First Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping researchers promote themselves alongside their papers. Emily D. McParland and T. Amber Butcher are co-first authors on ‘ The Dilute domain in Canoe is not essential for linking cell junctions to the cytoskeleton but supports morphogenesis robustness’, published in JCS. Emily and Amber conducted the research described in this article while they were postbaccalaureate research assistants in the labs of Dr Mark Peifer (E.D.M. and T.A.B.) and Dr Kevin Slep (T.A.B.) at Chapel Hill Department of Biology, University of North Carolina, USA. Emily is now a PhD student in the lab of Dr Thomas Roberts at Brown University, Providence, USA, where she is broadly interested in functional morphology and muscle biomechanics, and focuses on the function of the collagenous extracellular matrix in muscle force production. Amber is a PhD student in the lab of Dr Eric Greene at Columbia University Department of Biochemistry and Molecular Biophysics, New York, USA, where she uses biochemical and biophysical techniques to investigate protein function, most recently within the context of homologous recombination.

Emily McParland

How would you explain the main findings of your paper in lay terms?

E.D.M. and T.A.B.: During early development of animal embryos, the body plan undergoes dramatic shape changes to morph and differentiate into its larval or adult form. During these movements, cells must maintain a robust linkage between the actomyosin cytoskeleton (which powers cell movement) and cell–cell adherens junctions (which connect neighboring cells together). Our lab investigated the Drosophila Canoe (the homolog of mammalian afadin), a large protein with five highly evolutionarily conserved folded domains. The largest domain in this protein (Dilute domain), shares a similar predicted structure to the Dilute domain of Myosin V. Both Myosin V and Canoe Dilute domains can dimerize. To our surprise, mutants lacking the Dilute domain (CanoeΔDIL) are viable, fertile and exhibit proper protein localization in adherens junctions. However, when we reduce CanoeΔDIL protein expression by half, we see that CanoeΔDIL is no longer fully functional, and there are defects in embryos and developing eyes. Together, these data reveal the robustness of the junction–cytoskeletal connection throughout development and the power of natural selection to maintain protein structures throughout evolution.

Were there any specific challenges associated with this project? If so, how did you overcome them?

T.A.B.: My favorite part of this project was the multidisciplinary approach we were able to take from the molecular scale to the whole animal. However, this approach also posed many challenges. Creating the Dilute domain deletion posed some initial problems as I was not used to work with plasmids of that size in molecular cloning. I reached out to people who had previously undertaken fly work and to old members of Mark's lab, who had created the system I was using, to gain insight into previous challenges and how to overcome them. Next, I completed diagnostic tests in different steps of creating the mutant and was able to come up with an altered protocol for successful cloning and plasmid preparation for mutant fly generation.

When doing the research, did you have a particular result or ‘eureka’ moment that has stuck with you?

E.D.M.: When first carrying out genetic analyses of the CanoeΔDIL mutant, we were seeing confusing results regarding viability. We performed an outcross to remove a potential lethal mutation near our protein of interest. Once we completed eleven generations of the outcross, I reanalyzed the viability of the CanoeΔDIL mutant, looking for homozygous mutants. I remember how thrilled I was when I saw the phenotype that indicated homozygosity and the PCR gel that confirmed it. Once that happened, we were able to carry out further experiments to tell the whole story about the Dilute domain.

Why did you choose Journal of Cell Science for your paper?

E.D.M. and T.A.B.: The heart of the work is an investigation into the cellular events of morphogenesis. We are primarily interested in the integrity of cellular junctions and how they can withstand mechanical force during embryonic morphogenesis. Journal of Cell Science seems like the perfect place to publish our findings, as the journal is committed to furthering the field of cell biology through the publishing of recent novel discoveries. We also want to publish in this journal because it's published by The Company of Biologists, a not-for-profit organization, with a well-established peer review process and a commitment to communicate cellular science with a strong experimental foundation.

Amber Butcher

Comparison of late-stage Drosophila embryos representing wild-type, cnoΔDIL/ΔDIL homozygous and cnoΔDIL/R2 heterozygous phenotypes, respectively.

Comparison of late-stage Drosophila embryos representing wild-type, cnoΔDIL/ΔDIL homozygous and cnoΔDIL/R2 heterozygous phenotypes, respectively.

Have you had any significant mentors who have helped you beyond supervision in the lab? How was their guidance special?

E.D.M.: Dr Mark Peifer, my postbaccalaureate mentor, was fundamental in preparing me for graduate school. He encouraged me to step outside of my comfort zone and explore questions differently from how I was originally trained to do, giving me the confidence to try new things in new places. He will always take the time to help me gain full understanding of a topic and discuss questions I have about our research or professional development.

