Wisdom from the Giants

From Nobel Prize Laureate Dr. Robert J. Lefkowitz to Mohammad Noor, winner of the Darwin-Wallace Medal, we have certainly had our share of notable speakers. I enjoyed hearing from the distinguished members of Duke’s faculty immensely during my time as a Howard Hughes Research Fellow. Sadly, my summer fellowship is nearing its end. But as we wind down with one more final week of research, I’ve had the time to reflect on the amazing talks I've heard from some very different and very inspirational speakers.

We heard about extremophiles from blooming new professor Amy Schmid and iron homeostasis from the distinguished Dean of the Duke University School of Medicine Nancy Andrews.  Although the subject of their research varied immensely, there were quite a few things that all of the speakers shared. I think it’s important to note these qualities, as they are arguably what makes one a successful scientist. These speakers are some of the most prominent and successful scientists in their field.  They are the epitome of years of hard work. So I were to tell you what all of these great figures share, I’d say it’s their distinguished curiosity, their intellectual drive, and their extreme perseverance. As a scientist, you may study the same topic, trying to figure out the tiniest details of something that most people haven't the slightest awareness of. But you must have faith in your research, and you must work hard. You must persevere.

As I said in my previous blog, science is a sun shower, “ Sure we hear about amazing technology and scientific discoveries all year round, such as the Higgs Boson, water-powered jetpacks, robots, etc. but all of these things took YEARS of hard work, and many, many, mess-ups.” As such, it takes a certain type of person to persevere through, and find the forever-prized rainbow.   

Out of all of the seminars, Huntington Willard’s stood out to me especially. Not only was his research genetics and chromosome activation very interesting, but his guidance and words of wisdom after years of successful research I thought were quite inspiring. Luckily, I managed to get this entire great list down.

Good luck in all of your own endeavors. If one of those endeavors includes a career in science, then may this list be of help to you!

Sun Showers

Research is like a sun shower. For those of you who aren’t familiar with the term, a sun shower is just as its name describes-rain while the sun shines. The phenomenon is unusual and rare to say the least. People are often captivated by their obscurity. It seems odd to compare science to sun showers, but I can tell you that they are actually quite similar.

I’d like to refer to something I said in one of my previous blogs, “As scientists this summer there will be times that we fail, times that we mess up, when our experiments don’t go as we anticipate.” After six weeks working in the lab this summer I am reminded how true that statement is. When we learn about physics, biology, and chemistry what we learn is sound and logical. We take this modern knowledge for granted. There was a time when we did not know what DNA, cells, gravity, or atoms were.  Sometimes theories of the past sound ridiculously silly to us, and the knowledge we now know seems like the obvious reasoning. But science isn't as easy as it looks. There’s a reason why no one figured out the laws of physics for thousands of years, and that reason is that science is hard.

Sure we hear about amazing technology and scientific discoveries all year round, such as the Higgs Boson, water-powered jetpacks, robots, etc. but all of these things took YEARS of hard work, and many, many mess-ups. That’s where science's similarities to a sun shower come in. Like the rain, science is often filled with obscurity, frustration, tired late nights, mess ups and confusion. At the same time however, the sun shines. There are days when your research goes well, when you discover something great, or when you simply enjoy engaging in your research. Though the rain may be coming down, it’s a gentle one. There are of course occasional storms. But, the point is that the sun is still shining! Even when you are burdened with a difficult protocol or failed experiments, all of your efforts are important. The day you fail a protocol is the day you practice and shape your skills so that the next time you perform it you have learned, and have gained expertise. The day your experiment didn't go as planned may be the day you discover something new, and yes, the hours of hard work will pay off, trust me.

So no, science is not a walk in the park. It’s not a perfect sun shining day. It’s a sun shower, which is followed by the occasional rainbow.  

Germinating Success

As part of the Howard Hughes Research Fellows Program we have the chance to hear about our peers’ research through “chalk talks.”  A chalk talk is essentially a presentation that involves no posters or powerpoints, simply a piece of chalk (or a marker in our case) and a board to write on. 

Although we are in a program together, and the same foundation is funding our stay here at Duke, all of our work is independent. Each of us works in separate labs doing very different research. That’s where the chalk talks come in. It gives us the chance to hear what our colleagues are doing. Though these brief talks are just a taste of the incredible work the fellows are doing, it’s amazing to hear about some of the great work being done.

I’d like to talk about Lien Hoang’s Research on the model organism Arabidopsis. Arabidopsis is a weed that grows in North America, her plant –based genetics research is very different from In particular, we seek to better understand how the conditions experienced by the mother plant influence patterns of gene expression, and consequently the germination phenotypes of the seeds the neurological research I do at the Silver lab. It provides an interesting contrast to the work I am familiar with. Therefore I’ve decided to talk about Lien’s research in this blog.

