Flores Gonzalez : Observing Morphological Changes in Microglial Cells Treated with LPS and a Potent TAK1 Inhibitor
Observing Morphological Changes in Microglia Cells Treated with LPS and a Potent TAK1
Daniela Flores Gonzalez
Indiana University-Purdue University Indianapolis (IUPUI), IU School of Medicine
Daniela Flores Gonzalez, North Central High School, Indianapolis, IN; Leander Quiroz, Carleton
College, Northfield, MN; Casandra Carrillo, Teri Belecky-Adams, Carleton College, Northfield,
MN, Department of Biology, Indiana University-Purdue University
Diabetic retinopathy (DR) is a complication of diabetes mellitus and the leading cause of blindness in working-age adults. Microglia are immune cells present in the CNS that when activated for long periods of time can cause damage to the cell and blindness in patients. TAK1 is a downstream signaling pathway that if inhibited, could become a treatment for DR.
Microglia cultures were grown on 6-well plates and then treated with 100pM of TAKinib or Vehicle for 24 hours before being treated with 10ng LPS or Vehicle for 1 hour. Cells were fixed with 4% paraformaldehyde. Cells were stained with cell mask orange cytoplasm stain and a nuclear label, Hoechst and imaged on a fluorescent microscope at 40X magnification. Cell morphology in all treatment groups were analyzed to conclude perimeter using ImageJ software.
Untreated microglia, when compared across all treatment groups, produced a higher degree of significance compared to microglia treatments compared across all groups. LPS-treated microglia combined with another treatment resulted in a higher perimeter compared to untreated, Vehicle, and LPS-treated alone.
Since a higher perimeter was observed in DMSO+10ng LPS treatment compared to 100pM TAKinib+10ng LPS treatment, this could indicate that there is a more prominent activation effect on microglia given LPS treatment than when given a TAK1 specific inhibitor. A more thorough quantification of cell morphology is necessary to conclude that cell activation occurs in cells.
Translational/Human Health Impact:
The inhibition of TAK1 as a possible treatment for DR is becoming increasingly studied for its ability to control microglia activation. There is also a growing need for more accurate and effective methods for detecting cell activation not only for research but also for diagnosing DR in its earlier stages and preventing a patient’s further vision loss.