Author Archives: George Romero

The Value of Communication

While I have only worked in Dr. Mooney’s lab for a short time, I feel like I have learned an amount of knowledge far vaster than what I expected from a brief eight weeks of lab work. From learning lab techniques like injection surgeries, histology and immunostaining, and confocal microscopy to studying the neural circuits behind song learning and vocalization, this summer has been one of the most intellectually stimulating periods of my life.

Something that I came to truly realize in the last two months was how incredible nature is, specifically relating to how some biological systems are conserved throughout life. The fact that we are able to use model systems, like zebra finches, to discover and study systems within the human body is indicative of the efficiency and organization of nature. Thus, I believe it gives these organisms an innate value which deserves the respect of researchers and scientists. 

In working with the neural circuits behind learned vocalization in zebra finch, I have gained a new appreciation for the complexity behind communication. How we learn to communicate, from a neuroscience viewpoint, is perplexing and necessitates years of future research and study. But on a larger scale, my time in BSURF has emphasized the importance of good communication. As scientists, it is vital that we can communicate our findings, no matter how complex, in a way that is accessible to diverse communities and larger society. Educating others that our experimental findings are important and worth studying is what gives value to our research. For me, the intersection of my BSURF experience and working in a lab that studies learned vocalization and communication has set me down a lifelong commitment to exploring all facets of communication: the scientific and practical aspects of effective communication.

Can Technology X successfully target and manipulate dopaminergic neurons within the zebra finch?

George Romero

Mentors: Jiaxuan Qi, Richard Mooney, Ph.D.

Department of Neurobiology – Duke University School of Medicine

Previously studied neurons and cell types within the brain have been characterized using methods that are considerably slow, inefficient, and expensive. To present, there lacks a cell type targeting technology that is both widely accessible and efficient. Technology X is a novel RNA-based cell type targeting technology that can target and manipulate specific cell types within the brain. In vivo functionality of Technology X has not been completely confirmed and is under testing across multiple cell types and between different model organisms. We tested the hypothesis that Technology X will successfully target and express within dopaminergic neurons of the zebra finch. Technology X, tagged with the green fluorescent protein (GFP) indicator, was virally injected into the VTA and SNc (regions densely populated with dopaminergic neurons) of the zebra finch. In our results, we plan to see widespread and exclusive expression of GFP within the dopaminergic cells of the VTA and SNc when viewed under confocal microscopy. This would corroborate that Technology X is capable of successfully targeting and manipulating specific cell types within the brain.

The Impacts of Our Research

Now that we have passed the halfway point of BSURF, I realize more and more that science is an art of communication. While the experiments we conduct and the data we collect have their own intrinsic value, as Dr. Grunwald constantly stresses, it means nothing if we are unable to communicate it to others. After our week of chalk talks, I am truly amazed and inspired by my fellow BSURFers at their ability to communicate their research to the public – everyone really did a fantastic job at summarizing their projects!

With that said, one chalk talk that particularly stood out to me was Xitlali’s with her research into the impact of urban development on aquatic insects. What initially drew me into her project was how involved it was. In her talk she described her methods – how she goes off into Ellerbe or New Hope Creek and sets up sticky traps to collect insects. I’ve always wanted to do some sort of field research, so I must say I am a little jealous!

But the aspect of her project that truly sparked my interest was how related it was to the local community. Oftentimes in our research, we can become far removed from the fruits of our labor. Don’t get me wrong – researching the structure of specific enzymes and exploring the molecular pathway of important proteins is fascinating. However, it can be sometimes difficult to actually observe how that research is applied to larger society – to see how it contributes to concrete change in the community. 

That is something I loved about Xitlali’s talk and project. It was evident how her analysis of insect populations in the area could be used to inform policy changes in Durham and the surrounding area. If I were in her shoes, I would find satisfaction – almost pride – in knowing that my research contributed to concrete and beneficial change in my own community. Finally, her project affirms that science is interdisciplinary – that it can involve the intersection and collaboration between fields such as ecology, public policy, government, and health.