The Arabidopsis plant:

Lien works in Dr. Kathleen Donohue’s lab. The lab uses the plant to study genetics. As Lien writes,” In particular, we seek to better understand how the conditions experienced by the mother plant influence patterns of gene expression, and consequently the germination phenotypes of the seeds.”

According to Lien, there are 3 main factors that influence germination, genes, environmental factors, and maternal effects.  Below is a chart I’ve created that simplifies these factors.

These factors influence germination in a number of ways. Genes influence germination as the genetic makeup of seeds can influence germination timing. Specific genes can control whether a plant germinates at a specific temperature, for example. Environmental factors influence the phenotype of seeds. Temperatures that are too hot or too cold for a plant can prevent or slow down germination. Additionally, water potential, or water’s tendency to move from one location to another, can also have an impact on phenotype, and consequently germination timing. Finally, maternal effects can significantly influence germination. This factor is by far the most difficult to analyze, as it is more indirect than the previous two. However, it is well established that the conditions experienced by the maternal plant can greatly influence the germination time of its seeds.

Studying the influence of the maternal plant can be quite useful. After all, conditions experienced by mothers during pregnancy can impact her child. Smoking and alcohol are examples. Thus, these plants are a useful tool in helping to build our understanding of maternal impact.

Lien was certainly an expert in her field. She knew details down to the most precise point. If there’s a general theme I learned from these chalk talks it’s that we are all blooming scientists. With dramatically different projects, that are often extremely complex it’s great to see myself and my peers developing into scientists. Though most of us are simply at the roots, we have just begun the process of becoming successful scientists. I hope that all of us will continue to germinate, growing a successful scientific career in the process. 

Sincerely Yours,

Danielle 

A Day in the Silver Lab

Working in a lab isn’t like most other jobs. There isn’t a set schedule of events, or a routine that you do every day. Every day in the lab is very different. It all depends on the experiments you are currently conducting and their results.  Of course there are quite a few techniques and protocols that I perform quite frequently.

I start out my day at 7:30 am.  After I get dressed and ready I head out from my central campus apartment lab-bound. Depending on the bus’ timing (which is frequently delayed due to on-campus construction) I either walk the full 30 minute trip, or I take the bus 2/3s of the way. I get to the lab between 9- 9:30am. On Monday mornings we usually have lab meeting at 9:30am, and usually once or twice throughout the week my lab and I go to some of the department’s talks. Although my lab is extremely interdisciplinary we are a part of the Department of Molecular Genetics and Microbiology. 

If I don’t have meetings to attend, well then it’s time for science. Since my routine varies on a daily basis, I thought it would be best to give you an idea of some of the techniques and protocols I encounter most frequently. Often in a lab you will perform the same set of techniques multiple times a week. In my lab these are in utero electroporation, immunofluorescence and FISH.  In this blog I’m going to describe in utero electroporation. Other common techniques include PCR, gel electrophoresis, western blots, southern blots, and transforming plasmids.

In utero electroporation is one of the most difficult techniques. The science behind it is fairly simple. The real difficulty comes from the precision and dexterity necessary to perform the procedure. Let’s start with the phrase “in utero.” In utero is a Latin phrase that can be translated to “in the womb.” When this term is used in biology, it refers to a procedure that is performed on an embryo or fetus.  Electroporation is a technique used in biology. It is when electrical pulses are applied to allow substances through the membranes of cells by making the membranes temporarily porous. These substances frequently include foreign DNA that would like to be introduced into the cells. In utero electroporation is simply exactly as its title describes, electroporation “in utero,” in an embryo.  Below is a diagram showing the process of electroporation on a mouse.  I use electroporated brains in nearly all my experiments. However learning to electroporate is very time consuming and difficult, so at the moment I use sections electroporated by my secondary mentor.

Working in a lab requires more than just individual intelligence and skill. At the core of a successful lab is teamwork. Lab members rely on each other to teach new techniques, answer questions, provide helpful insight and information, and for guidance. Collaboration in the lab is essential. After all two heads (or three heads, four heads, ten heads, and even twenty heads depending on the size of the lab) are always better than one! My daily life in the lab involves working with my wonderful labmates.

Meet our team!

My intelligent secondary mentor Louis-Jan! His French accent is surprisingly faint! 

 My primary mentor and the principal investigator of my lab, Dr. Deborah Silver!

