The interaction of a cell with its environment occurs through the plasma membrane. To regulate the shape and composition of their plasma membranes, and to internalize nutrients and signaling factors, eukaryotic cells use endocytosis, the formation of vesicles from the plasma membrane. Clathrin mediated endocytosis (CME) is the most widely used form of endocytosis. The dynamics and molecular mechanisms of CME, and its role in a number of cellular functions, has been the subject of intense study. A vast number of proteins and interactions implicated in this process have been identified, but the dynamics of these interactions is largely unknown. Our interdisciplinary team of biologists and chemists will develop new tools to visualize and interfere with selected key interactions leading to clathrin coated vesicle (CCV) formation. First, we will synthesize photoactivatable, or caged, peptides to block the interaction between the proline rich domain of dynamin, a protein essential for CME, and the SH3 (Src homology 3) domain of its partners, such as amphiphysin. This will block CME in live cells in a spatially and temporally controlled way, which we will test with live cell imaging of CCV formation. Second, we will construct pairs of endocytic proteins fused with fluorescent proteins to visualize their interaction by Förster Resonance Energy Transfer-Fluorescence LIfetime Microsopy (FRET-FLIM) imaging in live cells. We will connect the timing of this interaction to the stages of endocytic vesicle creation. This set of experiments will provide unprecedented details on the dynamics of molecular mechanisms of CME. Moreover, the caged peptides developed in this project will be usable in a variety of cellular contexts. In particular, CME is essential for the expression of synaptic long term depression (LTD), a neuronal substrate of certain forms of learning and memory. We will perform electrophysiology patch clamp experiments to determine with the caged peptide where and when CME is required for LTD, and whether endocytosis contributes to the synapse specificity of LTD. We anticipate that these tools, once made available to the scientific community, will clarify the involvement of CME in a number of cellular functions.

Voice production is the result of a complex interaction between the air expelled from the lungs, the vocal tract from the larynx to the lips and the vocal folds delimiting the glottis. The pulsatile flow generated during the self-sustained oscillations of the glottis wall tissues acts as an source for acoustic waves that will carry part of the speech information. The vocal folds behaves as a valve that periodically opens and closes the glottis, both modulating the airflow and sustained by the fluid-structure energy transfer. The existence of these oscillations is however only possible due to the jet formed at the glottal exit and by suitable vocal folds characteristics. These latter are constituted by several tissues layers that are controlled, consciously or not, by the ligament tension and the muscular activity, and subject to the action of the pressure force from the air flowing through the glottis. This complex heterogeneous structure may be degraded, leading to potential source of voice disorders. The objective of the VOFOCAM project is to develop the tools that make possible to observe and analyse the structural health of the laryngeal tissues (vocal folds and ventricular bands). Vocal folds mechanical replicas as the one of the GIPSA-lab (UMR 5216, CNRS – Univ. Grenoble) have been set up to examine under controlled conditions the peculiarities of the glottal flow and its interaction with mechanical deformable structures and with the acoustic resonant vocal tract. The replicas provide a support for the development of an measurement device to capture the large deformation of the artificial vocal folds. Using a wide range of material, tension and positioning, we aim to built a large set of configurations that would be as representative as possible of the laryngeal pathologies that will analysed with mathematical methodologies accounting for the spatially distributed information. The reduced order model that could in particular represent anterior-posterior, left-right and subglottal-supraglottal asymmetries will induce the parametrization of the vibration patterns observed on the oscillating replica, and allow to build a database suitable for sound-synthesis purposes and that could be the basis of a human vocal folds analysis device in conjunction with the transposition of the developed experimental techniques in a clinical context.

In 2003, the French Ministry of Research expressed the need for a national initiative in nanosciences and nanotechnologies in order to increase the competitiveness of the national research community in this field. Indeed, it is generally accepted that the ability to model, image, manipulate matter at micro and nanoscale is leading to new technologies that impact many sectors of the economy including information and communications, healthcare, transportations, energy and security. The Ministry launched with the “Centre National de la Recherche Scientifique (CNRS)” and the “Commissariat à l'énergie atomique et aux énergies alternatives (CEA2)”, a dedicated programme entitled "A national network of large technological facilities and Basic Technological Research in micro and nanotechnologies" to be named as the RTB programme throughout this document. The RTB programme covered three main fields: -The development of competitive clean-room infrastructures, able to provide the support and technologies needed for the national research activities of the national community in the field of micro and nanotechnologies; -The training of PhDs and post-docs in technological facilities; -The production of IP activity in nanotechnology for future exploitation through industry. It involves a network of 6 academic (CNRS and universities) facilities distributed over France belonging to the “RENATECH” network (www.renatech.org) and associated to a large facility, CEA-Leti (www.leti.fr), located in Grenoble and dedicated to technological developments targeting transfer to industry. Mainly supported by an investment plan, the results of the RTB programme are reviewed each year by an international panel on the basis of the leverage for the scientific and industrial community and the progress of the networking activity.

The use of nanomedicine has led to significant success in drug targeting to specific diseased tissues and cells. Several impediments of nanocarriers still limit a wider use of nanomedecine: low drug loading and uncontrolled release, poor versatility and scalability, difficulty to include imaging contrast agents to achieve ‘theranostic’ properties. These challenges are particularly acute for the delivery of nucleotides and their analogues, despite their high and widespread pharmacological activities. We aim at designing a novel drug delivery platform tailored to these hydrophilic drugs, displaying contrast agent properties and scalability potential. This platform consists in organic/inorganic nanogels based on chitosan coupled to metal (Fe, Zn, Gd) or metal complexes. Such nanogels will be investigated for their potential to deliver nucleotides and their analogues to target cells and tissues, and enhance contrast in magnetic resonance imaging. The potential of this theranostic platform to be scaled-up will also be investigated.

In 2003, the French Ministry of Research expressed the need for a national initiative in nanosciences and nanotechnologies in order to increase the competitiveness of the national research community in this field. Indeed, it is generally accepted that the ability to model, image, manipulate matter at micro and nanoscale is leading to new technologies that impact many sectors of the economy including information and communications, healthcare, transportations, energy and security. The Ministry launched with the “Centre National de la Recherche Scientifique (CNRS)” and the “Commissariat à l'énergie atomique et aux énergies alternatives (CEA2)”, a dedicated programme entitled "A national network of large technological facilities and Basic Technological Research in micro and nanotechnologies" to be named as the RTB programme throughout this document. The RTB programme covered three main fields: ? The development of competitive clean-room infrastructures, able to provide the support and technologies needed for the national research activities of the national community in the field of micro and nanotechnologies; ? The training of PhDs and post-docs in technological facilities; ? The production of IP activity in nanotechnology for future exploitation through industry. It involves a network of 6 academic (CNRS and universities) facilities distributed over France belonging to the “RENATECH” network (www.renatech.org) and associated to a large facility, CEA-Leti (www.leti.fr), located in Grenoble and dedicated to technological developments targeting transfer to industry. Mainly supported by an investment plan, the results of the RTB programme are reviewed each year by an international panel on the basis of the leverage for the scientific and industrial community and the progress of the networking activity.