PhD projects




PhD projects

Title of PhD project (See details below)	Academic partner	Supervisor	ESRs	Secondment
Synaptic function and plasticity
Assessing the action on synaptic strength and plasticity of GluK1 overexpression in a Down’s syndrome animal model.	UMH	Juan Lerma	Wilfrid Mazier	Noscira
Dissection of the BOLD signal in fMRI using optogenetic tools.	UMH	Santiago Canals	Pierrick Jego	Noscira
Characterisation of GABAergic neurotransmission within the globus pallidus and subthalamic nucleus network in a rat model of Parkinson’s disease.	UBx2	Maurice Garret	Zhuowei Du	Explora Nova
The role of cholinergic receptors in synaptic plasticity	UniBris	Jack Mellor	Valeria Olivo	NeuroSearch
Role of the endocannabinoid system in morphological plasticity	UBx2	Valentin Nägerl & Giovanni Marsicano	Philipp Bethge	To be determined

Synaptic development and maturation
Regulation by mental retardation protein PAK3 of PSD formation in newly formed synapses	UniGe	Dominique Muller	Katalin Sari	Explora Nova Amplitude Systemes
Characterization of genes involved in mental retardation.	UMIL	Maria Passafaro	Jonathan Zapata	Aptuit
Functional characterization of a Scribble1-Vangl2-Lgl1 complex in the hippocampal network of new animal models of mental retardation	UBx2	Nathalie Sans	Vera Pinheiro	BioXtal

Synaptic molecular complexes and trafficking of synaptic proteins
Intracellular signaling mechanisms during ischaemia	UniBris	John Hanley	Elena Blanco Suarez	GSK
Mechanisms and neuroprotective properties of SUMOylation of synaptic protein in models of ischaemia.	UniBris	Jeremy Henley	Fernando Josa Prado	GSK
Analysis of the mechanisms of signaling scaffolds in synaptic plasticity	UniGö	Oliver Schlüter	Tanmoy Samaddar	Aptuit
Characterization of DA and Glu interaction in experimental parkinsonism	UMIL	Monica Di Luca	Jennifer Stanic	NeuroSearch
Molecular mechanisms regulating postsynaptic density protein assembly and plasticity	UMIL	Carlo Sala	Christopher Heise	Bioxtal
Regulation of nicotinic receptor trafficking and synaptic function in C. elegans.	UniGö	Stefan Eimer	Suekyoung Jeon
Development of a new technique for fast multiphoton super-resolution imaging of thick biological samples	UBx2	Daniel Choquet & Jean Baptiste Sibarita	Kalina Hass	Amplitude Systemes

Synapses in networks and higher brain function
Short-term plasticity and regulation of synaptic responses by activity under natural conditions	UMH	Miguel Maravall	Giovanni Ferrati	Noscira
Synaptic plasticity at hippocampal mossy fiber synapses from slices to in vivo	UBx2	Christophe Mulle	Stefano Zucca	NeuroSearch
Changes of GABAB inhibition in animal models of neuropathic pain	UBx2	Marc Landry & Alexandre Favereau	Sara Elramah	Explora Nova
Assessing changes in synaptic plasticity in perirhinal cortex	UniBris	Zafar Bashir	Medhi Bhouri	GSK
Channel Rhodopsin 2 as a tool to activate midbrain dopamine neurons	UniGe	Christian Lüscher	Tifei Yuan	Aptuit
	UniGe	Christian Lüscher	Sebastiano Bariselli	To be determined
Imaging of activity-related structural changes in synapses in vivo.	UniGe	Anthony Holtmaat	Vassilis Kehayas	Explora Nova Amplitude Systemes
Defects in neocortical circuits underlying cognitive disorders.	UBx2	Andreas Frick	Maria Szlapczynska	Femtonics

WP.1.1. Supervisor: Pr. Juan Lerma
Project title: Assessing the action of GluK1 overexpression on synaptic strength and plasticity in a Down’s syndrome animal model.
Objectives: The human gene coding for the glutamate receptor GluK1 (GRIK1) localizes in chromosome 21q22.1, close to the APP mutated gene in amyotrophic lateral sclerosis (ALS) and could be involved in neuronal death associated to ALS. Similarly, an excess of this receptor at the synapse may influence synaptic strength at several levels, in the spinal cord but also in the hippocampus and neocortex., A mouse model of Down’s syndrome, Ts65Dn mice, carries a partial duplication of chromosome 16 which includes the GRIK1 gene. Ts65Dn mice could be a good model to understand the role of GluK5 in synaptic transmission and its putative influence in Down’s phenotype. We will analyze the behaviour of synaptic responses in the spinal cord and the necortex under stimulation of dorsal roots, thalamocortical projection and intracortical cell populations. We will relate these actions to phenotypic characteristics of Down’s syndorme.
Methodology: In vitro patch clamp and extracellular recordings –spinal cord and brain slices-. Confocal an two-photon calcium imaging. Real time PCR.
Secondment : Noscira S.A.

Student's name : Mazier Wilfrid / Lab: Juan Lerma laboratory, Synaptic Physiology group, Instituto de Neurociencias de Alicante UMH-CSIC

Education: Master´s by reseach in Neurosciences and Neuropsychopharmacology

Origin: University of Bordeaux II, France

Project title : Assessing the action of GluK1 kainate receptor subunit overexpression on synaptic strengh and plasticity in a Down syndrome animal model.

