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Investigation of neuroprotective effects of mitochondria-targeted drugs in experimental stroke models
Lithuanian University of Health Sciences
Laboratory of Biochemistry, Neuroscience Institute, Medical Academy
Prof. V. Borutaitė
Prof. V. Borutaitė, Dr. P. Čižas, D. Umbrasas
Type of Research Project
- Basic science
What is the background of the project?
Ischemic stroke is one of the main causes of death and disability worldwide. The severity of neurological defects is thought to be dependent on neuronal death in ischemia-affected areas of the brain. Mitochondria are considered as key payers in ischemic brain damage though precise molecular mechanisms of ischemic brain damage are not entirely clear. One of the mechanisms of ischemia-induced neuronal damage involves opening of mitochondrial permeability transition pore which leads to depletion of cellular ATP, loss of pro-apoptotic molecules from mitochondria and induction of necrosis or apoptosis in the ischemia-affected brain. The regulation of these pores in mitochondria is not well understood but may involve modulation of activities of cyclophilin D and Complex I of the mitochondrial electron transfer system. The aim of the current project is to investigate whether various chemical compounds acting on mitochondrial Complex I or other components of the mitochondrial electron transfer and phosphorylation system may exert neuroprotective effects during brain ischemia.
What is the aim of the project?
To investigate the effects of potential neuroprotective compounds on brain mitochondrial functions and neuronal viability.
What techniques and methods are used?
Preparation of neuronal cell cultures and their maintenance. Methods of detection of cell death and viability. After homogenization of brain tissue and centrifugation of isolated brain mitochondria, total mitochondrial protein is determined by the modified Biuret method. Measurement of mitochondrial functions by using high-resolution oxygraph. Assessment of mitochondrial calcium handling capacity fluorimetrically (using Fluorescence Spectrometer and fluorescent dyes): spontaneous release of Ca2+ causes an increase in fluorescence indicating mitochondrial permeability transition pore opening referred as calcium retention capacity.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
What are the tasks expected to be accomplished by the student?
Students will join and perform specific laboratory work and research tasks which will include performing experimental procedures under supervision such as: • extracting and homogenizing brain tissue; • isolating mitochondria; • determining total mitochondrial protein; • determining calcium retention capacity in isolated mitochondria; • collection and analysis of experimental data with SPSS.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Students will be provided with relevant scientific papers and will discuss the tasks, experimental work and results obtained with supervisors.
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 - The student will prepare a scientific report - The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
To be familiar with general rules and techniques used in the biochemical laboratory (pipetting, preparation of solutions, calculation of concentrations of substances in solutions, pH measurement, work with a spectrophotometer, etc.) • Subjects passed: chemistry, biochemistry; • Previous experience with: work in biochemical laboratory. We also expect that the student will attend at least 80% of the research exchange, be responsible for his/her actions at the practice and be accountable to the supervisor and LORE/NORE.
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
- Borutaite V; Toleikis A; Brown GC. In the eye of the storm: mitochondrial damage during heart and brain ischaemia. FEBS J. 2013; 280: 4999-5014.
- Rekuviene E; Ivanoviene L; Borutaite V; Morkuniene R. Rotenone decreases ischemia-induced injury by inhibiting mitochondrial permeability transition in mature brains. Neurosci Lett. 2017; 653:45-50.
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