the duration of the cell cycle and its phases was approximately the same for every tissue but was easily modified by varying the temperature. The generation time and the duration of S phase in embryos submitted to a 12° C temperature instead of 26° C are tripled or quadrupled. A temperature rise produced a proportionale shortening inG ₂ andM phases and a lengthening inG ₁ phase. ThisG ₁ phase is not detectable at 12° C but represent a 1/4 of the total generation time at 26° C. The more differentiated the cells are, the longer is theG ₁ time. The cell population studied during these experiments are growing exponentially. Growth fraction, which represents the exponential growth basis, is temperature independent but has a tissue specificity. This growth fraction is smaller the more the tissue is differentiated. However, the relative rate of cell division, inversely proportional to the generation time, is temperature dependent and appears to control the embryo's relative rate of growth under different temperatures.

     

Detection of bladder cancers using a SAMBA 200 cell image processor

The cell image analysis of urinary sediments was performed using a SAMBA 200 system. Cell profiles were created using 18 parameters related to size, shape, densitometry and chromatin texture. Learning sets of about 50 cell images per class were constructed for bening, degenerated benign, atypical, malignant and degenerated malignant urothelial cell types as well as for squamous epithelial and white blood cell types. A four-level hierarchic decision tree involving a discriminant analysis at each node was designed and then evaluated against a test set of 700 cells from the various classes. All of the cell images involved in this study were acquired from Papanicolaou-stained specimens obtained for routine screening. In spite of some misclassification errors, the analysis of the occurrence of cells in the various classes, especially the percentage of cells classified as suspicious (both atypical and malignant cells), by the SAMBA 200 system resulted in the separate clustering of the positive specimens (49 carcinomas grade II and higher) and the negative ones (26 benign samples). The preliminary results suggest that the cell population features (occurrence rate of cells in the various classes and mean cell profile within a class) may be of diagnostic value in designing a classifier dedicated to the prescreening of urinary sediments for the detection of bladder cancers.