Glial cells surround neurons within the brain and serve many functions. The astrocyte can metabolize neurotransmitters, regulate blood flow, and participate in the formation of new synapses. Microglia are important in the brain’s response to disease. Astrocytes and microglia will respond to the immune attack by synthesizing inducible nitric oxide synthase (iNOS), an enzyme that produces nitric oxide (NO•), a highly reactive molecule.
The project will entail studying the modulation of the synthesis of iNOS in vitro, using cells prepared from 2d rat cerebra. Lipopolysaccharide (LPS) a component of bacterial cell walls will be used to activate the cells to synthesize iNOS. Previously, we have found that the anti-depressant fluoxetine (Prozac) and the tri-cyclic anti-depressant nortriptyline will increase the release of NO• and synthesis of iNOS induced by LPS. This effect has been observed in mixed glial cultures, where astrocytes and microglia are together. The next step is to produce purified astrocytes and microglia to determine whether the effect of fluoxetine and nortriptyline is on the astrocyte, microglia, or both.
We have begun to explore the possibility that the anti-depressants affect the release of cytokines from the cells. Cytokines such as transforming growth factor beta 1 (TGF-ß1) can inhibit the induction of iNOS synthesis in microglia treated with LPS.
The role of the student: The student will learn to prepare the mixed glial cell cultures, nitrite assay, immunocytochemistry to identify % purity of our purified cultures, protein assay, and western blot analysis.
The student should have completed at a minimum: Introduction to Brain and Behavior and Bio 182. Preference will be given to those students who have also taken Neurochemistry because much of what is taught in the course is important in understanding the research during summer.
Number of Student Researchers
Applications open on 01/15/2017 and close on 02/07/2017