 My buddy in the “mitosis corner” Emily. Her quirky humor provides the occasional laugh.

 Helen is my Chinese speaking buddy!

Lomax does some amazing research on the genetics of brain evolution. He is a helpful hand if I have a question.

    

Though I don’t have pictures, there is also Ashley, who taught me how to use the cryostat, and Samuel a rotating graduate student.

Sincerely Yours,

Danielle 

Understanding the Developing Brain:Neurogenesis and Corticogenesis

The first few weeks in the lab I find are the most difficult. You’re in a new place, and you're doing complicated science that you likely have no experience with. To make it even more intimidating, most members in the lab are skilled, intelligent, and extremely knowledgeable of not only the lab’s research but a plethora of subjects. In my lab, we study mitosis and differentiation of neural stem cells. Our research roughly falls into the fields of molecular genetics, microbiology, developmental and stem cell biology, and of course, neuroscience. In other words, my lab is extremely interdisciplinary.

But as I spend more and more time here I am familiarizing myself with the techniques, tools, and basics of working in a neural stem cell lab. If I wanted to sum up my research in one statement I would tell you that I am doing an anatomical study of the basal process of neural stem cells during mammalian corticogenesis. But to someone who has no background knowledge of neuroscience and molecular biology this description has very little meaning. So I’m going to start with corticogenesis.

Corticogenesis is the formation of the cerebral cortex, or the outer lining of the cerebrum. The cerebral cortex, which is also commonly referred to as the neocortex in mammals, is made of six layers which vary in neuron concentration and function. The development of this complex structure, also known as arealization, is consequently a highly specific process whose formation is largely reliant on spatio-temporal timing.

During corticogensis, neural stem cells (NSCs) also known as radial glial cells (RGCs) go through a process known as neurogenesis in which mature “adult” neurons are generated. The following figure displays a simplified version of this process:

However, as shown in the figure, this pathway is not always direct. NSCs can directly differentiation into mature neurons, or they can first differentiate into basal progenitors (BPs), also known as intermediate progenitor cells (IPCs), and then later, following another round of mitosis, differentiate into mature neurons. If NSCs do not differentiate following mitosis, this is known as proliferation. Both daughter cells will be NSCs.

The following diagram displays the direct and indirect forms of neurogenesis. It is important to note that when NSCs differentiate following mitosis, one of the daughter cells remains a NSC while the other may be either a basal progenitor or a neuron. Never do both daughter cells differentiate, except when an IPC undergoes mitosis to form two mature neurons.

My research focuses on understanding neurogenesis and the several factors that contribute to neuron differentiation. One of these factors, regulation of the cell cycle, specifically mitosis, is critical for cortical development. After all, it is the process of mitosis which produces mature neurons. Understanding the mechanisms underlying the division of these stem cells, or radial glial cells is crucial. These NSCs are long neuroepithelial cells which stretch all the way from the ventricle zone (VZ) that borders the ventricles of mammalian brains, to the pial surface of the brain. To reach such great lengths, these cells have a long epithelial structure known as a basal process. At the end of the basal process are structures known as “basal endfeet.”

It has been found that cell polarity places a crucial role in neurogenesis. There is strong evidence that supports that daughter cell inheritance of the basal or apical process following mitosis plays a crucial role in the determining of cell fate. This summer I am trying to understand  the importance of the basal process in neurogenesis. Using techniques such as immunohistochemistry, live imaging, and in utero electroporation (thanks to the assistance of my secondary mentor Louis-Jan!) on brain slices I am trying to identify some of the factors that influence cell fate and the mechanism by which they accomplish this.

We believe that RNAs localized at the basal process may play a critical role in neurogenesis. Using immunoflorescent antibodies we can label the nuclei, cell body, basal processes, and specific proteins within cells. Below is an example of some of the immunofluorescence we do here in the lab.

mCherry is a fluorophore that we use to label the cell body and basal process. HOECHST labels the nuclei, and FMRP targets mRNAs. FMR1 is the human gene that codes for the protein FMRP, which is critical for cortical development. Mutations in the FMR1 gene can lead to a variety of mental and physical disorders such as mental retardation, autism, fragile x syndrome, and parkinsons. It is just one example of the many proteins essential for cortical development. Hopefully I can contribute to our knowledge of neurogenesis so that we may better understand the mechanisms of brain development.

Learning from the Master

This summer I’ll be spending my time studying brain development in the Silver Lab. The research I’ve been doing here these past two weeks is groundbreaking. It’s been a dream of mine to work in a neuroscience laboratory for some time now. I am specifically interested however in developmental biology. With its cutting-edge neural stem cell research, the Silver Lab manages to combine two of my greatest interests, developmental biology and neuroscience.