Procedures, Practice, and Papers

Alright – a day in the life at the Mooney Lab. First off, I’d like to say that there isn’t really one “typical” day or set routine in the lab. The day pretty much depends on the experiments you want to conduct that day, or if you have to continue procedures related to previous experiments you conducted earlier. That being said, I’ll do my best to give a good picture of what my day is like in the lab.

I typically arrive at 9:30 AM or 10:30 AM (depending if I have BSURF-related activities beforehand). Once there, I usually have a quick meeting with my PhD candidate mentor, Jiaxuan, about anything I need to complete that day, throughout the week, or for my summer project in general. If she doesn’t have any specific tasks for me to do at that moment, the day is really up to me. Sometimes, I go to my desk and read papers related to the research at Mooney Lab (Jiaxuan and I meet weekly for a small journal club). Other times, I simply do work not necessarily related to my project, but to the fields of neurobiology and neuroscience in general. However, most often, I’m practicing the different procedures and protocols necessary to conduct my project.

My project requires me to conduct an injection surgery on zebra finches. Essentially, it is a minimally invasive operation in which I open small sections of the skull in order to access different regions of the zebra finch’s brain. Then, using a variety of equipment, tools, and measurements, I inject a variety of substances into the brain. For the purposes of practicing, I typically inject “red beads” – a fluorescent dye that can be easily distinguished under a confocal microscope (as well as the naked eye). These surgeries typically last around 3-5 hours (depends on how many injections need to be conducted). 

Other procedures that I am currently getting the hang of are, for example, perfusions (the process of replacing the blood of a sacked organism with preserving chemicals) and immunostaining (the process and use of different antibodies and reagents to stain brain tissues). Between conducting these different protocols, reading papers, and getting a better grasp of neurobiology in general, my days are usually kept quite busy. Typically, my day ends at around 5 PM (but again, science sometimes requires working strange hours!).

The Tough Questions of Research

Complex questions never have one simple answer. Similarly, the hardest questions never offer the most satisfying answers. Reflecting on my conversation with my PI, Dr. Mooney, I realized this remains true for many questions in the field of research. We spent brief moments discussing the typical “interview” questions: how he received his PhD in Biology from CalTech or how his interest in music and science intersected in the field of neuroscience. 

The majority of our talk, however, was centered around the more complicated questions of research – questions that I believe to have multiple and possibly conflicting answers. A topic that arose in our conversation was related to the pursuit of a research project. Often, science can be unforgiving: experiments fail or data does not reflect a desired conclusion. How do you determine when to move on from a project? At what point do you determine that it is time for a new idea? Dr. Mooney, who acknowledges his tenacity and ability to commit himself to ideas and projects, explained that these are existential questions that researchers constantly grapple with. There isn’t a set “threshold” or set of criteria that scientists can refer to when considering the viability of their projects. He explained that it requires considering the unique factors of that situation – weighing the pros and cons of continuing, or not continuing, a research idea. 

We also discussed the ethics of research. Specifically, how do we as scientists grapple with the ethics and implications of animal research? Dr. Mooney explained that this is one of the most fundamental and challenging questions of research – a question that he finds himself pondering more and more as he moves further in his career. Animals, as he believes, are valuable for their own sake. Each has a unique way of sensing and perceiving the world around them – a fact that makes them valuable in themselves. Yet, he also acknowledges their value in terms of research and better understanding the human condition. Specifically in the field of neuroscience, a general organizational principle, one that is conserved throughout life on Earth, has yet to be discovered. There are simply too many fundamental questions about the brain that have not been answered. Many aspects of terrible diseases and disorders, such as Alzehiemers and schizophrenia, are poorly understood. As scientists, there is almost an obligation to answer these questions – an obligation that necessitates the use of animals. In summation, Dr. Mooney explained that the most important quality researchers must have in animal research is mindfulness. We must always consider the implications and gravity of using animals in our studies. What we must be wary of is dismissiveness – the lack of thought or consideration about the privilege that comes with being able to conduct this form of research.

