Riassunti dei progetti del corso di dottorato di ricerca in biologia molecolare e cellulare
Project leader: ELISABETTA CAPORALI (elisabetta.caporali@unimi.it)
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- Project leader: GRAZIELLA CAPPELLETTI ( graziella.cappelletti@unimi.it )
- RESEARCH PROJECT SUMMARY
- Project leader: GIUSEPPINA CARETTI (giuseppina.caretti@unimi.it)
Project leader: ELISABETTA CAPORALI (elisabetta.caporali@unimi.it)Location: Department of Biosciences, University of Milan, ItalyRESEARCH PROJECT SUMMARYKey cell wall modelling factors to design fruit shapes in Arabidopsis] Morphogenesis is the remarkable process by which a developing plant acquires its shape. Underlying the architectural complexity of plants are diverse cell types that easily reveal relationships between cell structure and specialized functions. Much less obvious are the mechanisms by which the cellular growth machinery and mechanical properties of the cell wall interact to dictate cell shape that lead to an organ formation. Fully developed fruits have a complex primary cell wall matrix, and are exposed to a highly precise regulation required to determine the final size and shape of the organ. Although progress is being made in identifying and characterizing the genes required for the synthesis of fruit cell wall matrix components, little is known about how the production and accumulation of wall components are regulated at different levels: transcriptional control, biochemical control, or both. It remains a central challenge for developmental biology how much this regulation contributes to produce the diversity of cell shapes and functions that characterize the formation of a seed or a silique. We propose an innovative investigation of morphogenesis to address how these elements are controlled at the molecular and cell level, and how the mechanical properties of these elements lead to specific growth patterns using seed and fruit development in Arabidopsis as a model system. The outcome of this project not only will discuss the unique geometric properties and physical processes that regulate seed and fruit organogenesis in Arabidopsis thaliana, but also will lead us to develop testable mathematical models that improve our understanding of how genetic networks, protein motors, and extracellular polymer properties of these elements lead to specific seed/fruit growth patterns.
Project leader: GRAZIELLA CAPPELLETTI (graziella.cappelletti@unimi.it)Location: Department of Biosciences, University of Milan, ItalyCo-Tutor: Francesco Demartin Dipartimento di Chimica, Via Venezian 21, Milano RESEARCH PROJECT SUMMARYDissecting microtubule dynamics at the synapse Microtubules (MTs) are highly dynamic polymers that control many aspects of neuronal function: they provide a scaffold to sustain axonal and dendritic architecture and supply the railway for axonal transport. Far from being mere structural elements, MTs are emerging as key determinants of neuronal polarity. In spite of the fact that the regulation of MT organization and dynamics has been extensively studied during axon and dendrite formation and maintenance, much less is known about the regulation of MT dynamics at synaptic terminals. Recently, we have unravelled the role of a synaptic protein, namely -synuclein, in regulating MTs. The goal of the present project is to investigate the role of -synuclein in controlling MT behaviour at the synapse by focusing on MT nucleation that determines where, when and how polymerization of new MTs is initiated. In the first part of the work, the PhD fellow will perform light and electron microscopy analyses on primary neuronal cultures obtained from mice embryos. In these cultures, -synuclein will be overexpressed or knocked-down. Next, taking advantage of the very high resolution afforded by 3D EM tomography, a detailed analysis of the structure of MTs nucleated at the synapse will be carried out. The labelling with immuno-gold particle will allow localizing -synuclein into the 3D structures. For image analysis will be used a software developed by the Department of Chemistry (University of Milan). A better understanding of MT regulation at the synapse and novel insights into the physiological role of -synuclein is the expected outcome.