The human gene coding for the glutamate receptor GluK1 (GRIK1) localizes in chromosome 21q22.1, close to the APP mutated gene in amyotrophic lateral sclerosis (ALS) and could be involved in neuronal death associated to ALS. Similarly, an excess of this receptor at the synapse may influence synaptic strength at several levels, in the hippocampus but also in the spinal cord and neocortex., A mouse model of Down's syndrome, Rb(12.Ts171665Dn)2Cje, carries a partial triplication of chromosome 16 which includes the Grik1 gene. This trisomic mouse mice could be a good model to understand the role of GluK1 in synaptic transmission and its putative influence in Down's phenotype. We will analyze the behaviour of synaptic responses in several places like the spinal cord and the necortex under stimulation of dorsal roots, thalamocortical projection and intracortical cell populations. We will relate these actions to phenotypic characteristics of Down's syndorme.
Methodology: Mice genotyping by qPCR. In vitro patch clamp recordings in acute brain slices.

WP.1.2. Supervisor: Dr. Santiago Canals
Project title:

Objectives: 1.
Methodology:

Student's name : Jego Pierrick / Laboratory of Dr Angel Barco, Regulation of Gene Expression and Synaptic Plasticity
Instituto de Neurosciencas (INA), Alicante, Spain

Origin : Born the 20 of December 1985 in Rennes (France)

Education : Master 1 of NeuroPhysiology in Rennes 1 university (2006), Master 2 of Marine Biology in Nantes (2007), Master 1 and 2 of Neurosciences in Victor Segalen (Bordeaux II) university (2008-2009)

Project title : epigenetics, the role of the histone demethylase KDM5C in learning and memory. It has been reported that the mutation of this enzyme situated on the X chromosome leads to a mental retardation (X linked mental retardation diseases). So I am studying the effects of an overexpression and a repression of this enzyme on mice's behaviour, including cognition, learning and memory, locomotion

WP.1.3. Supervisor: Dr. Maurice Garret
Project title: Characterisation of GABAergic neurotransmission within the globus pallidus and subthalamic nucleus network in a rat model of Parkinson’s disease.

Objectives: It is well established that neuronal activity within networks rely on the functional properties the receptors, their number, and their localisation. Moreover, alterations in GABAA receptor function contribute to the symptoms of many neurological diseases and psychiatric disorders. The basal ganglia are a group of subcortical nuclei involved in motor control. Within the basal ganglia, the Globus pallidus through is widespread GABAergic projections controls the activity of the entire network. Activity pattern of Globus pallidus neurons is profoundly altered in Parkinson’s disease, which contribute to the development of pathological oscillations in the basal ganglia. The PhD project aims to understand synaptic plasticity of pallidal GABAergic inputs by combining electrophysiology with fluorescent immunohistochemistry, in order to define changes in GABAA receptors functions and GABAergic network remodelling, in a rodent model of Parkinson’s disease.

Methodology: immunocytochemistry ; Correlation between synaptic function and structure. Development of image analysis software to quantify synaptic contacts in the brain.
Secondment : Explora Nova

Student's name : Zhuowei Du

WP.1.4. Supervisor: Dr. Jack Mellor
Project title: The role of cholinergic receptors in synaptic plasticity.

Objectives: The neurotransmitter acetylcholine is critical for memory formation in the brain. This is proposed to occur via activation of muscarinic receptors leading to an enhancement of NMDA receptor (NMDAR) function which then facilitates the induction of synaptic plasticity. We have recently discovered a link between muscarinic receptor activation and NMDAR facilitation involving SK channels but the precise SK channel subtypes involved and the mechanism by which they are modulated are not known. In this project we intend to collaborate extensively with Neurosearch who have a wealth of knowledge on the function and pharmacology of SK channels to investigate the specific SK channel subunits involved, how they are modulated and how they enhance NMDAR function.
Methodology: Electrophysiological techniques in hippocampal slices coupled with either pharmacological or genetic knockout or knockdown techniques to remove or block specific ion channels or components of intracellular signalling pathways. High-resolution calcium imaging using multi-photon microscopy. Computational modelling of synaptic conductance and calcium concentration.

Student's name: Valeria Olivo / Lab: Jack Mellor's lab, University of Bristol

Education: BSc in Biotechnologies of Health at University in 2008 and MSc in Pharmaceutical Biotechnologies in 2009, both at University of Naples "Federico II"

Origin: Naples, Italy

Project title : the project is called "The role of cholinergic receptors in synaptic plasticity" and it aims at understanding how muscarinic receptors can enhance NMDA receptor function and facilitate synaptic plasticity. As SK channels are shown to be involved in this process, the goal is to identify which subunits are involved, how these channels are modulated and how they enhance NMDAR function through electrophysiological techniques.

WP.1.5. Supervisors: Pr. Valentin Nägerl and Dr. Giovanni Marsicano
Project title: Role of the endocannabinoid system in morphological plasticity.

Objectives: The proposed project is aimed at investigating the role of the endocannabinoid system (ECS) in structural plasticity of hippocampal synapses. The cannabinoid receptor type 1 (CB1) is the most abundant G protein coupled receptor in the brain and this may explain the diverse involvement of the ECS in neuronal functions. This project will investigate the relationship between ECS activation and rapid morphological plasticity of neurons in physiological and pathophysiological conditions. The use of advanced genetic mouse models and high resolution time lapse microscopy techniques (STED) in defined experimental settings will allow identifying the cross-talk between the ECS and actin dynamics in the regulation of structural plasticity of synaptic connections.

Methodology: STED microscopy in living cells to detect in real time actin dynamics (use of fluorescent Lifeact expression, specifically binding to polymerized actin), hippocampal neuronal cultures (dissociated and organotypic), llectrophysiological stimulation of synaptic activity, excitotoxic protocols, ex vivo morphological studies from behaving mice.

