Struggles: Forget Everything You Know (Weeks 2 and 3 in the Lab)

So I found out I need to forget a lot of things I thought I knew.

Week 2 was mostly normal - I practiced behavioral conditioning on the bees with Chris. I thought I knew how to harness the bees, but I found out I needed more practice the hard way. I got stung on the finger. It was mostly okay - the finger just ballooned up, and I had an extra struggle using it for anything. Otherwise, fine.

Week 3 was really important. I finally started the neuroanatomical experiment with Dr. Irina Sinakevitch. This is where I learned for sure to "forget everything I know." Basically, a lot of the stuff I talked about 3 posts ago is not correct - I thought I knew how the procedure worked, and I didn't. Luckily I got the breakdown again, and now, I know for sure what I'm doing. The lab already figured out that GABAa receptors, which process inhibitory information, are in the antennal lobes in the bee brain, and are important for behavioral conditioning. My job is to figure out which neurons in the antennal lobes have the receptors. I was wrong in the sense that the neuroanatomical experiment has nothing to do with comparing conditioned and unconditoned bees. Well, at least I know what I'm doing for sure now.

What else did I relearn? Injecting the antibodies is only one part of the procedure, and the antibodies aren't inside the dye. The purpose of the dye is to make sure that when we do inject the antibodies, we can see the antibodies that attached to the GABAa receptors in the antennal lobes. Basically, without the dye, we would see the antibodies everywhere there were GABAa receptors, when we only really care about what's going on in the antennal lobes. The pressure injection techniques the lab uses are for injecting the dye into the right parts of the brain, and the current injection techniques that I'll be getting to work sooner or later serve the same purpose, except it is even more localized, down to specific neurons.

I also finally learned why I was doing behavioral conditioning: The behavioral experiment involves me conditioning bees with low versus normal levels of GABAa receptors, to see differences. Again, I had to forget what I thought I knew; the conditioning was not for the purpose of comparing conditioned and unconditioned bees.

So I think I've covered enough of my blunders. I'll defend myself by saying it was only the first few days, and now, I have a much better idea of what's going on.

Now for the cool parts: I dissected a bee's head! I was helping Dr. Sinakevitch with dye injections, and to do this, we have to open the bee's head to inject into the brain. I actually opened a bee's head, and I injected into the antennal lobes and the projection neurons! Amazingly, the bee is alive and well during the whole procedure of opening, injecting, and closing the head. I'll try to get pics if I do this again!

I also found out our lab is submitting a research poster to an imaging conference. I'll be second author, which is pretty cool. Wondering if I can use this poster for SRP presentations?

And finally, my abstract. This abstract is the best description of what's going on. For a quick summary, use this abstract - I'm sure it's completely correct. Don't use my post from eons ago.

The abstract:

My work in the Smith Lab at Arizona State University is to be part of an ongoing project: “The Apis mellifera GABAa Receptor (Rdl) in the Olfactory Circuit in the Honey Bee Brain.”

The Smith Lab studies the fundamental question to investigate processes of learning and memory associated with olfactory cues.  The lab uses behavioral analyses, fluorescence microscopy and molecular biology techniques, using insect and mouse model animals. Through previous investigations, the Smith Lab established the importance of inhibitory signals, particularly from GABAa receptors, in the honey bee’s ability to process olfactory information. The Smith Lab developed the knockdown experiments in which the GABAa receptor level can be decreased. In the behavioral paradigm, the Smith Lab notably takes advantage of the Proboscis Extension feeding Reflex (PER) while conditioning bees to associate certain scents (odor) with the presence of food (sucrose). My part in the behavioral experiments consisted of observing the changes in the bees’ behavior toward odors by using the PER technique on bees with low versus normal levels of GABAa receptors.

Processing of olfactory information in bee brains begins in the antennal lobe (AL).  The presence of GABAa receptors in the AL was already established in the Smith Lab. However, it was not known what type of antennal lobe neurons express the receptor. My part in the neuroanatomical experiment consisted of identifying AL neurons that express the GABAa receptor by using both neuronal tracers to label different types of AL neurons and immunofluorescence techniques with anti-GABAa receptor antibodies.