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Organelle dynamics in pancreatic β-cell physiology and pathophysiology and the importance of a membrane tethering protein in these dynamics
Sweden (IFMSA-Sweden) - Uppsala Universtitet, Uppsala
Department of Medical Cell Biology
Swedish and English (required)
Type of Research Project
- Basic science
What is the background of the project?
Direct physical contacts between cellular organelles constitute reaction centers in cells where rapid and specific transfer of ions, lipids and metabolites occur. Despite substantial lack of information about the composition of organelle contacts, it has become clear that the function of multiple organelles, including the endoplasmic reticulum (ER) and the mitochondria, critically depend on their presence. The disruption or alterations of organelle contacts are therefore associated with compromised organelle function, resulting in altered or disturbed protein and lipid synthesis, ion homeostasis and metabolism. This ultimately results in cell death. Consistent with this, changes in organelle morphology, distribution and connections are associated with multiple diseases, including diabetes, Parkinson’s disease and cardiac- and skeletal muscle dystrophy.
What is the aim of the project?
The specific aim of this project is to determine the importance of a recently identified membrane tethering protein for the regulation of insulin production and secretion from pancreatic β-cells.
What techniques and methods are used?
Standard molecular biology techniques (DNA preparation, restriction enzyme digestion, poymerase chain reaction (PCR), electrophoresis etc.) will be used to generate various fluorescently tagged versions of the membrane tethering protein. - Cell culture and transfection of clonal β-cells with the above-described fluorescent protein-vectors and siRNA. - Advanced live-cell fluorescence microscopy (confocal and total internal reflection fluorescence microscopy) to investigate the dynamic behaviour of the membrane tether under physiological and patho-physiological conditions. - Image processing, analysis and quantifications. - Insulin secretion will be measured using the Enzyme-Linked ImmunoSorbent Assay (ELISA) technique.
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student shall produce various fluorescent protein-tagged fusion proteins by using techniques mentioned above ((DNA preparation, restriction enzyme digestion, poymerase chain reaction (PCR), electrophoresis). - The maintaining of cells in culture and transfecting the cells to express the above-mentioned proteins or to deliver siRNA. - The student should be able to visualize cells by using different fluorescence microscopy techniques (confocal and total internal reflection fluorescence microscopy). - Analysing image data with the help of the knowledge the student will gain during the research, to later report results on the specifik research question.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will participate in weekly reserach seminars.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic knowledge of laboratory work (e.g. how to pipette small volumes, operate a centrifuge etc.). It is advantageous if the student has prior experience of cell culture and molecular biology techniques (PCR, electrophoresis).
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Pre-Medical students from the American-British system - Students in biomedical fields
- Feedback regulation of insulin secretion by extended synaptotagmin-1. Xie B; Nguyen PM; Idevall-Hagren O. FASEB J. 2018 Dec 27:fj201801878R. doi: 10.1096/fj.201801878R. [Epub ahead of print] PMID: 30589572
- Triggered Ca2+ influx is required for extended synaptotagmin 1-induced ER-plasma membrane tethering. Idevall-Hagren O; Lü A; Xie B; De Camilli P. EMBO J. 2015 Sep 2;34(17):2291-305. doi:10.15252/embj.201591565. Epub 2015 Jul 22. PMID: 26202220
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