Student's name: Philipp Bethge / Synaptic Plasticity and Superresolution Microscopy / Endocannabinoids and Neuroadaptation - IINS Bordeaux

Education : BSc Psychology Landau Germany, MSc Neuroscience Magdeburg Germany

Origin : German

Project: Endocannabinoids (2-Arachidonoylglycerol and Anandamide) are retrograde messengers that mediate both short term synaptic plasticity such as depolarisation induced suppression of inhibition (DSI) (Wilson and Nicoll 2001) as well as long term depression of inhibition (iLTD)(Chevaleyre and Castillo 2003). For decades, synaptic plasticity has been studied focusing on excitatory synapses and a structure-function relationship, such as the highly positive correlation of synaptic glutamate sensitivity with spine volume, has been developed(Matsuzaki, Ellis-Davies et al. 2001). Although the activity state of the hippocampus is tightly regulated by a balance between excitatory and inhibitory neurotransmission, surprisingly little is known if the structure-function relationship that governs excitatory neurotransmission holds true for inhibitory synapses as well. In this project, I will like to take advantage of two-photon microscopy and STED imaging to investigate the structural implications of endocannabinoid signaling in short- as well as long term plasticity paradigms, with a special emphasis on fast processes.

WP. 2.1. Supervisor: Dr. Dominique Muller
Project title: Regulation by mental retardation protein PAK3 of PSD formation in newly formed synapses

Objectives: Expression of mutant PAK3 responsible for mental retardation in humans leads to defects in the morphology and function of excitatory synapses, with in particular an increased number of thin, immature spines. The PhD project will consist in testing this hypothesis and identify the role of PAK3 in regulating dynamics of PSD proteins expressed at the synapse, particularly in newly formed synapses. the expression of PSD proteins. Experiments will specifically focus on PSD-95, GluR1/2 receptors and N-cadherin to determine the sequence and exact timing of expression of these proteins under control conditions, following expression of mutant PAK3, constitutively active forms of the protein or interfering peptides. Additionally, experiments using FRAP or proteins tagged with photoactivatable EGFP will be carried out to determine the kinetics of expression of these proteins under the different conditions tested. Finally, another part of the project will be to examine the role of synaptic activity on the dynamics and kinetics of expression of these proteins.

Methodology: the project will be based on the use of organotypic hippocampal slice cultures and gene-gun mediated transfections; analyses will be performed using repetitive confocal imaging, FRAP and photoactivatable EGFP.

Student's name : Katalin Sari / Laboratory: Dominique Muller lab since October 2010 CMU, University of Geneva

Education
Laboratory animal science and experimental design, Hungary, 2009
M.Sc. in Biology, Hungary, 2007

Origin : Hungary

Project title: Regulation by mental retardation protein PAK3 of PSD proteins
Organotypic hippocampal slice cultures were used for investigating the dynamic regulation of PSD proteins by mental retardation protein PAK3. Neurons were co-transfected using a PSD95 fused with photoactivatable EGFP plasmid and DsRed2 plasmid. The photoactivation was carry out by 2 photon laser microscopy, and the level of fluorescence associated with expression of PSD-95-EGFP monitored by single photon laser-scanning. With this approach I investigate the role of the mental retardation protein PAK3 on the dynamics and kinetics of expression of PSD95 protein. To do this I compare the kinetics of PSD-95-EGFP before and after application of the p21-Activated Kinase Inhibitor III, IPA-3. The preliminary data suggest that the kinetics of PSD-95-EGFP is not very much modified under control conditions by the PAK3 inhibitor. The next step will be to examine whether PAK3 inhibition affects the trafficking of PSD-95 following spine activation, using a 2-photon uncaging protocol to activate spines. A point mutation of PAK3 has been linked to nonsyndromic X-linked mental retardation. To investigate the kinetics of the PSD molecules will enable us to understand better the molecular basis of the nonsyndromic X-linked mental retardation.

WP. 2.2. Supervisor: Dr. Maria Passafaro
Project title: Characterization of genes involved in mental retardation.

Objectives: Human Mental Retardation (MR) is a common and highly heterogeneous paediatric disorder with a very severe social impact. Genetic defects account for a large, but not well-defined, portion of all MR forms, ranging from 25 to 50%. The scientific work and knowledge concerning the role of proteins encoded by these genes lead to the hypothesis that MR is a synaptopathy-like disorder: this is due in part to the fact that most of these proteins localize to pre- and/or post-synaptic neuronal terminals. Thus major goals of the PhD project will be i) analysis of the function of several different genes carrying mutations causing MR during neuronal development in vitro; ii) identification and characterization of protein complexes associated to molecules involved in MR.

Methodology: Primary neuronal cultures, bran slices culture, yeast two hybrid screening, RNA interference and overexpression of mutant using lentivirus vectors. In vitro patch clamp and extracellular recordings

Student's name : Jonathan Zapata / Laboratory Dr Maria Passafaro, Consiglio Nazionale delle Ricerche, Milano

Education : – Master's degree in Neurosciences and Neuropsychopharmacology, university of Bordeaux
II. – Two-years technical degree in biological analyzes, Bordeaux.
Origin : University of Bordeaux II

Project : Mental retardation (MR) is defined as a global deficiency in cognitive function (IQ<70) with an onset during childhood and associated with a diminished ability to adapt to the daily demands of a normal social environment. Over 10% of all cases of MR are thought to be X-chromosome linked (X-LMR). Mutations in various regions of one of the shroom family proteins, shroom4 encoded by KIAA1202 gene have been associated with cases of X-LMR. The Shroom family is a small family of actin binding proteins that are able to induce cytoskeletal changes and thereby influence cellular function and morphology. My present study aimed to characterize the shroom4 protein in the brain, in particular to investigate the possible role of shroom4 at the synapse

Methodology : Primary neuronal cultures, bran slices culture, yeast two hybrid screening, RNA interference and overexpression of mutant using lentivirus vectors. In vitro patch clamp and extracellular recordings. In-vivo imaging.

