I see I am getting behind on updating this blog!
Sleep, eat, eat, sleep, cuddle, watch soccer, sleep, eat, laugh, bake cookies, shop, sleep, read novels, sleep, repeat — that about sums up my winter break in a simple string of words.
Study, sleep, study, go to class, research, study, eat, study, research, applications, RA meetings, study, sleep, repeat — note the scarcity of the word sleep — these represent my first two weeks of winter quarter.
Now you, my reader, are caught up on my life between blogs! It has been pretty busy, but after this friday when scholarships and applications for next year are submitted, my load will lighten up 🙂
My research project is moving forward, and the data from last quarter is so motivating. Although the results of the Celastrol treatments suggest the opposite effect upon tauopathies of what Israel and I were expecting from our hypothesis, there is a definite trend. In research I have learned that this characteristic of results – consistency and a distinct trend as one variable is changed – is the most valuable result. A trend can be studied – it can be questioned and tested. Random inconsistent results are the only ones to be feared.
I am now in the process of digitally quantifying the images from the Celastrol experiments from fall quarter. However, based upon a rough estimate of the fluorescent signal of tauopathies by eye, it appears that treatment with Celastrol had an identical increasing effect upon tauopathies as Methylene Blue. (Remember that the presence of Hsp90 in the cell has been shown to regulate the levels of Hsp70 through a negative feedback mechanism). Therefore, inhibition of Hsp70 and Hsp90 in separate models had an identical effect – an increase in tauopathies. These results suggest the hypothesis that if the cell undergoes stress relating to any alteration of Hsp70/Hsp90 activity, then tauopathies will increase within that cell, regardless of whether Hsp70 or Hsp90 activity is the protein targeted by the drug.
Now, these conclusions are dependent upon the assumption that Methylene blue inhibits Hsp70 (spiking Hsp90 levels) in neurons, and Celastrol inhibits Hsp90 (spiking Hsp70 levels) in neurons. To ensure that this is true, I have just finished designing an antibody staining of Hsp70 in these same Celastrol tissues. By choosing samples with high, low, and no signs of tauopathies from treated and untreated mice, and then staining them with antibodies targeting Hsp70, I can answer two questions: 1) Do tissues with high levels of tauopathies also display high levels of Hsp70 indicating a correlation between the two? and 2) Are Hsp70 levels dependent upon (augmented by) high doses of celastrol? In other words, is the celastrol increasing Hsp70 levels?
Simultaneously, I have chosen samples identical to the ones chosen for the Hsp70 staining to determine the levels of the protein LC3B, a protein associated with the level of autophagy (degradation of tau in these neurons by lysosomes). Last year, we performed a similar staining with the Methylene Blue tissue samples, and found that an increased dosage of MB actually increased the levels of LC3B, regardless of the fact that tauopathies increased. It will be interesting to see whether Celastrol has the same effect. If the results illustrate an increase in autophagy, then the consistency of both the MB and Celastrol experiments will provide a strong case for the hypothesis that increasing lysosomal activity will actually increase the level of tauopathies within neurons in the hippocampus of the brain.
Finally, because this post is steadily lengthening, we have proposed that Methylene Blue and Celastrol may have the ability to induce tauopathies. This new hypothesis will be tested by designing an experiment to determine the effect of Methylene Blue and Celastrol upon tauopathy levels in wild type mice.
Dr. Kosik has given my mentor and I permission to begin writing a research paper to be published about this project. For this reason, I am looking forward to the exciting possibility of staying here at UCSB this summer and having the time to catalyze that process. But there is always a catch, I need funding to pay for living expenses here. Hence the applications mentioned above. Back to writing my proposal for the Crowe Family Summer Research Scholarship. Wish me luck!