No description available.

No description available.

No description available.

Cotton fiber has been postulated to undergo a process of programmed cell death (PCD) during the maturation phase of development. A parallel may exist between cotton fibers and xylem tracheary elements, which have periods of elongation, secondary cell wall deposition and death. Secondary wall formation and PCD are purported to be coupled events in tracheary elements. In this study, an attempt was made to observe the occurrence and timing of PCD in cotton fibers by TUNEL staining to detect DNA strand breaks, and also to monitor DNA content by PI staining. The staining patterns produced by PI and TUNEL left room for interpretation. TUNEL-positive and PI-stained areas were observed, but failure to observe nuclei of conventional appearance in my cytological preparations at any time-point, along with possible nonspecific staining or autofluorescence of cell wall and intracellular components, made it difficult to draw firm conclusions of significance. Thus, additional analyses will be needed to prove or disprove current PCD theories. Nevertheless, the differences in TUNEL and PI signals across fiber development stages indicate that the observed fluorescence patterns are marking discrete developmental phases. The PI signal is dispersed throughout the cell during the elongation phase (5-15 DPA) and appears to condense during secondary cell wall synthesis (25- 40 DPA). TUNEL-positive signal may be observed as early as 25 DPA, but the signal is not widespread until 45 DPA. At 50 DPA and beyond, PI staining is reduced. Visually detectable DNA can be extracted from cotton fiber nuclei between 5 and 40 DPA, although a laddering pattern was not visible at any time-point. The results, although inconclusive, point to the possibility that PCD may be a process leading to maturation in the cotton fiber, succeeding completion of secondary cell wall synthesis.

Cotton fiber has been postulated to undergo a process of programmed cell death (PCD) during the maturation phase of development. A parallel may exist between cotton fibers and xylem tracheary elements, which have periods of elongation, secondary cell wall deposition and death. Secondary wall formation and PCD are purported to be coupled events in tracheary elements. In this study, an attempt was made to observe the occurrence and timing of PCD in cotton fibers by TUNEL staining to detect DNA strand breaks, and also to monitor DNA content by PI staining. The staining patterns produced by PI and TUNEL left room for interpretation. TUNEL-positive and PI-stained areas were observed, but failure to observe nuclei of conventional appearance in my cytological preparations at any time-point, along with possible nonspecific staining or autofluorescence of cell wall and intracellular components, made it difficult to draw firm conclusions of significance. Thus, additional analyses will be needed to prove or disprove current PCD theories. Nevertheless, the differences in TUNEL and PI signals across fiber development stages indicate that the observed fluorescence patterns are marking discrete developmental phases. The PI signal is dispersed throughout the cell during the elongation phase (5-15 DPA) and appears to condense during secondary cell wall synthesis (25- 40 DPA). TUNEL-positive signal may be observed as early as 25 DPA, but the signal is not widespread until 45 DPA. At 50 DPA and beyond, PI staining is reduced. Visually detectable DNA can be extracted from cotton fiber nuclei between 5 and 40 DPA, although a laddering pattern was not visible at any time-point. The results, although inconclusive, point to the possibility that PCD may be a process leading to maturation in the cotton fiber, succeeding completion of secondary cell wall synthesis.

No description available.

No description available.

No description available.

Programmed cell death (PCD) is a highly evolved mechanism of cellular suicide that is aberrantly activated following neural injury. Two fundamental PCD signaling pathways termed the extrinsic (caspase-8-mediated) and intrinsic (caspase-9-mediated) pathways, have been described. While each pathway is initiated by distinct cellular stimuli, both pathways culminate in the activation of downstream executioner caspases. Previous efforts to isolate the in vivo contribution of each pathway have been hindered by the embryonic lethality of casp8 and casp9 null mice. In the present study, I overcame this obstacle to directly assess the contribution of each pathway following two well-characterized forms of acute neural injury; excitotoxic destruction of CA1 pyramidal neurons, and the loss of motor neurons following facial nerve transection. To determine the role of caspase-8, I constructed several lines of mice in which caspase-8 was conditionally ablated within the relevant neuronal populations. The results obtained from these animals definitively demonstrate that caspase-8 is not required by either motor neurons or CA1 pyramidal neurons to undergo PCD following injury. Therefore, these findings have provided the first direct experimental evidence to counter the widely held dogma of caspase-8 as the central effector of death receptor-mediated signaling within neurons. With respect to the intrinsic pathway, several lines of evidence suggest that the apoptosome predominantly regulates the death of motor neurons. I tested this hypothesis by performing facial axotomies in mice containing a point mutation introduced (“knocked in”) into the genomic locus of cytochrome c which abolishes its ability to activate the intrinsic pathway. Homozygous cytochrome c knock-in mice displayed a significant enhancement in motor neuron survival in comparison to control littermates following injury. However, the level of motor neuron protection differed from that previously reported in mice either overexpressing anti-apoptotic or lacking pro-apoptotic members of the Bcl-2 family. Therefore, the results of this study directly demonstrate the influence of the apoptosome on injury-induced neuronal PCD isolated from upstream Bcl-2 family-mediated effects. In addition, my results have provided the first evidence that activation of the apoptosome is required for the release of apoptosis inducing factor (AIF) from the mitochondria of injured motor neurons in vivo.