WP. 2.3. Supervisor: Dr. Nathalie Sans
Project title: Functional characterization of a Scribble1-Vangl2-Lgl1 complex in the hippocampal network of new animal models of mental retardation

Objectives: Many proteins containing PDZ domains, associate to glutamate receptors (GluRs) and are required for protein (eg GluRs) trafficking and anchoring to synapses. Recently, we have identified Scribble1, a PDZ domain protein, as a new binding partner for GluRs, and as a potential regulator of their trafficking. We aim at unravelling novel regulatory mechanism implicating the PDZ domains of Scribble1 and maybe different active or inhibitory complexes. We have identified interacting partners of Scribble1. The interactions and the molecular cascades set up by these proteins are primordial for synaptogenesis, and synaptic plasticity and we wish to analyze these different complexes and their role in the trafficking of receptors. The determination of the specificity or the strength of the interaction most probably involves allosteric interactions. For this project, we will use different biochemical assays to validate each interaction in mammalian cells, and assess the role of each complex in the regulation of receptor trafficking. We will also analyze the core part of these assemblies by X-ray crystallographic analyses. Finally, we will check whether some C-terminal PDZ-binding motif could disrupt the assembly of complexes, thereby impacting on cell-surface trafficking and anchoring properties.

Methodology: Biochemistry to charaterize synaptic complexes, cell biology analysis of the different complexes, X-ray crystallographic analyses.
Secondment : Bio-Xtal

Student's name : Vera Pinheiro / Laboratory: Dr. Nathalie Sans - INSERM U862, Bordeaux

Education and training:
Molecular and Cell Biology Degree - Faculdade de Ciências e Tecnologia - Universidade Nova de Lisboa (FCT/UNL), Portugal
Cell and Molecular Biology M.Sc. - Faculdade de Ciências e Tecnologia - Universidade de Coimbra (FCT/UC), Portugal
Thesis Project as an Erasmus student at the Dept. of Biology - Università di Trieste, Italy: Regulation of Brain Derived Neurotrophic Factor mRNA translation in chronic epilepsy.

Title (provisory): Characterization of Scrib1 protein interactions in synapses and their importance for autism spectrum disorders
Scribble has been previously described as a key regulator of apicobasal polarity in Drosophila; its mammalian homolog, Scrib1, is known to be implicated in cancer, neural tube closure, and planar cell polarity (PCP). Scrib1 protein is composed by multiple interacting domains, namely 16 leucine-rich repeats (LRRs) in the N-terminal and 4 PDZ (PSD-95/Discs-large/ZO-1) domains in the C-terminal and is present in dendritic spines. Many of the organizing scaffold proteins in the PSD, implicated in trafficking and anchoring of synaptic proteins, contain PDZ domains, suggesting an additional role for Scrib1 at the postsynaptic level.
To further characterize the role of Scrib1 in both developing and adult nervous system, our lab is been using circletail mutants (crc), a mouse model for PCP defects, in which both PDZ 3 and 4 of Scrib1 protein are absent. Particularly in the hippocampus, these mutants are characterized by having an increased number of enlarged spines and post-synaptic density (PSD). This can be the result of the observed mislocalization of the signaling pathway downstream of Scrib1 and consequent overall activation of Rac1, a known regulator of actin dynamics. Moreover, Scrib1-deficient mice exhibit features related to autism spectrum disorders, namely enhanced learning and memory abilities and impaired social behavior.

Therefore, the main goal of this project is to discover which specific interactions are responsible for the phenotype of crc mutants. First, we're going to analyze protein-protein interactions through yeast two-hybrid, using PDZ3 and PDZ4 as baits and a mouse brain cDNA library as a prey. The resulting interactions will then be validated by biochemical assays, such as co-immunoprecipitation, and further characterized in primary hippocampal cultures. Finally, in collaboration with BioXtal, a structural biology company, we will determine the structure of these putative protein-protein interactions and develop a specific blocking peptide to efficiently disrupt the interactions.

WP. 3.1. Supervisor: Dr. John Hanley
Project title: Intracellular signaling mechanisms during ischaemia

Objectives: Brain ischaemia involves changes in the subunit composition of synaptic AMPARs, and may involve similar mechanisms to the well-studied phenomena of LTD and LTP. The project will investigate changes in small GTPase activation during ischaemia and study the signaling events that bring about these changes following insult, and the effect on AMPAR trafficking. Preliminary data from our lab suggest that Arf1 signaling is important in LTD, with possible roles for Rac and Cdc42. This will be novel, exciting research that will contribute to our knowledge of the mechanisms of brain ischaemia.

Methodology: In vivo rodent models of stroke will provide brain tissue for biochemical experiments to assess GTP-bound (activated) levels of GTPases in stroke vs control conditions. This will be backed up with in vitro models (Oxygen/Glucose Deprivation of hippocampal cultures). Mutants of GTPases will be expressed (and/or endogenous levels knocked down) in cultured neurons, and the effect on OGD-induced AMPAR trafficking determined by a combination of fixed and live cell imaging.

Student's Name: Elena Blanco Suarez /Laboratory: John Hanley. Department of Biochemistry, University of Bristol (UK).

Education: Biology Lic. in Universidad de Oviedo. Master in Biochemistry, Molecular Biology and Biomedicine in Universidad Complutense de Madrid.

Origin: Asturias, Spain.

Project:GTPases and AMPAR trafficking during oxygen/glucose deprivation in hippocampal and cortical neurons
Oxygen-glucose deprivation (OGD) mimics the conditions produced during brain ischemia. OGD induces a change in AMPAR subunit composition at hippocampal synapses, resulting in the expression of Ca2+-permeable AMPAR which contribute to excitotoxicity and therefore, neuronal death. Cortical neurons are less vulnerable to ischemia than hippocampal neurons, but the mechanisms behind this differential vulnerability are unknown. Small GTPases influence dendritic spine morphology and AMPAR trafficking, but their role in ischemia is unclear.
I am using surface biotinylation and imaging to investigate AMPAR trafficking during ischemia in hippocampal and cortical cells. In addition, I am investigating how GTPase activation changes in response to ischemia, with the aim of finding out how GAPs and GEFs might be involved in the possible regulation and protection against ischemic insult.

