Author Archives: Kacia Anderson

The End But Not Really The End

This summer I was fortunate enough to spend eight weeks conducting research in the Bejsovec Lab. In addition to this research, I had opportunities both explore Durham and learn from faculty as they shared their research journeys with us. Beyond learning about these journeys, BSURF allowed me to think deeply and critically about how I want to spend my remaining years at Duke and the type of person who I wish to become by the end of my undergraduate career.

I believe my summer experience influenced my perceptions about research in a positive way. Before this summer experience, I never had a clear idea of what the life of a researcher looks like, how slow science could be, but at the same time how rewarding failures and successes can be. I believe among the many experiences I’ve had these past eight weeks, the people that I’ve met and the memories created are the best parts of this summer in addition to learning how to conduct research and how to communicate science.

This summer has allowed me to become more certain of the career I would like to pursue in addition to showing me the multitude of routes that I can take to achieve my ultimate end goal. Furthermore, the advice given to us throughout the past eight weeks has not only made me wiser, but certain that the end of BSURF isn’t truly an end.

I would like to thank BSURF and the Bejsovec Lab for providing me an amazing opportunity to learn and observe science, as well as my fellow BSURF-ers for an amazing summer experience.

Don’t Just Take Advice, Evaluate It

As a component of my summer research experience, I have been very fortunate to hear from faculty researchers from diverse fields. Many of these sessions were filled with learning about not only the research that these faculty members do here at Duke but also how they came to find themselves doing research at Duke, and what opportunities research has afforded them. Personally, I found it very interesting to hear about the somewhat linear and more often then not so linear paths that led them to doing what they love.

One talk that stood out to me in particular was Dr. Mohamed Noor’s presentation on his meanderings on evolution and species formation. The research Dr. Noor conducts is focused on biological evolution that occurs within populations of Drosophila as a result of variances in recombination. More so how these changes in genetic recombination overtime lead to the formation of a new lineage of a species. I really enjoyed learning how phenomenons like antibiotic resistance which pose great threats in the post antibiotic era is a great demonstration of natural selection occurring within our environment and can have real consequences for humans in the long term.

In addition to learning more about his research, Dr. Noor left us with a few pieces of advice that deeply resonated. This advice from his own journey to become a researcher are to take chances and initiative, evaluate the advice given to you, and also, do something you love. As I continue in my undergraduate and research career, I hope to embrace this advice in the future decisions and directions I may pursue.

 

 

Abstract

In Drosophila Melanogaster (fruit flies), wingless(wg) a Wnt growth factor responsible for cell to cell communication operates in two parts, a signal transduction and movement (of this signal) to other neighboring cells. What remains unclear to researchers who study wingless is how wingless protein is moved and the molecules that help move it. The Bejsovec Lab’s hypothesis is that molecules that aid the movement of mutant Wg protein will be able to influence the wild type Wg protein. In order to test this hypothesis, cuticle preparations of mutant for wingless and these suppressor genes were scored for improved cuticle patterning in double mutant embryos. In order to confirm that the changes in cuticle pattern were due to increased movement of the wgNE2 protein, antibody staining was performed at early developmental stages. The findings of this project may uncover that these molecules are able to aid not only mutant wingless but wild type as well.

What to Expect When You’re Expecting

So this week’s blog post is centered around my day to day experiences in the lab! I’ve had the title for this blog post since the day 1 meeting when Dr. Grunwald suggest we use interesting, creative titles for our blog posts. Anyways, on to explaining what I do in lab most days.

Usually, I start my days in lab around 10AM after the morning session with either faculty in the Biological Sciences or an informational session given by Dr. Grunwald or Anna.

The first thing I do when I get to lab is check on my morning fly check. This consists of me getting my fly babies (if you cannot tell I am very fond of my flies) that are in the incubator out and examining the vials looking for empty pupae cases. We keep flies in the incubator (25 degrees celsius) because it helps slightly speed up the development of the flies without causing any disruptions to development or manipulating any temperature sensitive mutations.

After checking on my crosses, I either transfer the adults from a cross into a fresh vial with new food to expand the cross or I will look for male and virgin female flies in order to set up a new cross. To look for flies, I first have to turn on LED light on the microscope and the CO2 gas which is filtered through water and to this small nozzle (I can control the release of gas through this nozzle manually). I can then shoot CO2 gas into the fly vial (I hold the vial upside down during this process –it keeps the flies from drowning in their food) which puts them to sleep. I then remove the cotton bung which plugs the vial and slightly tap the vial to get all the sleeping flies onto a pad. This pad (I like to think of it like a bed) is made of a porous material and is also attached to the hydrated CO2 causing the CO2 to be pumped onto the surface of the pad keeping the flies asleep while I sort through them using a paintbrush under the microscope. After I collect the flies I need to set up a cross, the other flies get funneled in the fly morgue (RIP fly babies).

