Vol. 2...Hard Knock [Postdoc] Life — Larry Rodriguez, PhD

General updates on what I’ve been up to since the start of the year. Cover art inspired by Hov’s 3rd album.

1st publication as a postdoc (coauthor)

At the beginning of the year, we submitted a paper to Alcohol and Alcoholism, and I’m happy to report that our paper was recently published. A little background: alcohol use disorder (AUD) differs between sexes; men are more likely to develop AUD than women, but women progress from casual use to binge and heavy alcohol use more quickly and are more likely to relapse into repetitive episodes of heavy drinking. Animal models of AUD have shown that the central amygdala (CeA) serves as a hub for processing stress and anxiety. In fact, gamma-aminobutyric acid (GABA)ergic signaling in the CeA has been shown to be involved in (alcohol dependence-induced) increases in alcohol consumption. However, previous studies have been done exclusively using males. So, we decided to see if there were sex-differences between of naïve and alcohol dependent rats. This is an important question, but surprisingly, studies on the role of sex are still lacking. Using whole-cell patch clamp electrophysiology, we looked at GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) in the CeA and found differences between the sIPSC kinetics in females and males, as well differences between the naïve and alcohol-dependent animals. We also found differences in the baseline current kinetics during the different estrous cycle stages in females. A concentration-dependent increase in sIPSC frequency is routinely found in males, but surprisingly, we found that acute alcohol (11-88 mM) had no effect on sIPSCs in naïve female rats. However, we found that 88 mM alcohol did increase the sIPSC frequency in the dependent females. Our results provide important insight into sex differences in CeA neuronal function and dysregulation with alcohol dependence and highlight the need for sex-specific considerations in the development of effective AUD treatment.

1st grant rejection as a Postdoc

“Offshore” is the most beautiful hiphop beat I’ve ever heard.

At the beginning of the year, I applied to the Burroughs Wellcome Fund’s Postdoctoral Enrichment Program (PDEP.) Well, I got the results on May 21st, and while I was a finalist, I didn’t receive the grant. I’m not gonna lie, the feeling fucking sucks; god damn it fucking sucks. WHATEVER THO. Even Jay-z lost a brick or [ninety] two. I listened to Offshore by Slaughterhouse 50 million times and now I’m back at it. The work is important, the methods are novel, I’ve got a ton of preliminary data, and the project is exciting. Best of all, I’m getting back in [biochemistry] bag. Hopefully more will be revealed/published soon!

Mouth pipetting (safely/sustainably?)

Patch clamp electrophysiology experiments are probably the only time you’re allowed to mouth pipette, and everyone has their own style. During COVID, the idea of putting the same plastic tubing in my mouth everyday kinda grossed me out, so I decided to just cut and de-filter 1 mL micropipettes each day. That mostly worked, although on particularly frustrating days, I would bite through the pipette and taste plastic from the pipette all day. Eating plastic is not great, so I started looking for alternatives, and I found a good one: bamboo straws! They’re sturdy enough so that you can apply a lot of vacuum, but malleable enough to jam in barbs and luers. Plus, they’re easy to remove and clean!

Duck thermometer

For our experiments, we need to keep our brain slices at 37°C for at least 30 minutes. Of course, there will be a little variation in water baths, which probably won’t kill the slices but can affect the quality of the cells, and thus, introduce variation in experiments. So, to minimize this, I bought a duck thermometer. Its water activated, and beeps when the temperature drifts ±1°C. Not surprisingly, the duck thermometer suggested that there are spots in the water bath that are warmer than others. ~~To fix this, I added a “circulation” system to the bath: a tube connected to the carbogen (95% O2 + 5% CO2) system that bubbles the water bath.~~ Alright, I was told by people that using carbogen to circulate the water bath is unnecessary/wasteful/stupid so maybe use an aquarium pump. MY BAD. The duck is still dope tho!

Weight-loss

In January I discovered that my homemade sub sandwiches had >1300 calories in bread alone (!!!!) and since I was ~195 lbs (what I weighed at the start of grad school) I decided to lose weight. After about 4 months, I’m down >20 lbs, and I can definitely feel a difference! I didn’t do anything too drastic, I just counted calories religiously. In fact, I’ve had in-n-out for dinner once a week, every week (3 double doubles protein style = 520 calories each!) and still managed to lose weight. I also took up running a mile everyday, but still keep my workouts to 1000 calories every morning. The main thing slowing my down right now is weekend eating; for various reasons, I haven’t been able to cook my own food on the weekend, and for the past 6 weeks, it shows. Still, its good progress, and I’m actually ok with sticking to my current diet long-term. Homemade everything is the strategy that worked best for me:

· Substitute lettuce for bread and tortillas; burgers and tacos have been protein style all year

· Homemade popcorn! 70 grams of unpopped popcorn comes out to about 210 calories and has a final volume of about 13 cups, popped. Spray with extra virgin olive oil and lightly season, and the caloric density is pretty good! I eat ~25 cups (<600 calories) of popcorn before dinner.

· Weigh and log everything!

· Substitute “I can’t believe its not butter” for most oils