Having just started out here, I have a lot to learn. Though I have worked in other labs previously, I find that laboratories vary with surprising specificity. Although some of skills from working in my other lab have helped me greatly, there are many techniques I have yet to master. I have tried, several times to isolate completely intact brains from developing mouse embryos, and have yet to master the art. Of course sharpening my skills and developing expertise in research requires time and often mentorship. I have the pleasure of working in a welcoming and encouraging lab. What better way to learn the ways of research than to interview a master?

I’d like to introduce my primary mentor, Dr. Debra Silver an accomplished scientist and the principal investigator of my lab. In the following interview Dr. Silver or Debby for short, reveals to me her journey and experiences in science.

Here is a photo of  my secondary mentor Louis-jan (LJ) and me in the lab!

Where are you from?

I grew up in Massachusetts in a small town called Southboro, which is 40 minutes west of Boston.

Were you always interested in science? 

Yes. I think I realized I loved science in high school. I wasn’t one of those people who grew up doing science experiments here and there. I took my first biology and chemistry courses in high school, and I went home and realized I liked doing my chemistry homework every day, well, that’s when I realized I liked science.
 

Where did they go to school and why did u go there? What was your major?

I went to Tufts in Massachusetts. I first became interested in Tufts because my father went there. I decided to go there because I loved their campus and I knew that they had a pretty strong program in biology and I wanted to be a biology major. I also wanted to be closer to Boston.

How long have you know that you wanted to pursue a career in science and specifically neuroscience?

When I was in college I started doing research for three summers. After I graduated I decided that I wanted to do more research and so I went back to the same lab that I did summer research in. I ended up staying there for four years. I originally had planned on going there for two years, to save enough money to travel around the world. I got really into my project, and my boss at the time treated me more like a graduate student than a technician. I was a lab technician at the time. I got so into it that I ended up staying for four years and I got a couple papers out. That convinced me that I wanted to go to graduate school. I applied after I had been in the lab for two years and I deferred for a year. I had one first author paper and two coauthors as a lab technician. That was how I got excited. I got interested in Neuroscience towards the end of graduate school. In graduate school I had been working on basic science questions on how cells migrate in drosophila. I decided that I wanted to move to a system that had more relevance to humans, so that was mice. I was particularly interested in the nervous system because it’s so fundamental for life.

What do you like best about being a researcher?

I love the thrill of discovering something that no one else has discovered before and I love the opportunity to peer into biology and learn about something that’s been perfected by a cell and chip away at our understanding of it. I like the creativity involved in coming up with new experiments and the thrill of finding something new.

Any disasters in the lab or embarrassing moments?

When I was a summer student I had spent a week making a radioactive probe. I lost sight of time when I was boiling it and completely melted it all over the heat block. I also found out the hard way that xylene has to be stored in glass and not plastic. It melts plastic. I did that experiment inadvertently and it melted a whole thing of plastic.
 

Have you had experience in a non-academic environment?

Well I did my postdoc in a government lab. That was a very different experience. I’ve learned that I love academics. I think it’s a huge part of being a scientist to teach other people what we know. I think the science can go in really exciting directions when people who are working on the problem have no bias as to what they think is going to happen. The government lab was a very exciting place to work. The world is your oyster as a scientist, you have so many resources.

What do you like to do outside of science?

I have two kids so most of my free time goes towards them. I love mountain biking. I like photography. I like mostly just spending time with my family, playing soccer with my kids. I like yoga too.

How did you end up at Duke?

When you are looking for a job you apply to a bunch of places that have labs. I ended up deciding on Duke because it was a great place that combined my interested in neurobiology, developmental biology, and RNA biology and there are a lot of people doing that here. I have pretty broad interests.

How has your experience been teaching?

I really like teaching. For a long time I thought I wanted a position that was more teaching and less research. I wish teaching was more appreciated. I love teaching and having people in my lab. I guess that’s more mentoring. I love when people take a class out of interest and not a requirement. I’ve always tried to teach. Actually when I was a graduate student I tried teaching students in high school in Baltimore.

What is your greatest finding?

I don’t know what my greatest finding is. But my most exciting finding was in graduate school. Hopefully everyone has that day when they find something really exciting in the lab. For me, that was in graduate school. I had generated mutations in flies in the components of the JAT/STAT signaling pathway. I had discovered that this pathway was required for cell migration in fly ovaries. My most exciting experiment in graduate school was the day that I overexpressed components of this pathway in post-mitotic cells. This caused a huge increase in the number of cells that migrated. This showed that JAK/STAT was not only required for migration but was also sufficient to tell a cell to migrate. I was third year graduate student and it was a very unique finding. That was the first really big finding of my career.

