First Week in the Smith Lab

Hello everyone! I'm Jaeyoung Kang, and I'll be updating all of you regularly on what's happening during my time in the Smith Lab at Arizona State University.

The Smith Lab works on utilizing olfactory cues to condition insects with Pavlovian conditioning for the purpose of better understanding the cellular and molecular changes in the brain during moments of learning and memory. Specifically, I'll be working on conditioning honey bees (Apis mellifera) using the PER (Proboscis Extension Reflex), and I will be working with Dr. Irina Sinakevitch to image the bees to assess for the presence of GABA receptors. The work I'm doing is a part of Dr. Irina Sinakevitch's big project to make a link between the GABA receptors and conditioning in the bees - should be pretty cool.

To tell you about what I'm doing, I should quickly break down the procedure. So the first step is to collect the honey bees from the hive. It's already been the first week, and I've already had a chance to catch bees straight from their hives! The bees are not dangerous - provided you don't swat them or otherwise do something dumb, it's difficult to get stung. I caught them right from the hive without any protective gear. I have a video of my mentor, Dr. Brian Smith, demonstrating the technique - I'll post a video soon of me doing it! Don't try this at home though - wild bees in Arizona are most likely aggressive, Africanized killer bees, and will be much easier to provoke.

After catching the bees, you have to condition the bees. Basically, that involves sedating them with ice, harnessing them in place, and then exposing them to various scents while feeding them sugar water using their Proboscis Extension Reflex. I got to make harnesses for the bees, which was awesome (picture in next post)! The bees have receptors on their antennae, and when you put sugar water near the antennae, the bees will automatically bring out their proboscis, and consume the sugar water given. So the conditioning involves exposing them to scents while feeding them, and after the conditioning, seeing if they exhibit the reflex when exposed to the scent, without the sugar water. There are variations that can be done with this, but more on that another time.

Once you do this step, then you get to the imaging, which uses immunohistochemistry and confocal microscopy. I haven't started this part yet - I should be starting soon. I will be taking the conditioned bees, and injecting dyes with fluorescent antibodies into their brains. Then, I will use confocal microscopy, which shines lasers into the brain, to "light up" the regions the antibodies attached to. The antibodies bind specifically to GABA receptors, so when I get pictures of images from the confocal microscopy procedure, the colorful areas should be areas with GABA receptors. I will be assessing differences in the presence of these GABA receptors between the conditioned and unconditioned bees. For those unfamiliar with GABA, it's the main inhibitory neurotransmitter of the brain - assessing receptors for GABA should be a compelling avenue of research.

Currently, to inject the dyes, the lab uses pressure injection technology, which injects the dye into the entire brain. However, I eventually will need to do localized injections for just a few neurons, so I'll be working on getting the current injection technology to work. I'll have to tinker around with the machine's calibrations to get it "just right." For the rest of this month, I should be working on the injection techniques and technology, and getting familiarity with the conditioning process. To help me practice, Dr. Smith is having me sit in on an undergraduate class currently doing Pavlovian conditioning labs with the honey bees and the PER. On Thursday, however, he had me work with his Ph.D student, Chris Jernigan, to help set up the Pavlovian conditioning lab for the class. I'm also working with Dr. Irina Sinakevitch to practice pressure injection techniques and dissections of the bee brains. I've got to be ready to work in her project, so I'n practicing the techniques needed to do injections and dissections, and I'm also studying the anatomy of the bee brain closely.

Super fun stuff!