WP. 3.2. Supervisor: Dr. Jeremy Henley
Project title: Mechanisms and neuroprotective properties of SUMOylation of synaptic protein in models of ischaemia.

Objectives: Massive amounts of protein SUMOylation occur when neurones are subjected to stressors and it has been proposed that SUMO conjugation may be neuroprotective mechanism. Some of the proteins that undergo SUMOylation are synaptic but the molecular mechanisms that mediate neuroprotective events and the precise identities of the proteins that are SUMOylated remain to be determined. The aim of this project is to define specific synaptic SUMO target proteins and elucidate their roles in neuroprotection.

Methodology: OGD of dispersed rat hippocampal neuronal cultures and acute or organotypic cultured rat hippocampal slices as model systems to study ischaemia. Expressed of Sindbis virus SUMO-1, SUMO-2/3 and SENP-1 on neuronal survival following OGD using wild-type SUMO and SENP. The viability in OGD and non-OGD cells will be evaluated by quantitative analysis of virally transduced (GFP-containing) and non-transduced neurones, stained with markers such as propidium iodide and Hoechst, as well as by measuring LDH release. In all cases Western blots and immunocytochemistry to determine the substrates and extent of protein SUMOylation.

Student's name : Fernando Josa Prado / Laboratory: Prof. Jeremy Henley's lab. Dept. Biochemistry. University of Bristol

Education:
Biology Degree at the Universidad Autónoma de Madrid in 2008.
D.E.A in Genetics and cellular Biology (Diplome of Advanced Studies, granting the researcher's sufficiency) at the CMBSO/Universidad Autónoma de Madrid in 2010.
Expert in psychotherapy and Ericksonian Hypnosis at the Erickson Institute in Madrid, 2010.

Origin: Madrid, Spain

Project: The project consists on assessing the role of PIAS3 (Protein Inhibitor of activated STAT3) in neurons. PIAS3 is a SUMO E3 Ligase, and its basic characterization in this type of cell is still unclear. This work will involve the mentioned characterization as well as finding more concrete roles for PIAS3 in neuronal molecular pathways and mechanisms.

WP. 3.3. Supervisor: Dr. Oliver Schlüter
Project title: Analysis of the mechanisms of signaling scaffolds in synaptic plasticity

Objectives: Determine the role of alternative N-termini of PSD-93 on synaptic transmission. Decipher the role of individual domains of DLG-MAGUKs on synaptic function. Measure changes in the synaptic composition and post-translational modifications upon perturbations of DLG-MAGUK signaling complexes. Identify the role of signaling scaffolds in rodent learning paradigms.

Methodology: Viral mediated genetic manipulation, whole cell patch clamp electrophysiology, proteomic analysis of synaptic changes in phosphorylation status and protein content, behavioral assays with conditional DLG-MAGUK KO mice

Student's name : Tanmoy Samaddar / Lab: Molecular Neurobiology group of the Dr. Oliver Schlüter, European Neuroscience Institute, Göttingen

Education: Bachelor of Science in Microbiology at the University of Calcutta, Kolkata, India
Masters in Science in Biotechnology, Department of Biotechnology, Univ. of Calcutta, Kolkata, India
Publication-Tripathi R, Samadder T, Gupta S, Surolia A and Shaha C* (2011) Anti-cancer activity of a combination of cisplatin and fisetin in embryonal carcinoma cells and xenograft tumors. Mol Cancer Thermolcanther.0606.2010; Published OnlineFirst January 7, 2011; doi:10.1158/1535-7163.MCT-10-0606

Origin: Kolkata, India.

Project details: Synapses of mammalian central nervous system are specialized structures designed for rapid and efficient transmission of signals between neurons. Our lab mainly focuses on the molecular basis of learning and memory in the post-synaptic part of neural networks and the crucial role of Dlg-MAGUK proteins, one of the major components of the post-synaptic density. Using newly developed techniques like molecular replacement, mouse genetics and viral mediated gene manipulation in cultured neurons as well as in-vivo experiments, the functions of the MAGUKs are aimed to be dissected. My project in particular would aim at building up a post-synaptic protein interactome of the Dlg-MAGUKs using SILAC and Mass spectrometry methods. Knowledge gained from this study will help to divulge the highly dynamic interactions at the post-synaptic density and the spatio-temporal changes during activity dependent modulation.

WP. 3.4. Supervisor: Dr. Monica DiLuca
Project title: Characterization of DA and Glu interaction in experimental parkinsonism

Objectives: the involvement of glutamatergic and dopaminergic systems appears to be the peculiar feature of striatal plasticity and its disruption might underlie the pathogenesis of basal ganglia neurodegenerative diseases including PD. Indeed The induction of synaptic plasticity in the striatum requires interaction between dopamine (DA) and glutamate receptors; thus major goals of the PhD project will be i) the molecular analysis of postsynaptic glutamate receptors and/or subsynaptic cytomatrix-scaffolding proteins and their interactions with the DA system in physiological and pathological conditions making use of experimental model of parkinson disease (PD), ii) analysis of physiological and pathological protein-protein interactions and characterization of critical molecular targets involved in PD.

Methodology: primary neuronal culture, brain slices culture, overexpression of mutant using lentivirus vectors, cell permeable peptides as tool to manipulate synaptic protein complexes.