What are you looking forward to most?

Having our lab really make an impact on the field of stem cells and neural development, cortical development, and all of the members of our having the chance to taste success and make impacts on the field. I would also like to potentially identify things that have an important influence on neurodevelopmental diseases.
 

What would you change about doing science?

I wish the public appreciated the time it takes to do science and the importance of basic science towards finding cures for disease. I wish it was more appreciated.

What lessons do you have or advice for someone pursuing a career in science?

I think find out what you are passionate about and work on it. I think it’s really helpful to find a mentor, someone you can look too as a role model, and be patient because science requires a lot of patience. But if you put in the patience it will reap awards back at you. Any career, science or otherwise, is just that you are passionate about it. Graduate school is a lot of work, be passionate about it. If you are looking for opportunities in labs, look for places that are nurturing and helping you develop as a scientist as opposed to just treating you as a person who is there. You have to be willing to put in the hard work. I think it’s important to always maintain enthusiasm.

Build Your Own Door

Let me introduce myself. My name is Danielle Scarano. I'm an 18 year old rising sophomore at Duke University. I'm currently working in a neural stem cell lab  If you were to ask me who I am there are many different replies I could give. I’m an illustrator of imagination, and a master in the art of sarcasm. I’m a onetime skier, an expert taco maker, and an theoretical physics documentaries addict. I’m an atrocious singer, and an incompetent swimmer. I stand at a mere five foot one, have green eyes, and a terrible fear of spiders. I am a gold medalist in the International Sustainable World Project Olympiad, a two-time All County Artist, a Simons Research Fellow, a Siemens Semifinalist and a finalist in the Hong Kong International Science fair. I could tell you that science has always been my passion, and that I could recite the planets backwards and forwards at age three.  I could tell you many things, but alone these things do little to define me. So come along with me on my journey, as I try to figure out who I am and what I wish to pursue. 

There’s a little quote that I’m quite fond of by William J. H. Boetcker. It goes, “The difficulties and struggles of today are but the price we must pay for the accomplishments and victories of tomorrow.” In order to effectively articulate my expectations for this summer, I think it’s best to tell you a little bit about how I see things. Everyone looks at the world from a different perspective. Everyone has faced troubles, terrors and triumphs. I would say that all the Howard Hughes Research Fellows have something to be proud of. We are at one of the best institutions on the planet, especially when it comes to cutting-edge research, and all of us hit the ground running. Duke is a realm of endless opportunity, and we took it. I think that alone is exceptionally commendable. There are those who see opportunity and are too fearful to go for the gold, and there are those who take it. Life is too short. We must be bold in our pursuits. And even if opportunity doesn’t present itself to us, we shan’t let that hinder our progress. Persistence is something of incredible value. It is what distinguishes those who reach for the stars from those that actually grasp them.

As Milton Berle once stated, “If opportunity doesn't knock, build a door.” This is the philosophy I follow, or at least try my best to. To demonstrate this best I’d like to quote something from my Howard Hughes Application, “In science it is inevitable that we will sometimes fail. As we test our hypotheses and attempt to answer questions we are often wrong. Yet if our drive to find answers is crushed by a mere failed attempt than science could never be successful. If the great minds of the past had given up after their initial hypotheses failed, then science would never have progressed to level it is at today.” That is why inventors, businessmen, and most of all scientists must build doors. They must formulate alternative solutions, deduce answers to questions, and frequently oppose a widely accepted idea. They must enlighten and challenge.

So I say, today’s struggles are tomorrow’s successes. As scientists this summer there will be times that we fail, times that we mess up, when our experiments don’t go as we anticipate. Although I have already worked in several labs, I expect that I will have some failures. Even the best scientists do. Some of the most interesting and revolutionary discoveries have resulted from mishaps. Perhaps then it is wrong to describe these mistakes as “failures.” It is probably more accurate to label them as discoveries.

As Thomas Edison once said, “I will not say I failed 1000 times, I will say that I discovered there are 1000 ways to cause a failure.” This summer I expect to grow as a scientist. I am now at a point in my life where I must decide which direction I want my life to go. Although I have known since I was young that it was in science, it is only through the anticipated practice, persistence, and discovery that my time as Howard Hughes Research Fellow will provide that I will be able to develop a clearer understanding of who I am and what doors I wish to create.

Me working in the lab!

Hopefully my corny motivational speech has inspired you to be bold in your own pursuits.

Sincerely Yours,

Danielle