T.A.B.: My undergraduate mentor Dr Steve Rogers and my postbaccalaureate mentors Dr Kevin Slep and Dr Mark Peifer have all played an integral role in my career development and have continued with their guidance since I left UNC Chapel Hill. They all inspired me to pursue a PhD and exposed me to how exciting and fulfilling lab work and scientific discovery is. From the beginning, they've been invested in my personal success and have trained me to be a good scientist in different fields. They have truly been wonderful to work with. Having great mentors is an integral part of being a young, developing scientist and I am extremely lucky to have been able to be a member of their labs and of the scientific community at Chapel Hill. I have no doubt they will continue to be a part of my scientific journey as I continue to pursue my thesis work here at Columbia.

What motivated you to pursue a career in science, and what have been the most interesting moments on the path that led you to where you are now?

E.D.M.: I first learned that I wanted to pursue a career in science when I took a marine comparative anatomy course at Shoals Marine Lab, where I conducted my own research project while being surrounded by people doing similar work. It was energizing to learn that I could spend my life learning about things that I find interesting. I fell in love with research through my undergraduate experiences where I studied rat chewing kinematics and analyzed the utility of common comparative biomechanical tools in clinical research in jaw joint disorders. After I finished my undergraduate degree, I looked for challenging opportunities in a different field, so I joined the Peifer lab at UNC, where this work originates. I got a chance to ask questions in different ways and gain more experience in developmental biology, which always interested me. I now have since shifted back into comparative biomechanics research and work in a muscle physiology lab for my PhD, and use the different ways of thinking that I learned in the Peifer lab.

T.A.B.: I have always had an underlying love of science and inquiry; however, during my time as an undergraduate, I was able to take two CURE classes (research-intensive courses), which let me delve into the realm of scientific research. Through these courses, I worked on characterizing novel jaw adaptations in pupfish and understanding lipid droplet synthesis in S2 cells. It is also through these courses that I was able to meet Dr Steve Rogers, in whose lab I began working in my senior year. Working in Dr Rogers' lab, hearing his guidance on what a PhD would mean for my career and what a scientific career really looks like, was integral in my motivation to pursue a PhD. My parents were both first-generation college students and are not involved in scientific research. Therefore, the most interesting moments have been meeting mentors who have given me guidance on my academic path and passing the things I learn along to younger scientists, by being involved in the Graduate Student Organization and new student recruitment here at Columbia.

Who are your role models in science? Why?

E.D.M.: Dr Nick Gidmark, my undergraduate mentor, motivated me to pursue a career in research. He carries such a contagious excitement for his research and topics he teaches in his courses. The idea that I could motivate and support students like he does inspires me to pursue a career in research while also teaching about the beauty that biology has to offer. I look forward to sharing his enthusiasm and optimism with my own students in the future.

T.A.B.: My current role models in science are Dr Anum Glasgow and Dr Anna-Lena Steckelberg. They are both faculty in the Department of Biochemistry and Molecular Biology here at Columbia University Irving Medical Center and have started their labs within the last few years. Both are extremely talented scientists, and I admire their involvement with students and the change the department is seeing as new female PIs continue to push the field forward. I have had wonderful experiences with both and look forward to continue to be a part of a department with such exciting research and faculty like Drs Glasgow and Steckelberg. They are invested in not only their own lab members, but also all students, including myself.

What's next for you?

E.D.M.: I am a first-year PhD Student in EEOB at Brown University where I'll spend the next 5 years in the Roberts lab studying muscle physiology and biomechanics. I'm excited to use the skills that I learned in my work in cell biology to help me ask questions through a more-organismal lens.

T.A.B.: Being a second-year graduate student, I will continue to push forward with my PhD, investigating the mechanics of homologous recombinases, using a variety of biochemical and biophysical techniques. I plan to keep being involved in the Graduate Student Organization and other clubs here on campus and also to keep a strong involvement in the scientific community.

Tell us something interesting about yourself that wouldn't be on your CV

E.D.M.: I love to bake cookies and bread for my friends, and I am always ready to start a new craft hobby. Recently, I have been into embroidery and watercolor.

T.A.B.: Aside from science, I really love language learning and music. In college, I also studied Hispanic Literatures and Cultures and I have continued to be enamored with the subject since. There are few occasions where I am not listening to music. Music gives me a little pep throughout the day and lately, 70s soft rock has been my go-to.

Emily McParland’s contact details: Brown University, Department of Ecology, Evolution and Organismal Biology, Box G-W 80 Waterman St, Providence, RI 02912, USA.

T. Amber Butcher's contact details: Columbia University, Department of Biochemistry and Molecular Biophysics, 701 W 168th St, New York, NY 10032, USA.

E-mail: emily_mcparland@brown.edu; tab2186@cumc.columbia.edu

McParland
,
E. D.
,
Butcher
,
T. A.
,
Gurley
,
N. J.
,
Johnson
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R. I.
,
Slep
,
K. C.
and
Peifer
,
M.
(
2024
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The Dilute domain of Canoe is not essential for linking cell junctions to the cytoskeleton but supports morphogenesis robustness
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J. Cell Sci.
137
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jcs261734
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