Student's name : Jennifer Stanic / Lab: Monica Di Luca, Department of Pharmacological Sciences, University of Milan

Education: 2010 Master II (Master degree) Cellular Biology, Physiology & Pathology, Neurobiology specialty – Paris Descartes University Magma Cum laude
2009 Master I Biology-oriented Pharmacology & Toxicology – Paris Descartes University Cum laude
2008 Bachelor degree of Life Sciences, Biology specialty – Paris Descartes University Cum laude

Origin: Paris, France

Project: My project involves the characterization of DA and Glu interaction in experimental parkinsonism by studying a protein called Rabphilin 3A that was identified as possibly interacting with NMDA receptor NR2A subunit by a two- hybrid experiment. Rabphilin 3A (Rph3A) is a synaptic vesicle-associated protein that was first identified as a binding partner of the GTP-bound Rab3A, a member of the Rab family of GTPases implicated in vesicle docking/fusion reactions. Moreover, different studies indicated that Rph3A can regulate exo- and endocytosis processes. Its has also been shown to be modulated in models of Huntington disease and in the alpha-synuclein model of Parkinson disease. The aim of the project is to characterize Rabphilin in the Post-synaptic density, to describe its interaction with NR2A subunit and its function in the post synaptic density and potential role in the localization of the NR2A subunit at the synapse in physiological conditions as well as in Parkinson disease.

WP. 3.5. Supervisor: Dr. Carlo Sala
Project title: Molecular mechanisms regulating postsynaptic density protein assembly and plasticity

Objectives: Synapses are the sites of most information processing in the brain, and changes in their molecular and functional composition are likely critical for the modulation of synaptic efficacy in normal and pathological conditions. Neuronal activity causes long lasting changes in synaptic structure and function by regulating dendritic synthesis of proteins functional associated to synapses. Recent evidence suggests that neuronal activity modulates eukaryotic elongation factor 2 (eEF2) pathway in dendrites, thus yielding activity dependent synthesis of proteins known to be involved in the regulation of synapse plasticity. The major goal of the PhD project will be: i) to characterize the molecular mechanisms regulating activity dependent translation of synaptic proteins in dendrites ii) to determine the molecular mechanism of synaptic dependent regulation of eEF2 kinase (eEF2K) activity on eEF2 or on other potential substrate.

Methodology: primary neuronal culture, brain slices culture, overexpression of mutant using lentivirus vectors, cellular imaging, biochemical and proteomic techniques, development of comparative proteomic approaches.

Student's name : Christopher Heise / Laboratory:Prof. Carlo Sala at CNR, Institute for Neuroscience, Milano

Origin: Born in Hameln (Lower Saxony, Germany)

Education:Main study in neuroscience at the Otto-von-Guericke-University Magdeburg.

Project title :In my PhD thesis I am trying to elucidate new functions of Elongation factor 2 kinase (eEF2K) associated with synaptic plasticity. For this, common tools of molecular biology and biochemistry are implemented in various experimental models. However, also tools are used which are less common in neuroscience such as stable isotope labeling by amino acids in cell culture (SILAC).

WP. 3.6. Supervisor: Dr. Stefan Eimer
Project title: Regulation of nicotinic receptor trafficking and synaptic function in C. elegans.

Objectives: The PhD project aims to precisely characterize the molecular mechanisms how UNC-50 and UNC-74 affect nAChR trafficking. At steady state, UNC-74 localizes to the endoplamic reticulon while UNC-50 is found mainly at the Golgi apparatus. This suggests that both molecules may act sequentially for nicotinic receptor transport through the secretory system. The PhD project aims at finding new molecules that are required for cell surface expression and function of nicotinic acetylcholine receptors in a subtype specific manner, at characterizing the mechanisms regulating nicotinic receptor trafficking. The live cell imaging will be complemented by ultrastructural studies using high pressure freeze (HPF) electron microscopy (EM). To quantify and correlate protein distributions with cellular ultrastructures and morphology at a nanometer resolution we have developed protocols that combine HPF sample preparation with immunofluorescence staining methods allowing correlative microscopy.

Methodology: Biochemistry, Cell Biology, Genetics in C. elegans, live cell Imaging, Electrophysiology, high pressure freeze Electron Microscopy.

Student's name : Suekyoung Jeon Lab: Molecular Neurogenetics of European Neuroscience Institute, Göttingen, Germany, Prof. Stefan Eimer

Education: M.Sc., Microbiology, Konkuk University, South Korea; B.Sc., Molecular Biology, Konkuk University, South Korea

Origin: Seoul, South Korea

Project: Topics_Studying the regulation of nicotinic receptor sorting and transport in C. elegans.

Purpose of the PhD project is to work out the molecular details how nAChR are sorted by UNC-50 and what are the molecular determinants. The Project will further use the oocyte expression system to study nAChR transport in a heterologous system that allows us to characterize them by electrophysiologie. In addition, we have generated tagged UNC-50 proteins and we will try to purify UNC-50 binding partners by using biochemical approaches in combination with mass spectrometry. The biochemical approaches will be complemented by a genetic screen for unc-50 suppressors. On the other hand the project will rely heavily on high resolution imaging techniques as we are trying to follow nAChR trafficking live through the intracellular systems by using point scanning and spinning disc confocal imaging in combination with STED and electron microscopy.

WP. 3.7. Supervisor: Dr. Daniel Choquet and Dr. Jean Baptiste Sibarita
Project title: Development of a new technique for fast multiphoton super-resolution imaging of thick biological samples

Objectives: The PhD project aims to develop an innovative fast multiphoton imaging technique which will allow imaging of thick biological samples at the single molecule level. This technique will be applied to study receptor trafficking in 3D in brain slices and to associate this with the analysis of receptor trafficking in vivo. The setup will be designed to be versatile in order to implement, on the same microscope, multiphoton SPT-PALM (Single Particle Tracking & PhotoActivation Localization Microscopy) and SIM (Structured Illumination Microscopy) techniques in order to reach a sub-wavelength resolution.

