Research Associate (Studying Insulin Protection During Pancreatitis)
University of Manchester - School of Medical Sciences
Manchester, United Kingdom
Overall Purpose of the Job:
Acute pancreatitis is a serious and sometimes fatal inflammatory disease whereby the pancreas digests itself. This is caused by bile acids and ethanol/fatty acid metabolites. Recently, impaired metabolism and cytotoxic calcium overload have been suggested to be the cardinal events that trigger the disease regardless of the cause. There is strong clinical evidence linking diabetes (both type-1 and type-2) and the severity of pancreatitis. Moreover, using experimental animal models, our preliminary studies show that pancreatitis is worse in type-1 diabetic mice and mice in which the insulin receptor has been specifically deleted in pancreatic acinar cells of adult mice (PACIRKO mice). Furthermore, we have shown that insulin directly protects acutely isolated pancreatic acinar cells from cellular injury induced by pancreatitis-inducing agents. This was due to a metabolic shift towards glycolysis, sufficient to maintain ATP to fuel the plasma membrane Ca2+ pumps (PMCA) and thus prevent cytotoxic Ca2+ overload, even in the face of impaired mitochondrial function. The over-arching aim of this project is to further test the hypothesis that insulin directly protects pancreatic acinar cells during pancreatitis. This will be directly and unambiguously tested in PACIRKO mice using cellular and in vivo models of pancreatitis, in which exogenous insulin is administered using a hyperinsulinaemic-euglycaemic clamp. In addition, the molecular mechanism for this insulin protection will be determined by investigating the signalling pathways and metabolic phenotype induced by insulin. The successful outcome of this project will further corroborate insulin, and related agents, as possible treatments for acute pancreatitis, regardless of the precise causative factor. It is hoped that this multidisciplinary approach from molecular mechanism to whole organism disease state will have major clinical implications for the treatment of acute pancreatitis.
Due to the circumstances of the role; you must be available to work outside normal working hours.
Key Responsibilities, Accountabilities or Duties:
You are required to familiarise themselves with the University’s Equality and Diversity policies and to actively support these wherever possible.
Main Duties include:
- work independently and collaboratively as part of a multidisciplinary team;
- develop and oversea new techniques required for the project;
- maintain and organize results and research outcome;
- analyse and interpret results;
- participate actively in lab and group meetings;
- present and discuss the project at national and international meetings;
- communicate with the project leaders, collaborators and members of the lab;
- manage, supervise and train research technician and research students.
Techniques involved include:
- dissecting mouse tissues and preparation and isolation of primary cell culture;
- mouse colony maintenance; husbandry, breeding and genotyping;
- processing mouse tissue for histology;
- mammalian cell culture;
- SDS-PAGE, immunoblotting and ELISA;
- immunoprecipitation and pull-down assays;
- protein phosphorylation assays;
- proteomic analysis and mass spectroscopy (collaborative training);
- DNA, RNA and protein extraction and preparation from tissues and cells;
- RT-PCR and qPCR;
- recombinant protein expression in mammalian cells;
- metabolomics and fluxomic profiling (collaborative training);
- epifluorescence and confocal imaging;
- in vivo techniques; experimental models of pancreatitis, hyperinsulinaemic euglycaemic clamp (collaborative training).
Essential Knowledge, Skills, Experience and Qualifications:
- hold a Ph.D. in a biological science subject (or expect to hold a PhD by the time of appointment);
- possess good analytical skills and the ability to analyse and interpret complex data;
- good publication track record or experience of writing scientific papers/reports;
- be highly motivated, committed to research and be able to work collaboratively as part of a multidisciplinary team as well as develop his/her own initiatives;
- possess good organizational skills and good working practice;
- have the interpersonal and communication skills to deal with other staff, supervisor and collaborators in a helpful and friendly manner;
- be flexible with respect to working time commensurate with the project;
- have experience and a good working knowledge of protein biochemistry, such as immunoblotting, immunoprecipitation/pull-down assays, protein phosphorylation and proteomic analyses.
Desirable Knowledge, Skills, Experience and Qualifications:
Previous experience with the following techniques would also be an advantage:
- Fluorescence microscopy/imaging;
- Animal handling, husbandry, genotyping, and in vivo techniques (e.g. surgery);
- Tissue DNA, RNA extraction/preparation, RT-PCR and Q-PCR;
- Protein biochemistry (immunoblotting, ELISA, immunoprecipitation/pull-down assays, protein phosphorylation and phosphoproteomic analyses);
- Histology, immunofluorescence, immunohistochemistry;
- Metabolomics and fluxomics analyses;
- Molecular cloning techniques.