Location: Department of Biosciences, University of Milan, ItalyRESEARCH PROJECT SUMMARYEpigenetic mechanisms underlying skeletal muscle atrophy We are interested in studying the epigenetic regulation of factors involved in skeletal muscle wasting. Muscle wasting occurs in association with different conditions such as atrophy of disuse, muscular dystrophies, sarcopenia of aging, and cachexia secondary to other diseases (as cancer, heart diseases or chronic obstructive lung disease). In all these circumstances, muscle wasting manifests with overlapping features and similar molecular mechanisms (Ruegg and Glass, 2010). One of the key factors involved in muscle wasting is myostatin, which is a negative regulator of skeletal muscle mass and is up-regulated in atrophic conditions. Myostatin loss of function in knockout mice or in naturally occurring mutants leads to a significant increase in muscle mass, known as “double muscling” (Huang et al., 2011). We have recently shown that the histone-methylase SMYD3 and the bromodomain protein BRD4 positively regulate transcription of the myostatin gene, both in muscle homeostasis and during muscle atrophy. Importantly, myostatin levels can be reduced in in vitro cultured myotubes by a recently developed epigenetic drug, called JQ1, which associates with the bromodomains of BRD4 and blocks its function on the chromatin (Proserpio et al., 2013). The specific aims of the project are: 1) to study the ability of JQ1 to block myostatin and muscle loss in vivo, using different models of muscle wasting. 2) To investigate the effect of the small inhibitor JQ1 on the function of adult skeletal muscle stem cells, also known as satellite cells. 3) To identify the genes altered by BRD4 blockade and by SMYD3 reduction in normal skeletal muscle and in atrophic conditions. References: Ruegg MA and Glass DJ. Molecular Mechanisms and Treatment Options for Muscle Wasting Diseases. Annu. Rev. Pharmacol. Toxicol. 2011.51:373-395 Huang Z, Chen X, Chen D. Myostatin: a novel insight into its role in metabolism, signal pathways, and expression regulation. Cell Signal 2011;23(9):1441-6. Proserpio V., Fittipaldi R., Ryall J.G, Sartorelli V., Caretti G. The Methyltransferase SMYD3 Mediates the Recruitment of Transcriptional Elongation Factors at the Myostatin and c-Met Genes and Regulates Skeletal Muscle Atrophy. Genes & Development. 2013; 27; 1299-1314 Project leader: ELENA CATTANEO (elena.cattaneo@unimi.it; cattaneolab@unimi.it) Location: Department of Biosciences (Via Viotti 3, University of Milan, Italy RESEARCH PROJECT SUMMARY hES cells differentiation into striatal neurons by conditional over-expression of transcription factors Huntington’s disease (HD) is an autosomal-dominant, progressive neurodegenerative disorder that usually onsets in midlife. It is characterized by motor, cognitive, and psychiatric symptoms. Once symptomatic, patients are rapidly disabled and require increasing multidisciplinary care. HD is a tremendous burden for medical, social, and family resources. The symptoms and the progression of HD can be linked to its neuropathology, which is characterized by loss of specific neuronal populations in many brain regions. Several studies have shown that medium spiny neurons (MSN) are severely affected in HD. MSN are inhibitory projection neurons and are the primary source of striatal projections. The laboratory is actively involved in international research programmes aiming at deriving specific and robust differentiation protocols for the generation of MSN. Most recently we have developed a protocol to obtain such neurons from human embryonic stem (hES) or from induced pluripotent stem cells (hiPS) using a defined in vitro neural induction system and quantitative assessment tools (Delli Carri, 2013). In this project we aim at developing strategies to further improve the recovery and quality of fully functional human MSN from hES/hiPS cells with the goal of future transplantation studies in HD. We will combine morphogens treatment with transcription factors inducible over-expression. We plan to develop doxycycline-inducible hES lines that over-express critical combinations of transcription factors (TFs) known to be important for striatal specification and differentiation. Changes in gene transcriptional profiling and expression of positional markers will be used to verify the identity acquired by the implemented cells as they progress along neuronal differentiation. Cell sorting will be employed to further select for suitable neural progenitors. Quality of the neurons obtained at the end of the differentiation protocol will be verified by a convergence of features such as expression of neuronal markers as well as neurochemical and bioelectrical properties. In conclusion this project aims at (i) developing new hES cell lines over-expressing critical striatal TFs (ii) characterizing the identity of the neural progenitors and post-mitotic neurons derived from differentiation studies. Moreover, the project will also focus on refining existing protocols for making striatal neurons, by considering developing cell sorting strategies and small molecules treatments. Download 336.57 Kb. Share with your friends:
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