Author Archives: Margo Orlen

Not Farewell but See You Soon

The past 8 weeks here at Duke this summer have been some of the most rewarding, challenging, and inspirational in my entire life. I cannot express my gratitude enough to the BSURF program, to Jason and Ron, and to my amazing mentors and lab. To anyone looking to pursue a career in research or to simply become exposed to the field, the program is an incredible summer opportunity to discover yourself and your own passions in life.

It is clear to me now that research is not for everyone as it comes with its own unique set of challenges and obstacles. It is also clear that you have to be willing to work hard and push forward in the face of failure and adversity. If you do decide to continue down this path, as I have decided, I believe the outcome is worthwhile. Seeing the work of all my peers and seeing all of our hard work come to life in the form of our posters, I know that we have reaped all of the rewards this summer had to offer. And with that said, I look forward to continuing with the lab and my studies in science as I have enjoyed every minute of this summer. I cannot wait to be back soon doing what I love most.

A Passion for Baboons

One of the reasons I applied for this fellowship program, besides my interest in biology, was to discover what I wanted to do with my life. I wanted to know if biomedical research was for me. With the help the culmination of my summer experiences, including the very insightful faculty seminars, I am now able to confidently say I believe I am following the right path for me. As each faculty member stood before us throughout the summer, I would look at all of them and say to myself this is what I want to do. A common theme a found throughout all of the speakers was a sense of passion. I could feel how every speaker radiated a certain passion for the work they are doing. And I believe this is a goal to admire and strive for: to have passion with what you do in life.

One speaker that I could feel had such a unique and admirable sense of passion was a speaker from our first week here this summer: Dr. Susan Alberts, professor of Biology and Evolutionary Anthropology. Let me start by saying I am usually more interested in cell and molecular bio topics rather than ecology or animal behavior fieldwork, yet I found myself enraptured by Dr. Alberts’ discussion on baboon social affiliation and its effects on animal fitness. First off, I found it amazing how Dr. Alberts spent countless hours in the field in Kenya dedicated to her work. Also, who knew the baboons could have such intricate social relationships. What’s more than that, the social relationships of these baboons could possibly translate into relatable outcomes observed in humans. Perhaps I found her lecture more interesting because this type of research was so different that what I am used to. Perhaps it was because baboons are pretty amazing creatures. But I also think a large aspect of her talk that captured my attention was her apparent passion for her work. Listening to her that morning, I hoped that one day I could feel the same about the work I was doing because for me there is nothing more inspiring than having passion in what you do like Dr. Alberts.

The Three Rs: Repetition, Repetition, Repetition

Something I’ve come to learn this summer is that research is all about repetition. What makes a great paper? What makes for a strong experiment? Repetition, repetition, repetition. So by now I’ve run my millionth BRET assay under a million different conditions and, to be honest, I most likely will be running a million more. As each plate gets read, I can feel my excitement rising with the promise of an interesting result in our data. Every time I run the assay there is always surprising, sometimes disappointing, data that lead us to ask more and more questions. By the end of a week full of BRET, I know every Friday afternoon on the dot after analyzing all of the data I will be planning my millionth and one BRET assay for the following week.

For me, research is well worth the repetition. I know that all of this work will pay off and hopefully there will be something significantly interesting and personally rewarding that I uncover from my numerous trials.

As I sit here now thinking about how much more BRET I could possibly run in the next 10 days of this fellowship, I can’t help but feel a twinge of sadness for the soon separation of me and my dear old BRET assay. I suppose this means that I’ve come to genuinely love the repetition of my research, which I hope to continue in the future. So with that, I am happily downloading the request form for an independent study so I can spend another cherished semester with my friend BRET.

Monkey Mind Reading

I once watched a Ted Talk a few months ago titled “Brain-to-brain communication has arrived. How we did it” featuring Dr. Miguel Nicolelis, a man with passion who astoundingly allowed a paralyzed man to kick a soccer ball at the World Cup. It all seemed so science fiction-y, especially when he showed monkeys moving robots with their thoughts. I couldn’t believe any of this was actually possible, especially right here in the labs of Duke. Although I myself am not particularly savvy in brain-machine interfaces or basically anything engineer or technology oriented, I found Ricardo’s talk especially interesting because it made me realize the ideas presented in the Ted Talk are very much real…that brain-machine interfacing is very possible and that one of our own from BSURF is working on this incredible technology.