Methodology: Multi-photon microscopy, video-microscopy, Femto-second laser fiber coupling, image processing and analysis.
Secondment : Amplitude systèmes.

Studen's name : Kalina Hass

WP. 4.1. Supervisor: Dr. Miguel Maravall
Project title: Short-term plasticity and regulation of synaptic responses by activity under natural conditions

Objectives: A key functional property of synapses is their short-term plasticity, through which the magnitude of a synaptic connection is dynamically regulated in activity-dependent manner. We have recently found that thalamocortical synapses in the rodent barrel cortex, which are considered to be canonical examples of synapses that undergo short-term depression, display strikingly heterogeneous short-term dynamics when stimulated with irregular or natural stimulus trains. While all thalamocortical synapses depress under regular stimulus trains, irregular trains elicit responses with a fast facilitating component that varies widely across different synapses. This facilitating component and its heterogeneous expression have profound implications for the codes used by thalamocortical populations of neurons to represent sensory information.
In this project we will examine two crucial aspects of this heterogeneous thalamocortical short-term plasticity. First, regulation of synaptic efficacy can occur through several mechanisms, but their joint action under irregular or natural stimulation conditions remains poorly understood. We will analyze the specific contributions of kainate receptor-mediated regulation (kainate receptors are known to be presynaptically expressed in thalamocortical synapses) and of disynaptic inhibition (inhibition in the thalamocortical pathway contributes strongly to overall synaptic drive and is also dynamically regulated). Second, thalamocortical flow of sensory information in the rodent whisker system occurs through several parallel pathways. Short-term dynamics could be regulated on a pathway-specific basis or even on a pre- or postsynaptic cell-specific basis. We will test whether this is the case by recording the responses of selectively activated synapses.

Methodology: In vitro patch clamp recordings from thalamorecipient neurons in different layers of mouse barrel cortex. Synaptic activation via extracellular electrical stimulation as well as optogenetic tools (selective expression of fast channelrhodopsin variants together with optical stimulation).

Student's name : Giovanni Ferrat / iReaserch Group: Dynamics and plasticity of cortical sensory responses
Principal Investigator: Miguel Maravall Universidad Miguel Hernández Consejo Superior de Investigaciones Científicas
Instituto de Neurociencias, Campus de San Juan | Sant Joan d'Alacant
Alicante | España

Origin: Florence, Italy

Education: Bachelor Degree in Biotechnology, Departments of Experimental Pathology and Oncology, University of Florence, Italy .Master Degree in Neurobiology, Department of Animal and Human Biology, University of Turin, Italy

PhD Project: "Short-term plasticity and regulation of thalamocortical synaptic responses in the whisker system"
Context dependence, a key attribute of information processing in higher brain areas, is mediated by activity-dependent regulation of synaptic efficacy. Several mechanisms, from kainate receptor activation to calcium buffering, underlie this regulation, but their action remains poorly understood in many specific synapses. We have recently discovered that thalamocortical synapses in the mouse barrel cortex have highly heterogeneous dynamic behaviour, which could greatly affect communication of sensory information to the cortex.
To gain insight into the mechanisms underlying the diverse behaviour of thalamocortical synapses in the barrel cortex, we will first analyze synaptic responses to naturalistic temporal patterns of stimulation and will isolate the roles of kainate receptors and of inhibition in regulating the behaviour. Second, thalamocortical flow of sensory information in the rodent whisker system occurs through several parallel pathways. Short-term dynamics could be regulated on a pathway-specific basis or even on a pre- or postsynaptic cell-specific basis. We will test whether this is the case by recording the responses of selectively activated synapses.

WP. 4.2. Supervisor: Dr. Christophe Mulle
Project title: Analysis of synaptic plasticity in CA3 pyramidal cells using optogenetics

Objectives: CA3 pyramidal cells and their afferent mossy fibers originating from the dentate gyrus play an important role in the encoding of novel information. Mossy fiber synapses display unique forms of pre- and postsynaptic plasticity. The objective of the PhD project is gain insight into the mechanisms and the physiogical consequences of synaptic plasticity in the intact mouse brain. For this we aim at combining electrophysiological recordings in vivo with optogenetic activation of excitatory dentate gyrus cell populations expressing channelrhodopsins. Optogenetic methods will first be validated in the in vitro slice preparation. We will then compare selected mutant mice in which in vitro synaptic plasticity is affected.

Methodology: In vitro patch clamp recordings. In vivo combined intracellular and extracellular recordings. Activity manipulation via optogenetic tools (channel rhodopsin expression in the dentate gyrus). Stereotaxic viral infection for the expression of Channel rhodopsins.
Secondment : Neurosearch.

Student's name : Stefano Zucca

WP. 4.3. Supervisor: Dr. Marc Landry and Dr. Alexandre Favereaux
Project title: Regulation of the synaptic plasticity of spinal cord neurons by microRNAs in pain condition.

Objectives: In pain conditions, spinal cord neuronal networks undergo many changes, both structural and functional, that are substrates for the persistence of pain. This neuronal plasticity is the result of multiple molecular mechanisms among which is the regulation of the proteins implicated in synapse formation. The purpose of this thesis is to investigate the control exercised by microRNAs (miRNAs) on the synaptic proteins translation. The first part of the work will be to identify changes in the microARNs and the mRNA expression repertoires in an animal model for pain. Then, with bioinformatics tools, the ESR will try to connect these two patterns of expression considering a coordinated inhibition of multiple synaptic proteins (synapse scaffolding proteins, ionic channels, receptors …) by a sub-population of miRNAs. In a second time, the ESR will artificially modulate the expression level of these miRNAs in culture cells to measure their impact on synapses.

Methodology: photonic and electronic imaging techniques, calcium imaging, electrophysiology, animal model for pain.
Secondment : Explora Nova.

