Les approches complémentaires à l’expérimentation animale en agronomie et clinique vétérinaire : Solutions et limites

En France, l’utilisation des animaux à des fins scientifiques fait l’objet d’une réglementation stricte depuis plus de 25 ans. Son évolution s'effectue principalement dans un cadre européen. La directive 86/609 visait ainsi à l'harmonisation des pratiques entre les Etats membres. En France, comme en Europe, les textes réglementaires sur la protection animale et l’utilisation de l’animal en expérimentation sont de plus en plus exigeants. La directive 2010/63/UE du Parlement Européen et du Conseil du 22 septembre 2010 relative à la protection des animaux utilisés à des fins scientifiques a fortement renforcé les exigences vis-à-vis de l’utilisation des animaux. Cette nouvelle directive s'attache plus particulièrement aux mesures concernant l’évolution du nombre d’animaux utilisés à des fins scientifiques et éducatives, cette utilisation « demeurant nécessaire pour protéger la santé humaine, la santé animale et l'environnement ». Le corpus réglementaire fait de la diminution du nombre d’animaux en expérimentation un défi majeur de la société scientifique. Des méthodes modernes telles que les méthodes in vitro, in silico et de modélisation permettent actuellement de diminuer le nombre d’animaux en expérimentation animale et d’être complémentaires à ces expérimentations. Ces méthodes sont en plein essor et il reste encore de nombreuses découvertes à faire afin de pouvoir répondre à plus de questions scientifiques par des méthodes alternatives

The broad OVI absorber at z = 3.02 toward CTQ 325

We report on the multiphase absorption-line system detected at a redshift z = 3.021 in the spectrum of the quasi-stellar object CTQ 325 (z_em = 3.212). The system is displaced by ~14,000 km/s to the blue of the systemic velocity defined by the center of the symmetric OI quasar emission line. It consists of shallow and broad (~700 km/s, FWHM) absorption lines of HI 1215, CIV 1548,1550, and OVI 1032,1038, produced by collisionally ionized gas and of narrow absorption lines (FWHM < 20 km/s) of Ly-alpha, Ly-beta, ..., Ly-6, SiIII 1206, and CIII 977 produced by photoionized gas. Possible origins of this system are discussed.Comment: 6 pages, 4 figures, to appear in A&

Genomic DNA PCR analysis to assess xenograft development in mouse mammary gland

The mouse transplantation model remains the most relevant methodology to assess the functional capacities of mammary cells and is particularly appropriate for investigations regarding mammary stem cells, whatever the species studied. Following xenotransplantation in mice mammary fat pad, the development of the xenograft is commonly evaluated by immunohistology. Here, we present a simple and rapid method to control the species specificity of a xenograft based on genomic DNA PCR amplification. DNA is extracted from the fixed samples intended for histology, thus allowing the reuse of precious samples. Standard and digital droplet PCR (requiring low DNA quantities) methods have been used to make the present method suitable for the analysis of xenotransplanted samples

Développement des modèles de culture cellulaire de muscle en 3D : de nouvelles opportunités pour les productions animales

Le muscle squelettique est organisé en faisceau de fibres musculaires de différentes tailles et parcouru par des réseaux vasculaires et nerveux. Les cellules satellites sont des cellules souches logées le long des fibres musculaires et sont à la base des progéniteurs myogéniques (myoblastes). Les cellules satellites peuvent être aisément extraites du muscle et cultivées. Les modèles typiques de culture en deux dimensions (2D) de cellules dérivées du muscle squelettique ne peuvent pas recréer complètement l'organisation et la fonction des tissus musculaires vivants, ce qui limite leurs utilités dans les études physiologiques approfondies. Le développement de modèles de culture 3D fonctionnels offre une opportunité unique pour mimer les tissus vivants et modéliser les maladies musculaires. A cet égard, ce nouveau type de modèles in vitro augmente significativement notre compréhension de l'implication des différentes populations cellulaires dans la formation du muscle squelettique et de leurs interactions, ainsi que les modalités de réponse d'un muscle pathologique à de nouvelles thérapies. Ce deuxième point pourrait conduire à l'identification de traitements efficaces. Dans cette synthèse, nous traitons des progrès significatifs qui ont été réalisés ces dernières années pour concevoir des structures ressemblant à des tissus musculaires, fournissant des outils utiles pour étudier le comportement des cellules souches résidentes. Nous nous intéressons plus particulièrement au développement de systèmes basés sur des « myosphères » et des faisceaux de fibres ou « myobundles » ainsi que sur les systèmes de bio-impression. Les protocoles de stimulation électrique/mécanique et les systèmes de co-culture développés pour améliorer le processus et les fonctionnalités de maturation des tissus seront également présentés. La formation de tissus musculaires biomimétiques représente une nouvelle technologie pour étudier la fonction et l'organisation spatiale des muscles squelettiques dans un grand nombre de contextes physiologiques, pathologiques et agronomiques

