Mise en place du PER en sciences de la nature dans l'espace BEJUNE

L'objet de cette recherche est de décrire l'appropriation et l'usage du PER dans l'enseignement des sciences de la nature, et de tirer de cette description une appréciation du changement que l'introduction de ce plan d'études a pu produire, ou du moins amorcer. Les vaines tentatives d'accès au travail effectif des enseignants nous ont conduits à leur proposer un bref questionnaire de 8 items. Sur un total de 302 envois, 176 questionnaires ont été retournés, dont 132 remplis de façon complète. D'une façon générale, les effets déclarés du PER semblent pour l'instant plutôt faibles, une des préoccupations prioritaires de la majorité des enseignants étant apparemment de disposer d'un programme de contenus enseignables, ce que le PER n'est pas, contrairement aux MER (moyens d'enseignement romands) attendus. (www.hep-bejune.ch)

Interaction between the fission yeast nim1/cdr1 protein kinase and a dynamin-related protein

AbstractThe nim1/cdr1 protein kinase is required for an efficient adaptation of cell cycle parameters to changes in nutritional conditions. We have isolated msp1, a new fission yeast member of the dynamin-related large GTPase family, in a two-hybrid screen designed to identify proteins interacting with the nim1 kinase. Msp1 has been shown to be essential for the maintenance of mtDNA and hence for the inheritance of functional mitochondria. We present evidence indicating that nim1 and msp1 proteins physically interact both in vitro and in vivo in fission yeast. These interactions occur through the amino-terminal catalytic domain of nim1 and the carboxy-terminal putative regulatory domain of msp1. These results provide new evidence for the existence of a connection between mitochondrial function and the cell cycle machinery

High-resolution in-depth imaging of optically cleared thick samples using an adaptive SPIM

Today, Light Sheet Fluorescence Microscopy (LSFM) makes it possible to image fluorescent samples through depths of several hundreds of microns. However, LSFM also suffers from scattering, absorption and optical aberrations. Spatial variations in the refractive index inside the samples cause major changes to the light path resulting in loss of signal and contrast in the deepest regions, thus impairing in-depth imaging capability. These effects are particularly marked when inhomogeneous, complex biological samples are under study. Recently, chemical treatments have been developed to render a sample transparent by homogenizing its refractive index (RI), consequently enabling a reduction of scattering phenomena and a simplification of optical aberration patterns. One drawback of these methods is that the resulting RI of cleared samples does not match the working RI medium generally used for LSFM lenses. This RI mismatch leads to the presence of low-order aberrations and therefore to a significant degradation of image quality. In this paper, we introduce an original optical-chemical combined method based on an adaptive SPIM and a water-based clearing protocol enabling compensation for aberrations arising from RI mismatches induced by optical clearing methods and acquisition of high-resolution in-depth images of optically cleared complex thick samples such as Multi-Cellular Tumour Spheroids

Structure tensor based analysis of cells and nuclei organization in tissues

International audienceMotivation: Extracting geometrical information from large 2D or 3D biomedical images is important to better understand fundamental phenomena such as morphogenesis. We address the problem of automatically analyzing spatial organization of cells or nuclei in 2D or 3D images of tissues. This problem is challenging due to the usually low quality of microscopy images as well as their typically large sizes. Results: The structure tensor is a simple and robust descriptor that was developed to analyze textures orientation. Contrarily to segmentation methods which rely on an object based modelling of images, the structure tensor views the sample at a macroscopic scale, like a continuum. We propose an original theoretical analysis of this tool and show that it allows quantifying two important features of nuclei in tissues: their privileged orientation as well as the ratio between the length of their main axes. A quantitative evaluation of the method is provided for synthetic and real 2D and 3D images. As an application, we analyze the nuclei orientation and anisotropy on multicellular tumor spheroids cryosections. This analysis reveals that cells are elongated in a privileged direction that is parallel to the boundary of the spheroid. Availability: Source codes are available at http://www.math.univ-toulouse.fr/~weiss

Multicellular tumor spheroid model to evaluate spatio-temporal dynamics effect of chemotherapeutics: application to the gemcitabine/CHK1 inhibitor combination in pancreatic cancer