Part of the fun of research for me is hearing about new and exciting topics other people are currently working on in the lab, and the chalk talks were a great way to facilitate the transfer of knowledge and ideas between the fellows of the BSURF program in just a few short minutes. Although I only addressed Ricardo’s talk, basically because I am baffled by monkey mind reading, I was so impressed by everyone’s work and I look forward to see the progress my peers continue to make throughout the duration of the summer.

A Week in the Life

For my on-going project, as discussed in my previous blog entry, I use the BRET assay to measure protein-protein intracellular interactions specifically by investigating the role of arrestin plays in internalization and signaling of the D2 dopamine receptor depending on the location of the receptor. Now, this assay can be modified in several manners, for example, to manipulate concentration of certain g-protein coupled receptor kinases (GRKs) or to observe the effects of different ligands or drugs in activating the receptor. Essentially, this assay can be and will be modified in a million and one ways using a million and one differing conditions, excuse my hyperbolic tone, to further uncover the mechanisms of dopamine receptor signaling and trafficking. With that being said, you best believe I have a million and one different BRET experiments that I have to conduct. So here is how the deal basically goes:

  • Monday means transfections. My mentor graciously comes on the weekend to have cells already split and ready for me to transfect on Monday with the certain receptor and protein tags. Thanks Tom, you rock for that.
  • Tuesday means splitting. After I have transfected the cells, they must be split onto a special 96-well plate in specific media. Multiple transfection plates usually means I have a long afternoon in the cell culture room.
  • Wednesday means treatment. I treat the cells on the 96-well plate with different drugs for an hour and then I can read the plate using the lovely BRET machine.
  • Thursday means repeat. Usually, I have several BRET assays staggered so I will transfect, split, and read all through the week.
  • Friday means analysis. This is the day I go through all the data I have collect and begin to interpret the results. After, I can plan out where I will go from there and what conditions I will use for the next BRET assay.

In addition to my BRET project, I am usually kept busy helping my mentor with his own experiments. Cloning, cloning, and more cloning. But I am not complaining. I love working in the Caron lab. I love that I always have something to do and I am never bored. I love the repetition as well as the novelty. I love the science behind everything and the fact that I am learning more and more everyday. Most of all…I love my lovely new friend named BRET.

BRET and I

Forty to fifty percent of therapeutic drugs today target a class of plasma membrane proteins called G-protein coupled receptors (GPCRs) (Beaulieu & Gainetdinov, 2011). The dopamine receptor is a type of GPCR that, when activated by dopamine, plays a role in reward, motivation, movement, and more. Imbalances in dopamine can result in neurological disorders such as ADHD, Parkinson’s, and schizophrenia. An overarching goal of the Caron lab is to understand the mechanisms of the dopamine receptor in order to formulate therapeutics targeting this receptor for treating these neurological disorders. Specifically, many investigators are working to improve antipsychotics for the treatment of schizophrenia by reducing side effects and increasing efficacy by targeting certain pathways of the dopamine receptor.

In order to begin to formulate these antipsychotics, it is necessary to first understand specifically the mechanisms of the dopamine D2 receptor, the target of most antipsychotics. The goal of my project essentially is to uncover the complexities behind the dopamine receptor by investigating the internalization of the D2 receptor under differing conditions, including the location of the receptor and the type of ligand used for activation, by investigating

The methodology my project employs is the BRET assay that investigates protein-protein interactions by measuring bioluminescence resonance energy transfer (BRET) between one protein tagged with a light producing enzyme, a luciferase, and another protein tagged with a fluorescing substrate acceptor, in this case a YFP. Specifically, my project investigates the interaction between the D2 receptor and beta-arrestin 2, a protein involved in receptor internalization. We are currently working on manipulating the type of GRKs and drugs used and measuring the effect this has on internalization. By the end of the summer, we hope to be one step closer to understanding the complexities of internalization and the elusive dopamine receptor.

References:
Beaulieu, J. M., & Gainetdinov, R. R. (2011). The physiology, signaling, and pharmacology of dopamine receptors. Pharmacological reviews63(1), 182-217.

An Interview with Dr. Marc Caron

One of the greatest perks of working in a biomedical research lab at Duke University is the exposure you have to some of the most accomplished and brightest minds in science. So you can imagine how I felt, a mere undergraduate student with only 2 semesters of college and 2 weeks of research under my belt, as I walked into the office of Dr. Marc Caron this week. Needless to say I was nervous, excited, awestruck, and intimidated all at the same time. Now I can say with absolute certainty that Dr. Caron is one of the smartest and most passionate scientists that I have come to know.

