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Discrimination of roles of activated microglia from those of blood-borne cells in ischemic brains of the rat.
Japan (IFMSA-Japan) - Ehime University, Ehime
Department of Molecular and Cellular Physiology, Graduate School of Medicine, Ehime University, Toon, Ehime, Japan.
Type of Research Project
- Basic science
What is the background of the project?
Microglia, one type of glial cells,areresident macrophage of the brain.Microglia rapidly become activated in rat stroke model brains while releasing various proinflammatory mediators and reactive oxygen species (ROS) or eliminating degenerating cells and tissues. They also release a number of neuroprotective factors such as IGF-1. Because of thus complicated functions, it has not been settled whether the activated microglia are the ameliorative cells for ischemic brains.Furthermore, there is another serious problem for the study of microglia in ischemic brains: the similarity of microglia and infiltrated blood-borne macrophages.
What is the aim of the project?
In order to evaluate whether activated microglia are aggravating or ameliorating for ischemic brains, we will use some agents that suppress or activate microglial cell functions in ischemic brains as pharmacological tools such as bromovalerylurea or cytokines IL3 and GM-CSF. The effects of the agents will be evaluated by behavioral tests and immunohistochemical staining.
What techniques and methods are used?
In this project, we will prepare a rat stroke model that is transient middle cerebral artery occlusion (tMCAO) by the use of immunohistochemical staining, flow cytometry and other techniques. Then we will use some agents that suppress or activate microglial cell functions in ischemic brains as pharmacological tools such as bromovalerylurea or cytokines IL3 and GM- CSF. The effects of the agents will be evaluated by behavioral tests and immunohistochemical staining. Mainly, the roles of microglia from those of blood-borne cells in a rat stroke model will be observed. All other techniques include Western blotting, qPCR (quantitative polymerase chain reaction), FACS (flow cytometry), behavioral tests (Morris water maze, passive avoidance, elevated plus maze, rota rod, open field), electroencephalography, nitric oxide measurement, cell cultures and assay of the cell metabolism.
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
What are the tasks expected to be accomplished by the student?
Students will be able to accomplish many experiments by themselves, including biochemical assays, such as western blotting, qPCR (quantitative polymerase chain reaction), nitric oxide measurement, and physiological assays, such as electroencephalography, HPLC (high performance liquid chromatography), and behavioral tests, such as Morris water maze, passive avoidance, open field, elevated-plus maze, rota-rod. Basic knowledge about our projects and neurochemistry, biochemistry, physiology will be acquired in this project.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Yes. Preliminary readings, lectures and/or seminars for the students will be given.
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’s name will be mentioned in a future publication
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic knowledge about physiology, biology, pharmacology are required.
Are there any legal limitations in the student’s involvement
Students are required to have a training lecture for animal experiment. This lecture will be provided in Ehime University before starting the experiment.
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Students in biomedical fields
- Iba1(+)/NG2(+) macrophage-like cells expressing a variety of neuroprotective factors ameliorate ischemic damage of the brain. Smirkin A et al. 2010 DOI:10.1038/jcbfm.2009.233
- Expression of MCP-1 and fractalkine on endothelial cells and astrocytes may contribute to the invasion and migration of brain macrophages in ischemic rat brain lesions. Tei N et al.2013 doi: 10.1002/jnr.23202. Epub 2013 Feb 12.
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