Spectroscopic follow-up of FIRBACK-South-Marano bright galaxies

We present optical spectroscopy of the brightest 170 microns sources in the FIRBACK South Marano (FSM) field. The survey is 90% complete at the 4 sigma level. The spectra are compared with those of a reference sample of faint IRAS (60 microns selected) sources observed in the same conditions . The galaxies in both samples are characterized by a predominance of emission-line spectra and moderate IR luminosities (10E10.5 < L_IR < 10E12 L_Sun). The fraction of AGN's is low (about 15%) and the majority of sources are nearby (z<0.3), dusty, star forming galaxies, with moderate star formation rates (a few 10 solar masses per year). The infrared emission of the FSM galaxies shows also a colder spectral energy distribution than was expected. The galaxies in both samples (IRAS faint sources and FSM) are essentially undistinguishable with the available data, and seem to represent a population of closeby, cold, star forming galaxies rather neglected up to now. Although their contribution to the far-IR background seems to be low, they deserve a more detailed study on their own. The contribution of fainter, presumably more distant, galaxies to the far-IR background will be discussed when we will have completed the follow-up of the fainter part of this 170 microns survey. It seems however already clear that the fraction of closeby, cold galaxies is much larger than expected by the earlier models, at least down to the sensitivity achieved by this ISO survey.Comment: 16 pages + 9 figures (Figs 1, 8 and 9 too big to be included) Accepted in Astronomy & Astrophysic

A bovine lymphosarcoma cell line infected with theileria annulata exhibits an irreversible reconfiguration of host cell gene expression

Theileria annulata, an intracellular parasite of bovine lymphoid cells, induces substantial phenotypic alterations to its host cell including continuous proliferation, cytoskeletal changes and resistance to apoptosis. While parasite induced modulation of host cell signal transduction pathways and NFκB activation are established, there remains considerable speculation on the complexities of the parasite directed control mechanisms that govern these radical changes to the host cell. Our objectives in this study were to provide a comprehensive analysis of the global changes to host cell gene expression with emphasis on those that result from direct intervention by the parasite. By using comparative microarray analysis of an uninfected bovine cell line and its Theileria infected counterpart, in conjunction with use of the specific parasitacidal agent, buparvaquone, we have identified a large number of host cell gene expression changes that result from parasite infection. Our results indicate that the viable parasite can irreversibly modify the transformed phenotype of a bovine cell line. Fifty percent of genes with altered expression failed to show a reversible response to parasite death, a possible contributing factor to initiation of host cell apoptosis. The genes that did show an early predicted response to loss of parasite viability highlighted a sub-group of genes that are likely to be under direct control by parasite infection. Network and pathway analysis demonstrated that this sub-group is significantly enriched for genes involved in regulation of chromatin modification and gene expression. The results provide evidence that the Theileria parasite has the regulatory capacity to generate widespread change to host cell gene expression in a complex and largely irreversible manner

The protozoan parasite Theileria annulata alters the differentiation state of the infected macrophage and suppresses musculoaponeurotic fibrosarcoma oncogene (MAF) transcription factors

AbstractThe tick-borne protozoan parasite Theileria annulata causes a debilitating disease of cattle called Tropical Theileriosis. The parasite predominantly invades bovine macrophages (mϕ) and induces host cell transformation by a mechanism that has not been fully elucidated. Infection is associated with loss of characteristic mϕ functions and phenotypic markers, indicative of host cell de-differentiation. We have investigated the effect of T. annulata infection on the expression of the mϕ differentiation marker c-maf. The up-regulation of c-maf mRNA levels observed during bovine monocyte differentiation to mϕ was suppressed by T. annulata infection. Furthermore, mRNA levels for c-maf and the closely related transcription factor mafB were significantly lower in established T. annulata-infected cell-lines than in bovine monocyte-derived mϕ. Treatment of T. annulata-infected cells with the theileriacidal drug buparvaquone induced up-regulation of c-maf and mafB, which correlated with altered expression of down-stream target genes, e.g. up-regulation of integrin B7 and down-regulation of IL12A. Furthermore, T. annulata infection is associated with the suppression of the transcription factors, Pu.1 and RUNX1, and colony stimulating factor 1 receptor (CSF1R) which are also involved in the regulation of monocyte/mϕ differentiation. We believe these results provide the first direct evidence that T. annulata modulates the host mϕ differentiation state, which may diminish the defence capabilities of the infected cell and/or promote cell proliferation. Musculoaponeurotic fibrosarcoma oncogene (MAF) transcription factors play an important role in cell proliferation, differentiation and survival; therefore, regulation of these genes may be a major mechanism employed by T. annulata to survive within the infected mϕ

Etude des mécanismes de mort cellulaire programmée dans les cellules transformées

PARIS7-Bibliothèque centrale (751132105) / SudocSudocFranceF

<em>Technical note:</em> Quantification of caseins from a crude extract of mammary epithelial cells

International audienceEn masse secretion of milk proteins, notably the caseins in the form of casein micelles, is a unique feature of the milk-secreting mammary epithelial cell. Caseins are therefore specific markers of these cells and constitute an ideal tool to monitor their differentiation, as well as functional status, during the development of the gland. To use them as such, a reliable method for quantitative analysis of the caseins from mammary cells or tissue is needed. Here we show that the caseins are heat-stable, a feature that leads to their complete extraction from a complex cellular extract by boiling. This allows for high enrichment and direct analysis of the caseins, even when they are poorly expressed in the starting material