International audienceUNLABELLED: ABSTRACT: BACKGROUND: The multicellular tumor spheroid (MCTS) is an in vitro model associating malignant-cell microenvironment and 3D organization as currently observed in avascular tumors. METHODS: In order to evaluate the relevance of this model for pre-clinical studies of drug combinations, we analyzed the effect of gemcitabine alone and in combination with the CHIR-124 CHK1 inhibitor in a Capan-2 pancreatic cell MCTS model. RESULTS: Compared to monolayer cultures, Capan-2 MCTS exhibited resistance to gemcitabine cytotoxic effect. This resistance was amplified in EGF-deprived quiescent spheroid suggesting that quiescent cells are playing a role in gemcitabine multicellular resistance. After a prolonged incubation with gemcitabine, DNA damages and massive apoptosis were observed throughout the spheroid while cell cycle arrest was restricted to the outer cell layer, indicating that gemcitabine-induced apoptosis is directly correlated to DNA damages. The combination of gemcitabine and CHIR-124 in this MCTS model, enhanced the sensitivity to the gemcitabine antiproliferative effect in correlation with an increase in DNA damage and apoptosis. CONCLUSIONS: These results demonstrate that our pancreatic MCTS model, suitable for both screening and imaging analysis, is a valuable advanced tool for evaluating the spatio-temporal effect of drugs and drug combinations in a chemoresistant and microenvironment-depending tumor model

Deep and Clear Optical Imaging of Thick Inhomogeneous Samples

Inhomogeneity in thick biological specimens results in poor imaging by light microscopy, which deteriorates as the focal plane moves deeper into the specimen. Here, we have combined selective plane illumination microscopy (SPIM) with wavefront sensor adaptive optics (wao). Our waoSPIM is based on a direct wavefront measure using a Hartmann-Shack wavefront sensor and fluorescent beads as point source emitters. We demonstrate the use of this waoSPIM method to correct distortions in three-dimensional biological imaging and to improve the quality of images from deep within thick inhomogeneous samples

Live cell division dynamics monitoring in 3D large spheroid tumor models using light sheet microscopy

<p>Abstract</p> <p>Background</p> <p>Multicellular tumor spheroids are models of increasing interest for cancer and cell biology studies. They allow considering cellular interactions in exploring cell cycle and cell division mechanisms. However, 3D imaging of cell division in living spheroids is technically challenging and has never been reported.</p> <p>Results</p> <p>Here, we report a major breakthrough based on the engineering of multicellular tumor spheroids expressing an histone H2B fluorescent nuclear reporter protein, and specifically designed sample holders to monitor live cell division dynamics in 3D large spheroids using an home-made selective-plane illumination microscope.</p> <p>Conclusions</p> <p>As illustrated using the antimitotic drug, paclitaxel, this technological advance paves the way for studies of the dynamics of cell divion processes in 3D and more generally for the investigation of tumor cell population biology in integrated system as the spheroid model.</p

Microcephalin and pericentrin regulate mitotic entry via centrosome-associated Chk1

Primary microcephaly, Seckel syndrome, and microcephalic osteodysplastic primordial dwarfism type II (MOPD II) are disorders exhibiting marked microcephaly, with small brain sizes reflecting reduced neuron production during fetal life. Although primary microcephaly can be caused by mutations in microcephalin (MCPH1), cells from patients with Seckel syndrome and MOPD II harbor mutations in ataxia telangiectasia and Rad3 related (ATR) or pericentrin (PCNT), leading to disturbed ATR signaling. In this study, we show that a lack of MCPH1 or PCNT results in a loss of Chk1 from centrosomes with subsequently deregulated activation of centrosomal cyclin B–Cdk1

Learning the cell cycle with a game: Virtual experiments in cell biology

Cell Cycle Learn (CCL) is a learning game designed for undergraduate students in Biology to learn common knowledge about the cell-division cycle along with practical skills related with setting up an experiment and the scientific method in general. In CCL, learners are guided through the process of formulating hypotheses, conducting virtual experiments and analysing the results in order to validate or invalidate the hypotheses. The game has been designed in the University of Toulouse and introduced last year as part of the curriculum of a cellular biology class. This paper presents early results of an evaluation of the game enabled by questionnaires filled by the participants and game data collected during the training sessions. The results demonstrate with examples that both types of data can be used to assess the game's utility