Kidney Injury Workshop-Day 2

I am currently typing this in bed.  I left Vanderbilt and immediately came back to my AirBnb.  I AM EXHAUSTED.  I learned so much today that I feel like my brain is going to explode.

7:30-8:00 a.m.:  Breakfast time.  Just as Michael Phelps does before a swim, I carb loaded.  However, I stuffed my face with scones and croissants.

8:00- 9:00 a.m.:  Dr. Agnes Fogo came and gave an overview of mouse renal pathology.  Even though I work with the kidney, my knowledge of pathology is extremely lacking, so this overview was really useful.  She spent more time focusing on the different cell types in the kidney and what their capabilities are, and how they react to injury/presence of antigens.  It was interesting to think about what cells were more likely to be affected by certain types of injury, and the kind of damage I should be looking for in tissues from my model.  

9:00-9:30 a.m.: One of the things I've realized in my time working with mice, is that data are highly variable, and results are not always reproducible from mouse strain to mouse strain.  We had a veterinarian pathologist come talk to us about some of these issues.  She focused mainly on how sex, strain, and circadian/seasonal changes can affect experimental outcomes.  One of the most interesting findings she brought up was a study published in Nature.  In this study they found that mice were more likely to grimace (a sign of distress) when being handled by females than males.  The researchers also tried putting females in shirts worn by males, and having them carry swabs from under the arms of males while handling mice.  It was concluded that the male scent was enough to not make the mice grimace.  I also learned that it is very important to order all your mice from one vendor, and that you should NEVER order control mice if you're breeding genetically modified mice in house.  

9:30-10:30 a.m.:  This session focused on ways of measuring kidney function and injury in experimental models.  For this, we usually use blood urea nitrogen (BUN) and serum creatinine (SCr) assays.  These are waste products that build up in the blood when the kidney's filtering function is not working properly.  In our lab, we've been having problems with these assays.  It was a relief to find out that we are not the only ones.  Both BUN and SCr have their pros and cons, and usually do not corroborate with each other well.  I think for my project, that it may be best to measure kidney function by monitoring glomerular filtration rate (filtering of blood through the kidney).  This process utilizes a small, non-protein bound molecule that is freely filtered through the kidney.  GFR can be measured in both the blood and urine.  Since I am proficient at collecting urine from mice, I think this is going to be our way to go.

10:30-12:30 p.m.: One of my favorite techniques to do in the lab is histochemistry, or staining slices of kidney tissue to get an idea of injury that is occurring as well as presence of proteins of interest.   These two hours were spent going over different techniques used to look at kidney injury, and I learned a lot of cool tips and tricks to improve the stainings we currently do in our lab.

12:30-4:00 p.m.: Hands-on time again!  At this point, i was already exhausted.  However, I love any chance I get to practice new techniques. During this time, we learned how to whole body perfuse a mouse.  This technique was really delicate and intricate, which means I messed it up the first time.  The purpose of a whole body perfusion is to flush out blood from the body, and then replace the blood with a fixative to preserve all the organs properly.  To ensure that we were perfusing correctly, we got to use a blue dye.  So, if done correctly, THE WHOLE MOUSE WOULD TURN BLUE.  And that's exactly what happened on my second attempt.  First, the heart, then the liver, then the kidney.  We even opened up the skull to reveal a BLUE BRAIN.  It was so cool.