Kate Barnett – Neuroscience at George Lab | Week 3

Welcome back to week three at the George Lab in UCSD! Things calmed down at the lab this week because the lab had submitted their grant renewal and was able to take a breather. I got so much closer with everyone this week – both at work and with my host family!

Monday morning, I got to pull some brain samples from freezers in the basement of Skaggs because one of the lab members, McKenzie Pavlich, was cryosectioning quite efficiently. These freezers are kept at -80°C due to the fragility of the tissues, and it is required that you wear gloves when touching anything in the freezer; dry ice freezes your skin cells and will “burn” you. I struggled with pulling out one of the boxes in the racks; it was frozen stuck. It’s critical to move quickly when pulling samples so that the organs remain frozen and preserved. Despite that, it took Lisa and me a while to chip the box free of ice. When we finally succeeded to free the box and find most of the samples, we loaded a styrofoam box full of dry ice, placed the brains inside, and hastened upstairs to stick it in McKenzie’s freezer. After, Lisa and I walked over to MTF to visit Molly Brennan to discuss what’s in the books for the George Lab in the coming weeks. In a shortened explanation, the George Lab (right now) is split between two buildings (BSB and MTF), causing resources to be split as well. The lab’s solution is to move into one building (MTF) to avoid misplaced items and time-consuming commuting. For the last several weeks, construction workers have been remodeling rooms in MTF to accommodate all of the experiments that the lab runs in BSB, reconfiguring wires, power outlets, and much more. This week’s obstacle was to communicate with all of the electricians to make sure the rooms had enough wattage in each outlet for all of the computers, self-administration boxes, lights, etc. Wrapping up with Molly, I tagged along with Brent to travel to BSB to clean out the bedding of self-administration chambers for the next round of nightly experiments. Then, I worked with Angie to re-dose a lot of oxycodone and a few cocaine syringes. A few rats had gone up in weight or weren’t affected by the oxy, so we had to empty the syringe’s contents into its original bag and then, refill the syringe with a new, stronger concentration of oxycodone. For some rats, we had to decrease the concentration of the drug they were taking because it was making them sick, which endured the same process; emptying the syringe’s contents into its original bag and then, refilling it with a lesser concentrated substance. After we refilled all of the syringes, Angie and I flushed the tubes in the chambers with ethanol to clean them out and then hooked the assigned syringes up to the chambers. The last thing I did on Monday was commuting back to MTF with Brent to clean out more bedding of self-administration chambers for the experiments running that night.

On Tuesday, I labeled more tubes with Lisa because McKenzie was (again) punching brain slices and cryosectioning like crazy! She was going through all of the tubes really quickly so I had to label quite a few sets of tubes for her but didn’t end up finishing because I went down to BSB to help Brent, Molly, and Angie clean out the bedding of self-administration chambers for the nightly oxy experiments. I also got to see the adjacent room where the vapor rats are kept and how one would weigh a rat. Because I’m not allowed to touch any of the rats, I just input all of the measurements into Brent’s computer but Angie got to learn how to place them on the scale and predict their movements which was EXHILARATING. As one could imagine, rats going through withdrawal are irritable and restless, causing them to be frantic and hard to weigh on a small scale when they’re bouncing all over the place. After weighing these animals, Brent and I traveled to MTF to weigh the remaining rats and then went up to the Wet Lab in Skaggs to make more catheters for the new shipment of rats coming in.

Wednesday morning, I did lots of running around. Literally. I bounced back and forth between Lisa, Molly, and Brent to figure out who I was going to tag along with, hustling between Skaggs, BSB, and MTF. Eventually, I came back to Skaggs to find Brent in Lisa’s area. He was trying to pull some of the missing brains from the freezers that Lisa and I searched through Monday morning. He could only find 2 of the missing brains, so he showed me how the George Lab sent different brains that were just as adequate. On a very lengthy Excel sheet was every rat that the George Lab ever had, their characteristics, and what organ(s) they had shipped or not yet shipped out of the BioBank: their rat ID, addiction index, weight, etc. Brent explained that if a brain is missing, the George Lab tries to find another brain that is almost identical to it – one with a super close addiction index, the same sex, etc. After we scrolled through what seemed like an abyss of Excel cells, we finally found a brain that matched the missing one. By that time, the lab was hosting a lab meeting via Zoom, so we traveled upstairs to the Wet Lab to dial in. In the lab meeting, I was finally (formally) introduced to the lab, which was ironic as I have been at the lab for over 2 weeks and have met everyone there already. Besides that little nugget, I got to see what a lab meeting was like! It wasn’t as formal as TV shows and movies portray it to be; it’s really just a huge easy-going conversation about what’s taking place in the lab for the rest of the week and part of next week. It’s just a game plan, really. Some members even presented data about a cohort of rats that we’ve been working with. I sat next to Angelica during the lab meeting and was mesmerized by her mounting brain slices onto slides. Then when the lab meeting wrapped up, Lani, Brent, and I walked over to BSB to meet Angie and clean out the bedding of self-administration chambers for the nightly oxy experiments.

Thursday was a relatively slow day. In the morning, Brent, Molly, and I scanned the new cohort of rats, prepping them for IV catheterization surgeries in the upcoming weeks. Brent and I also occupied Lisa’s space in the basement of Skaggs to pull more brains for McKenzie, just like Wednesday. We had more luck than Wednesday which was a feat! After lunch, I assembled catheters (only phase 1) for the new cohort of rats. It’s a simple, but precise task – mainly because the tubing is so so small. The first step to making catheters is to bend cannulae 90°. I then measured out how long the small (14cm) and large tubing (2cm) should be. When I finished cutting the tubing with a razor blade, I gathered all the small tubing and dipped the ends (of only one side) into CitriSolv, temporarily increasing the diameter of the tube so it slides more easily onto the cannulae. I also collected all of the large tubings and soaked them CitriSolv to help them slide onto the smaller tubing. I thread the small tubing over the bent end of the cannula where the tubing was flush against the start of the threaded portion of the cannula. Then, I thread the bigger tubing on the small tubing where both tubings were flush against the threaded piece of the cannula.

When I had some downtime this week, I mainly played volleyball! My host mom is a volleyball coach so she expanded my basic knowledge of the game. We’ve gone to a few different parks to play (more like pepper) as well as the beach, where we played an actual game with some friendly college kids. I caught some yummy treats at Bobboi Natural Gelato and Philz Coffee and made many trips to the marvelous Whole Foods. I’ve recently been on a huge fruit craze, so the amount of fruits I bought this week is nuts.