I asked Dr. Caron the basic questions: how he got into science, how he became interested in his field of research, and if he had any advice for an aspiring scientist based off his experiences. I came to learn of Dr. Caron’s impressive background. His interest in science began in the early 60s when he was attending the Laval University in his native country of Canada. It was during the time that he deemed the birth of biochemistry following the buzz of the development of the contraceptive pill and, subsequently, steroid metabolism. It was then that he decided to continue his studies in steroid metabolism and what he called “primitive biochemistry” at the University of Miami where he would receive his PhD. Following his time at the UM, Dr. Caron sought out a postdoctoral fellowship and found himself in the lab of Dr. Bob Lefkowitz at, ironically, Duke University where his research really began to evolve.

It was with Dr. Lefkowitz that Dr. Caron began to explore the realm of G protein-coupled receptors (GPCRs), which now comprise the most target form of proteins for therapeutic agents. Dr. Caron proceeded to spend his time attempting to purify one of these receptors with immense success in the 80s. It was 1986 when Dr. Caron and colleagues first purified and clone the beta 2-adrenergic receptor. It was then when he began to discover the incredible homology between receptors, which brings us to now…

Dr. Caron was always interested in the dopamine receptor, a GPCR. His subsequent and current research focuses on this receptor, specifically to attempt to elucidate the complexities behind this receptor and its signaling pathways. By manipulating these receptors in mouse models, Dr. Caron has developed models of ADHD, schizophrenia, and Parkinson’s. Without getting into the hairy details, the ultimate goal behind his research is to develop new or enhance existing drugs in order to develop more effective treatments for these of neurological disorders. Dr. Caron explained that 40-50% of drugs that target GPCRs are targeted to only 50 receptors, which leaves about 150 more to be tested! His excitement was evident throughout the entire span of the interview.

His overflowing enthusiasm for his research, I believe, drives the entire lab’s hard work and dedication. I look to him not only as a mentor or as my PI, but also as a fellow scientific mind with infinite aspirations and exploding passion. I was blown away by his accomplishments, his intelligence, and his zeal throughout his entire career leading up to today. As I walked out of his office, I knew I was in the right place and I was taking the right path in life to become a biomedical researcher. I can only hope that through my own perseverance I can one day become a scientist as extraordinary as Dr. Marc Caron.

BSURF 101: What is a scientist?

Taped to the wall in my lab there is a poster with several pictures, each depicting what different people think scientists do on a day to day basis (shown below).

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While cooking meth like Walter White from Breaking Bad is most likely not what scientists here at Duke probably do, I think the pictures accurately reflect some of the stereotypes of researchers from different perspectives. And to be completely honest, coming into this program, I myself was not really sure which picture in the poster really reflected what it means to be a scientist in biomedical research. Our perceptions are often influenced by depictions of scientists by the media, pop culture, or even our high school biology classes. We’ve seen the scientific breakthroughs on the headlines of the evening news. We’ve watched movies or TV shows showing the exploding flasks, three-eyed radioactive mice, and underground meth labs. We’ve done the classic frog dissections in biology lab. Defining scientific research is not so simple at after all. So with that, I hope by the end of this experience I will be able to answer: What is it that scientists really do? What does it mean to be a scientist?

This question is really important, as a huge motivating factor for doing this research experience is to solidify my passion for biomedical research and my dream of pursuing a career in academia. After one short week in the Caron lab, I can already tell you that scientists work very hard, have immense passion for their research, and also make many mistakes. In one short week I have learned and done more than I ever expected I would in such a short time. Conversely, I’ve also never been so confused, in over my head and nervous than I’ve felt in awhile. But I’m starting to think that this is what science demands. It demands knowledge and determination. Patience and practice. This summer, I expect to be challenged mentally with the new research being thrown at me. I expect to have great successes in my experiments and also great failures. I expect to learn something new everyday.

It’s not just the lab techniques and experience that I hope to have acquired by the end of this summer. It’s the experience of beginning to discover what it means to be a biomedical researcher. So when one day when my Ph.D. diploma is hanging over my lab bench, I can then fully answer what it really means to be a scientist.