Additionally during this check, I will also transfer freshly eclosed flies into a new vial if I am building a stock (flies that are all isogenic) in order to expand the stock (as my PI says, it’s always good to have a back up). After I am finished checking on my crosses/setting up new crosses, I return the flies back to the incubator so that they can continue aging. I repeat these same steps in the afternoon before I leave for the day as a part of my afternoon fly check.

After my morning fly check, I usually have a few options as to how to spend my day until I perform my afternoon fly check. The first week fo BSURF, I spent this time reading different fly protocols and literature about Drosophila. I also took time to understanding more about the purpose of the crosses I will be setting up and the different phenotypes that will result from a particular cross. My PI also uses this time to teach me a new technique that I will need for something I will be doing later in the day. There have also been points during BSURF where I set up blocks to collect eggs from specific lines of flies to mount on slides which would usually take me about 2 hours depending on how many block positions I had filled.

Currently, I spend the time between my fly checks either working on my blog post (which I am currently doing), making mockups of my presentation poster, or thinking/ researching about the next steps in my experiment given the questions my PI brings up for me to think about.

In the afternoon, I perform my fly check mainly to see if there are any freshly eclosed females  of a particular genotype that I need to perform a cross. This is mainly because there is a window of about 8 hours that female fruit flies need to become sexually mature. Before this time frame passes, the female is too soft for a male to mate with thus the female flies I collect during this time have not mated. It is important for me to mate  female virgins because that ensures that I know the possible genetic and phenotypic outcomes of the eggs she will lay since. If I pick an older female, I won’t know for certain the genetic combination of the eggs she lays.

Here’s a picture of the microscope and the CO2 bed

Mutant Bacteria… Can They Do It Better?

I really enjoyed the chalk talks this week and having the opportunity to present a snippet of my own research as well as learn more about my peers research. Of all the amazing presentations, one that really fascinated me was the research of Ella Gunady.

Ella is working in a Bass Connections Lab with Dr. Jason Somarelli on creating a mutant bacteria that can efficiently consume plastic. Aside from doing exciting work with real world applications that change how waste remains in our environment, I found the setup of Ella’s talk very easy to understand. Despite working in an entirely different field and realm form my own research, I was easily able to follow along with the explanations Ella gave us due to the sufficient background and great illustrations that she used.

In detail, Ella will be creating a library of mutant bacteria that can consume plastics and micro-plastics through an application of error-prone PCR on the bacteria that already degrades plastics naturally. After creating this library, Ella will be testing the efficiency of each of these mutants through exposure different environment compositions to identify the best candidate. The findings of this research will have significant implications for the environment, especially in communities that have high levels of waste.

In all, I cannot wait to see how all of my peers research translates into interesting   presentations as the end of the program!

At the Edge of Knowledge…Getting to Know Dr. Amy Bejsovec!

I had the pleasure to sit down with my PI, Dr. Amy Bejsovec and her dog Benny, this week to interview her for my blog post. I was really excited for this opportunity to learn more about her and her research journey.

Dr. Amy Bejsovec started out at Cornell University as a biology major. She then continued her education at University of Wisconsin Madison after being accepted into their genetics program. However, her love of science started way before her undergraduate major. Growing up in near a wooded area on Long Island, Dr. Bejsovec spent much time in nature venturing out as far as her bike could take her. She told me that she found herself just fascinated with animals and wanting to learn more.

With such a wide area of interest for nature, I wanted to know more about how Dr. Bejsovec came to be working with Drosophila melanogaster. Dr. Bejsovec did not always know that she would work with Drosophila, in fact her first encounters with an organism occurred through a project on territorialism in hermit crabs that won her first prize (a calculator that she still has and occasionally uses till this day!). Throughout graduate school, Dr. Bejsovec worked on muscle development in nematode worms. When Dr. Bejsovec was deciding what to do for her post-doctoral work, new discoveries regarding the striped nature of gene expression in Drosophila caught her attention. From then on, Dr. Bejsovec has spent much of her career researching wingless.

A fascinating moment in her research journey she told me about occurred during her third year of graduate school when she truly recognized herself as a scientist. Up until that point, she thought that these unanswered questions were a shortcoming of her own knowledge; however, she came to realize that these questions signified the edge of knowledge. But after recognizing this boundary that she was approaching; she decided to persevere and push forward (a common theme in her research journey) to begin finding the answers to the questions.

