Axons Weeks 4-6: Too many projects, too little time

The last few weeks have gone by in a blur. We’ve done a lot of experiments and analyzed a ton of data. The students in our lab have bonded as well; we’ve taken some weekend trips to Richmond and started running together in Colonial Williamsburg on Mondays. It’s been really fun getting to know them this summer! I also went on vacation for a week and a half (which was awesome), so I haven’t been able to post anything for a while. The following are some highlights from weeks 4-6 of my summer.

In my last post, I talked about a taxol experiment that tested for cell death. I finished analyzing the data, but the results were inconclusive. This might be the most frustrating outcome I’ve experienced in research. Each explant plate experiment takes about 2 weeks to complete, so it’s disappointing when you don’t get significant results from all that work. There is plenty more for me to do though, and as they say, there’s no sense in crying over spilled milk. Or neurons, I guess.

I did a DMSO dose-response experiment as well. Dimethyl sulfoxide (DMSO) is a colorless liquid that is used as a solvent for many of the drugs we use on neurons in our lab. Too much DMSO can cause axon degeneration, so we need to make sure that the amount in the drugs isn’t affecting our results. My results for that experiment looked really good. The DMSO concentration that the drugs contain looked very similar to the controls, which is exactly what we want to see.

I’ve started piecing together a pipeline for making SARM knockout zebra finches based on papers from other labs that have made transgenic finches. We probably won’t get to most of it until this coming year since we’re still working on getting the virus ready, but I’ve made a little progress this summer. In order to make SARM knockout finches, you first need the virus that gets rid of the SARM gene. That’s what Karen has been working on, and she’s doing a great job! Next, you need to infect the embryo with the virus at a very early age (like “I’m just a clump of cells” age). In order to do that, you need to open a tiny hole in the eggshell, inject the virus, and then use another tiny piece of eggshell to cover the hole. This is the part I’m trying to perfect so I can infect when the virus is ready. Here’s a picture of an egg with the hole cut out, as seen through the dissecting microscope:

That little white spot is the embryo!

The little white spot is the embryo!

I’ve had mixed success with these eggs. The hole doesn’t seem to affect their development, but I’ve had trouble getting the shell covers to stay in place. If they fall off in the incubator, the embryo dies. I’ve also pierced some embryos with micropipettes to see if they’ll develop normally. Most of those have died so far, but I’m still new to using the micropipettes. I’m hoping that practice will make perfect.