Proteomics Approaches to Measuring Neurogenesis

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Proteomics Approaches to Measuring Neurogenesis

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ESR6:  Design and validation of inhibitors of NMDA receptor signalling complexes.

This position has been filled

Supervisor:   Peter James
Host institution:   Lund University

Description:
This project aims to identify new components of nNOS signalling pathways of relevance to MAPK regulation by applying RNAi and proteomic methods together with existing leads. The interactions between these components will be characterised and defined. Subsequently cell permeable peptides will be developed to disrupt these interactions to allow validation and the functional consequences of disruption, in collaboration with consortium partners. Please see Cao et al 2005 JCB PMID: 15631993; Florio et al 2009 B J Pharm PMID: 19732061; Doucet et al., 2012 Pharmacol Ther PMID: 22133842; Li et al. 2013 J Neurosci PMID: 23658158.

ESR7:  Protein biomarker pathway analysis.

This position has been filled

Supervisor:   Peter James
Host institution:   Lund University
Duration:  36 months

Description:
The candidate will identify membrane markers of neurogenic cells with the long term goals of preparing novel imaging tools and improved biological understanding. This will be done in the laboratory of Peter James in Lund (Sweden) with secondments in the groups of Eleanor Coffey in Turku (Finland) and in the phage antibody development company Immunovia in Lund (Sweden).

(i) The first aim is to identify membrane protein markers of neurogenesis in order to be able to raise antibodies to allow the identification of neurogenic cells. Neurogenesis discontinues in ageing brain and subtractive proteomics will be used to identify markers by comparing neurogenic-incompetent cells with neurogenic competent cells from the dentate gyrus and ventricular zones of mouse brain. Laser dissection and FACS sorting using the proposed PSA-NCAM marker will be employed to enrich for these cells after gross dissection and tissue disruption. HPLC-MS/MS analysis of plasma membrane enriched preparations will be carried out using low-specificity protease digests methods developed in the James laboratory and by glycoprotein affinity capture.

(ii) A second approach to identify membrane markers of neurogenesis will be to utilize transgenic mice where GFP is driven by the DCX promoter. Accurate FACS sorting will be used to isolate neurogenic cells from adult brain.

The applicant must have experience working with animals, preferably in neurophysiology. Knowledge of proteomics techniques, both protein and peptide separation will be of great value. The ITN requires that the candidate be flexible and keen to work in several environments that cover the specialist areas that make up the work package.
The candidate will learn advanced mass spectrometry using label-free and isotopic labeling techniques as well as glycol-affinity capture and be trained in bioinformatics for the analysis of pathways and protein function.

ESR8:  Protein interaction analysis

This position has been filled.

Supervisor:   Peter James
Host institution:   Lund University
Duration:  32 months

Description:
The candidate will identify the substrates of JNK that regulate anxiety and depressive behaviour using phospho-proteomics in order to delineate the novel druggable pathway(s) involved in anxiety and depression. This will be done in the laboratory of Peter James in Lund (Sweden) in close collaboration with the group of Eleanor Coffey in Turku (Finland).

The applicant should have experience working with cell culture. Knowledge of proteomics techniques, both protein and peptide separation will be of great value. The ITN requires that the candidate be flexible and keen to work in several environments that cover the specialist areas that make up the work package.

The candidate will learn advanced mass spectrometry using label-free and isotopic labeling techniques as well as crosslinking interaction capture and be trained in bioinformatics for the analysis of pathways and protein function.