Dopaminergic Neurons: That’s Pretty Dope

First full week at the Mooney Lab: complete. What can I say, this week has assuredly presented some steep learning curves. From accidentally sacking my bird (an affectionate euphemism I suppose) during a brain injection surgery to being unable to find the left ventricle during a perfusion (the process of draining the blood from a sacked bird), this week has posed many challenges. Nevertheless, learning all of these techniques and being thrown into the deep end of neurobiology research is quite exhilarating. I constantly find myself thinking, “Woah, this is sooo cool! (for lack of better words haha).” 

In the short time I have spent with the lab, I am by no means an expert on the project I am about to conduct; however, I will try my best to explain it here. Yet, before I do that, I think it will be helpful to define a few terms (learning these definitely helped me in understanding my project):

dopaminergic neuron cells: a class of neurons that synthesize the molecule dopamine

tyrosine hydroxylase (TH): enzyme that is essential in the synthesis of dopamine; all dopaminergic cells will have the TH enzyme

VTA/SNc: areas located in the midbrain of the zebra finch that are noted by a high concentration of dopaminergic cells

green fluorescent protein (GFP): protein that fluoresces green when exposed to certain wavelengths of light; commonly used as a reporter of gene expression

For my project, I will be testing whether a new cell editing technology is functional and expressed in the dopaminergic neuron cells of the zebra finch. For proprietary reasons, I can’t say the name or describe exactly how this new technology works. However, at a surface level, the technology is able to recognize and bind to a chosen RNA sequence within a cell (in my specific case, it is the sequence that encodes the TH enzyme for dopaminergic neurons). The technology also contains the sequence for GFP. If the technology successfully binds to the TH sequence, then GFP should be expressed. To test this, I will surgically inject the technology into the VTA/SNc region of the zebra finch and “let it sit” for around a month. After this waiting period, I will sack the bird and analyze the dopaminergic neurons within the VTA/SNc and determine if the technology was successful. 

So, in a nutshell, that’s my project! Right now, I am familiarizing myself and learning how to conduct all of the necessary techniques. So, I still have a little time before I actually start the experiment. For now, it’s just learning surgical procedures and how to do bird haircuts (yes, it is exactly what it sounds like).

Nothing But Birdbrains

I think all of us know someone who we would describe as a birdbrain. You know, that one friend or distant cousin who’s just not completely there all the time. I’m not quite sure how this saying came to be, but regardless, we may have to reconsider our use of this term. Turns out, birdbrains are quite complex – go figure – and share many similarities to the human brain. Song birds, and in my specific case zebra finches, are one of the only non-human species who learn to vocalize, and are thus an ideal model to help elucidate the mechanisms that control vocal learning in humans. This summer, I’ll be devoted to studying the neural mechanisms and brains of zebra finches with the Mooney Lab. 

I’ve only been in-lab a few times so far, but here are a few key takeaways: 1). Make sure to close the doors when handling the birds (they WILL try to escape) 2). The brain is remarkably complex. Referring to that second point – while attempting to understand the intricacies of the brain can be quite daunting, it is something that I expect from this summer. I expect to be confused – thrown into the deep end of neurobiology and scientific jargon. Yet, I don’t believe this to be a negative thing. Confusion leads to questions, and questions serve as the foundation for new discovery and innovation. Thus, while I expect to be confused, maybe even overwhelmed by the breadth of new material presented before me, I also expect myself to start asking questions – questions that have answers to them and possibly questions that have yet to be asked. 

Finally, I expect and am very excited to form new relationships this summer. The opportunity to be mentored by and work alongside individuals who are at the forefront of innovation and science is something I do not take for granted. I’m truly thrilled to meet and forge connections with faculty and fellow students who are so passionate about their research, and I am hopeful that some of their passion influences my work as a scientist and individual.