Introduction : l’infection par le cytomégalovirus (CMV) humain est la plus fréquente des infections après greffe d'organe. Des effets indirects à long terme sont fortement suspectés mais restent encore largement incompris. Notre travail de thèse s’est intéressé à mieux comprendre les conséquences de l’infection du donneur par le CMV sur la réponse immunitaire du receveur et sur le devenir de son greffon. Résultat : nous avons initialement rapporté que l'infection du donneur (D+) par le CMV est un facteur de risque indépendant de perte de fonction du greffon rénal particulièrement si le receveur est également séropositif avant la greffe (D+R+ comparé aux D+R-). Le risque est fortement majoré en cas de mésappariement complet en HLA de classe I entre le receveur et son donneur. Puis nous avons analysé le rôle du greffon infecté dans le développement de la réponse lymphocytaire anti-CMV. Nous avons rapporté pour la première fois que la superinfection CMV entraine une augmentation du nombre de LT CD8 répondeurs spécifiques du CMV à distance de la transplantation, à condition que le donneur et le receveur partagent des identités HLA-I. De plus nous avons montré chez le sujet D+R- que l'expansion des lymphocytes T CD8 anti CMV restreints par le HLA-A2 nécessite l'expression de ce HLA par le donneur. Ces résultats ensemble indiquent le rôle des cellules du donneur dans l’inflation des LT CD8 anti-CMV à distance de la greffe. Dans un troisième travail, nous avons montré qu’un polymorphisme du gène de Programmed Cell Death 1 (PD-1.3) influe sur la survie des greffons rénaux et pulmonaires D+, les patients porteurs de l’allèle variant A ayant un meilleur pronostic que les patients homozygotes GG. Nos données indiquent aussi que les patients homozygotes AA ont un plus grand nombre de lymphocytes anti-CMV producteurs d'IFN-ɣ, suggérant que ce polymorphisme pourrait être associé à une dysfonction de la réponse immunitaire spécifique anti-CMV. Conclusion : ensemble ces données suggèrent pour la première fois que la qualité de la réponse lymphocytaire cytotoxique anti-CMV pourrait être importante pour contrôler la réplication virale dans le greffon et les lésions induites par cette dernière. Ainsi nous proposons deux mécanismes à l’origine du développement des lésions liées à l'infection à CMV dans le rein: défaut de reconnaissance des cellules allogéniques infectées en cas de mésappariement complet en HLA de classe I et une dysfonction LT CD8 anti-CMV. / Background: cytomegalovirus (CMV) is the leading cause of viral infection after solid organ transplantation. Despite a large body of literature, the effects of chronic cytomegalovirus (CMV) infection on graft outcome remain controversial.Results: we first reported that donor CMV infection (D+) was an independent risk factor of kidney graft loss, especially in pretransplant infected recipients (R+). In addition, we observed that full HLA-I mismatching was an important determinant of this risk. In a second study, we focused on effect of donor CMV infection on anti-CMV specific immune response. We reported that CMV superinfection greatly increased the number of anti-CMV IFN-ɣ-producing T cells, provided that donor and recipient shared at least one HLA-I identity. Then in D+R- HLA-A2-expressing recipients, we compared the number of anti-CMVpp65 CD8+T cells restricted by HLA-A2 depending on whether the donor expressed or not HLA-A2. Patients who received non-HLA-A2 kidneys developed very few anti-CMVpp65 T-cells restricted by HLA-A2 as compared to those who received an HLA-A2-expressing kidney. This result indicated that presentation of CMV peptides by donor cells was crucial to stimulate the expansion of pp65-specific memory CD8 T cells. Finally, we established that a SNP in the Programmed Cell Death 1 gene (PD-1.3) influenced D+ kidney and lung transplants survival, while it was also associated with the level of anti-CMV specific T-cell response. Conclusion: taken together, these data suggest that anti-CMV specific immune response is pivotal to control infection within the graft and prevent subsequent organ damages. We propose two mechanisms to explain effect of donor CMV infection on graft outcome: (1) inability of anti-CMV CD8 T cells to recognize donor-infected cells in case of full HLA-I mismatching, (2) dysfunction of anti-CMV CD8 T cells after transplantation in some patients, highlighted by our genetic study.