In her own lab at Duke, she continues working with winglessand Drosophila and foresees herself continuing her work with this model. Outside of her research Dr. Bejsovec co-teaches Biology 202 Genetics and Evolution, and a class on Cancer Genetics. In addition to teaching roles, she serves on the Academic Council for Duke University. Towards the end of our interview session, I asked Dr. Bejsovec about what advice she would give to someone wanting to get involved with research. Though our conversation Dr. Bejsovec imported many lessons of persistence and perseverance but I leave you with a few things that resonated deeply with me:

 

“Find what you’re passionate about, what makes you feel fulfilled and satisfied and pursue that”

“Talk to as many people as you can who are willing to speak with you and don’t just listen to one person.”

“When you recognize that you’re at the edge of knowledge… the question is how do we push beyond it?”

 

Read more about her and her work on her lab website and Duke informational page.

 

Not Soaring but Definitely Flying

All animals use Wnt growth factors for controlling cell fate decisions as the organism develops. The wingless gene (wg) encodes the Drosophila Wnt growth factor, Wingless (Wg) protein. In Drosophila, a loss of function mutation in the wingless gene causes a disruption in epidermal patterning during embryogenesis. At the end of embryogenesis, a wild type fly larva would have a cuticle pattern, produced by the epidermal cells, that consists of repeating segments that alternate between a belt of denticles and a region of naked cuticle (see image below).

Cuticle patterning on a wild type fly.

A complete loss of function mutation in the wingless gene eliminates the regions of naked cuticle that separate denticle belts, resulting in a continuous area of denticles (see image below). In both flies and humans, Wnt growth factors associate tightly with cell membranes, but they are able to control cell fate decisions at a distance from their origin points. In the fly, some mutations in the wingless gene disrupt the cell to cell movement of the protein without altering its signaling. For example, the wgNE2 mutation restricts the range of protein movement, and so is not able to control cell fates at the normal distance seen in the wild type.

Region of denticles with no naked cuticle to create proper segmentation.

The Bejsovec lab has identified two suppressors that improve the cuticle patterning of the wgNE2 mutant embryos. For my project I will be working with these suppressors to characterize how they impact the movement of the WgNE2 protein and to test whether they affect movement of the wild-type Wg protein. As a part of my project I will be doing antibody staining for the WgNE2 protein and viewing my preps using a confocal (fluorescent) microscope.

A hope that stems from my project is that these same suppressors will also have impacts on the movement of wild type Wg protein. The discovery of movements in the wild type Wingless protein could possibly lead to groundbreaking discoveries given that the Wingless growth factor is very similar to the Wnt1 growth factor in humans, which is required to pattern the central nervous system. In mice, knocking out the Wnt1 gene results in lack of development of the cerebellum, and death of the mutant embryos (to read more about this, click hereUnderstanding how Wnt proteins move will help us understand how our nervous system is patterned during development. 

A special thanks to my principal investigator, Dr. Bejsovec, for helping me in this blog post.

Credit for the images and information below.

Dierick, H. A. & Bejsovec, A. Functional analysis of Wingless reveals a link     between intercellular ligand transport and dorsal-cell-specific signaling. 10

Drosophila, the Lab, and Me

This past Monday I officially started in the Bejsovec Lab, a biology lab that studies molecular mechanisms for embryo development in the Drosophila or fruit fly model. After my first week in lab, I am eagerly anticipating the arrival of the progeny from my first fly crosses, which are expected to eclose (or hatch) tomorrow! This first week has definitely been filled with challenges as I grapple with understanding my project and how to think about the expected outcomes from the crosses performed. I have definitely made my fair share of mistakes this week but at the same time I have learned a lot pertaining to my project. I am excited to continue learning and begin (hopefully) making discoveries (big or small) within the scope of my project.

 

My hopes for my summer research experience with BSURF is that I get to learn about my research interests and how I can incorporate research into my remaining years at Duke and beyond. I think up until this point, I’ve had a narrow idea as what my future at Duke could look like. Many people talk about research and seem to know exactly which lab they want to go into and what working in a lab entails. However, I did not. In this first week alone, I’ve learned a lot about the pace of research and how the day to day life in lab can be so different when discussing my day compared to my peers. I anticipate that as the program continues, I will learn more about research in other fields as well as within my own lab’s field.

 

Finally, I hope to make mistakes and learn from these mistakes. As I mentioned previously, I’ve already made many mistakes this week alone. However, as Dr. Grunwald has mentioned to us throughout the week, we will make mistakes and hopefully we will learn from these mistakes to become better scientists. Thankfully, I’ve already owned up to what I don’t know and my gracious mentor and PI, Dr. Amy Bejsovec, has been more than accommodating and willing to explain concepts and ideas to me whenever I have questions. Overall, I am very excited to rise to the challenge that my lab presents me and contribute in any way possible.

Me at my desk in the Bejsovec Lab

I was working on the genotype and phenotype for the crosses I have in the incubator.