“Sun’s gettin’ shinery, to spotlight the finery, Spring, Spring, Spring.” – from my all-time favorite musical, Seven Brides for Seven Brothers. How I love Spring Quarter (and corny old-fashioned musicals)

The rest of this blog will be research focused, I am really excited on finally some conclusive understanding of the results from several experiments! 🙂

Abstract: Many neurodegenerative diseases, including Alzheimer’s Disease, Frontotemporal Dementia, and Niemann Pick Type C, are characterized by the histological finding of  intracellular tau aggregates termed neurofibrillary tangles (NFTs) in the CNS. We hypothesize that the amount of NFTs in hippocampal tissue is the result of dynamics between the clearance, production, and accumulation of aggregation prone proteins, particularly phosphorylated Tau. To test that idea and quantify the extent of tauopathy of aged 3xTg mice, we used phosphoTau (PHF1) immunofluorescence to observe NFTs and Hoechst to fluorescently label nuclei. Mice were either treated or left untreated with compounds that modulate the biochemical pathways of clearance or production of NFTs. The chaperone proteins Hsp70 and Hsp90 regulate tau degradation and clearance within the neuron. By treating mice with Hsp70 inhibitor methylene blue, or with Hsp90 inhibitor celastrol, we found that perturbations of the chaperones’ activity significantly increased the percentage of NFT+ cells found in the hippocampal areas of the mice brain. Phosphorylated Tau production is regulated by the activity of several kinases, of which Cdk5 and GSK3-β are of particular pathological interest. The kinase inhibitor ___ specifically reduces the activity of Cdk5 and GSK3-β. We therefore propose that treatment of mice with ___ will significantly reduce tauopathy within the hippocampus by inhibiting phosphorylation of tau and impacting the production of NFTs. In this work, immunofluorescent microscopy imaging of tauopathies for each treatment was achieved and new techniques of quantifying the extent of tauopathy developed.

Last year, I finished all of the methylene blue experiments. The past two quarters, I have been working on the celastrol portion of the research. Based upon the results of these two major projects I have formulated the abstract above and am now in the process of immunofluorescent microscopy of the ___ treated mice. ___ is a brand new compound synthesized by another member of the Kosik lab to mimic the activity of a specific micro-RNA that targeted the same binding site on CDK5 and BSK3-β.

Over the course of four quarters, the shape of this research project has developed, molded, and flipped around according to the results acquired. Hypotheses have changed and new questions have been developed. Reflecting upon my own growth throughout the past year and also the development of this research project that has broadened its scope beyond which I could have imagined, I am incredibly grateful for the opportunity I have been given to engage in research through the EUREKA internship. I am beginning to realize that the only way to really gain understanding and an appreciation for scientific research is to fully immerse oneself in its whirlpool of utter confusion, in the hope of experiencing its priceless ah-ha moments.