Student's name: Sara Elramah / Lab: CNRS, Interdisciplinary Institute for Neuroscience (IINS), Dr. Landry Team.

Education: Bachelor of Science - Faculty of Sciences, University of Khartoum, Sudan.
Master - Faculty of Sciences, Alexandria University, Egypt.

Origin: Port Sudan, Sudan.

Project title : MicroRNAs (miRNAs) are classes of small non-coding RNAs, which function as negative regulators of target mRNA expression at the posttranscriptional level. Because the long-lasting changes in cancer pain sensitivity are accompanied by altered gene regulation, miRNAs expressed in nociceptive pathways may influence the development and maintenance of pain. Our interest is to detect the difference in genes' expression and their regulation by miRNA, in bone cancer pain condition. Our strategy includes; developing of bone cancer model, analysis of changes in the miRNAs and mRNA expression, confirming the differences in the expression of these candidates in vitro and finally testing their effect in vivo using classical behavioural tests. Our goal is to identifying new miRNA mechanisms involved in cancer pain and their potential as therapeutic targets.

WP. 4.4. Supervisor: Dr. Zafar Bashir
Project title: Assessing changes in synaptic plasticity in perirhinal cortex.

Objectives: Synaptic plasticity (LTD) has been to shown to be crucial for visual recognition memory. The ESR will examine changes in LTD in perirhinal cortex in models of schizophrenia. It is envisaged that students may use techniques such as fast cyclic voltammetry to assess changes in neurotransmitter release or assessing behavioural/cognitive deficits associated with schizophrenia. They will spend part of the time in Bristol using a range of electrophysiological techniques to investigate any effects on synaptic plasticity and biochemical and imaging techniques to address the underlying causes of changes in synaptic plasticity.
Methodology: fast cyclic voltammetry; behavioural/cognitive assessment; in vitro electrophysiological techniques; biochemical and imaging techniques

Student's name : Medhi Bhouri

WP. 4.5. Supervisor: Dr. Christian Lüscher
Project title: Channel Rhodopsin 2 as a tool to activate midbrain dopamine neurons

Objectives: Engineer a transgenic mouse where ChR2 is selectively expressed in DA neurons of the midbrain.

Methodology: Use of DAT-Cre transgenic mouse line in combination with AAV viral vectors, stereotaxic injections, blue light stimulation and ex vivo electrophysiology. Measurement of DA levels using carbon fibers to demonstrate efficacy
Secondment : Bioxtal

Student's name : Tifei YUAN /Lab: Christian Luscher

Education: M.phil University of Hong Kong
B.Sc. 2007 Sun Yat-sen University

Origin: China

Project title : The changes of NMDARs in ventral tegmental area dopamine neurons of mice following acute cocaine exposure.The main technique is whole-cell patch clamp on acute slices of mice brain. The final goal of the study is to understand the neural plasticity changes in the context of addiction.

WP. 4.6. ESR21 at UniGe (36 months). Supervisor: Christian Lüscher
Project title: Target-cell type specific facilitation and depression in cortical circuits.

Objectives:

Methodology:

Student's name :Sebastiano Bariselli/

Origin: Italy

Project :

WP. 4.7. Supervisor: Dr. Anthony Holtmaat
Project title: Imaging of activity-related structural changes in synapses in vivo.

Objectives: Structural neuronal changes such synapse formation and elimination as well as changes in synapse size could underlie long-term storage of memory or adaptations to new experiences. Recently it has been shown that a change in sensory experience drives the formation of new synapses (in the order of 10%) on L5 pyramdial cells in barrel and visual cortex. After peripheral neuronal lesions even the entire complement of synapses is replaced in the areas that functionally change. Such micro-structural changes could be a potential substrate to functionally compensate for loss of neuronal function after local cortical damage and in neurodegenerative disorders. We aim to investigate structural plasticity after local brain damage and explore possibilities to promote structural plasticity in these paradigms by sensory or optical stimulation of neurons, using enriched environments and channel rhodopsin (ChR). We will use long term in vivo two-photon laser scanning microscopy to image dendritic spines in GFP/RFP expressing L2/3 pyramidal neurons in the neocortex. L2/3 pyramidal cells will be labelled using viral vectors or by in utero-electroporation.

Methodology: In vitro 2-photon laser scanning micrsocopy. Viral vector technology. In vivo intracellular and extracellular recordings. Intrinsic signal optical imaging.

Student's name : Vassilis Kehayas / Laboratory: Structural synaptic plasticity Lab, Prof. Anthony Holtmaat

Education: M.Sc. in the "Brain and Mind Sciences" (University of Crete), B.Sc. in Biology (University of Crete)

Origin: Greece

Project: The aim of our project is to explore activity-dependent structural plasticity of layer 2/3 neurons in the barrel cortex in vivo. Our methodological approach will employ the use of optogenetic tools and viral vector technology, extracellular recordings, as well as two-photon laser scanning microscopy and intrinsic optical imaging.

WP. 4.8. Supervisor: Dr. Andreas Frick
Project title: Defects in neocortical circuits underlying cognitive disorders.

Objectives: It is a generally held belief that long-term memories in the cortex are stored in the strength of synaptic connections between neurons. However, this view has been challenged by recent studies. We now know, for example that the excitability of dendrites is a plastic property of neurons that can be induced by activity, experience, or disease. We hypothesize that changes in the dendritic excitability of prefrontal cortex neurons contribute to the formation of conditioned-fear memory. We will investigate this hypothesis in pyramidal neurons of the medial prefrontal cortex.

Methodology: Dendritic excitability of pyramidal neurons will be gauged using two-photon laser scanning microscopy for calcium imaging and for localized photochemical release of caged glutamate in combination with electrophysiology (somatic and/or dendritic recordings) in acute medial prefrontal cortex slices.

Student's name :Maria Szlapczynska