A Developmental Journey and Lessons Learned Along the Way

A career in science is a journey of wonder and discovery. To succeed in science requires curiosity, perseverance, a good dose of luck, and wise guidance from those who have taken the journey ahead of you. We also need to use our science skills to contribute to public debate on complex issues of the day.Being honored by the 2009 American Society for Cell Biology Women in Cell Biology (WICB) Senior Award has provided me with an opportunity to look back and examine the importance of mentorship and role models, both female and male, in my career. I am fundamentally a basic biologist, driven by my curiosity about how the world works. The question that has fascinated me for over 30 years is one that we can all relate to: How is it that complex, rational organisms such as ourselves can arise from a single cell, the size of a speck of dust?Over the years my lab colleagues and I have explored many aspects of that question, using mice as our model system, and we''ve discovered more and more about the hierarchy of cell decisions that begins when sperm hits egg. Along the way we have contributed to the development of techniques for manipulating the mouse genome, helped identify key signaling pathways that control blood vessel development, and isolated novel stem cells from the mouse blastocyst. But I always return to the fundamental questions of lineage development in the early embryo, attacking the problem with new tools as they become available.     

Looking back on a life of unacknowledged privilege and a call to action

The year 2020 provided a wake-up call about the role systemic racism plays in shaping our nation and shaping science. While hard work and great mentors helped bring me a long way from a farm in Minnesota, it’s become much clearer that the privilege of being white and male and the accumulated advantages that began there played powerful roles. It’s time for white scientists like me to listen, think, and take action.

We all have personal stories that we use to describe our trajectory in life and science. For the past five decades the narrative I told myself was a simple one of good luck, hard work, support from my community, and mentors at pivotal times. However, in many important ways, this was just a small part of the truth, ignoring the role unperceived privilege played. The many underlying injustices that were laid bare in our nation this past year began to open my eyes, prompting me to look back at the roles hidden privilege played in my career and the power that these have given me. This challenged me to use the power of that privilege to speak and act to try to change the system in which engrained advantages benefit some but not all. I am telling my story in hopes it will encourage my white colleagues to examine their own.     

The golden age of biochemical research in photosynthesis

The perspectives and enthusiasms recorded in this review describe the events I witnessed and, in small ways, contributed to. Two great rewards emerged from my experiences – the pleasure of doing experiments and the great wealth of friendships with students and colleagues. As a graduate student, phenomena appeared at the bench before me which clarified the coupling of electron transport to ATP synthesis. My first PhD graduate student measured concentrations of pyridine nucleotides in chloroplasts and his results have been often confirmed and well used. All of the many graduate students who followed contributed to our understanding of photosynthesis. I have taken much pleasure from documenting the details of photosynthetic phosphorylation and electron transport in cyanobacteria. Studies of the `c' type cytochromes in these organisms continue to fascinate me. My experiences in government in its efforts to promote research are unusual, perhaps unique. A rare event outside the laboratory – a natural bloom of cyanobacteria – stimulated new thoughts and special opportunities for laboratory science. Photosynthesis seems magisterial in its shaping of our planet and its biology and in the details of its cleverness that were revealed in the time of my witness. This revised version was published online in June 2006 with corrections to the Cover Date.     

Being at the right place at the right time

I am tremendously honored to receive the 2012 Women in Cell Biology Junior Award. In this essay, I recount my career path over the past 15 years. Although many details are specific to my own experiences, I hope that some generalizations can be made to encourage more women to pursue independent scientific careers. Mine is a story of choosing a captivating question, making the most of your opportunities, and finding a balance with life outside the lab.It is a great honor to have been awarded the 2012 Women in Cell Biology Junior Award from the ASCB. Looking back at the 15 years I have spent doing research in cell biology, my overwhelming feeling is that it has been and still is a lot of fun. I am extremely fortunate to have a job that I truly enjoy and that gives me complete intellectual freedom. My lab choices over the years were motivated by scientific curiosity and enthusiasm for new environments and topics, rather than by career building. It is thus truly amazing to be rewarded for (rather a lot of) work enjoyed.     

Inclusion: an individual pursuit requiring organization-level investment to sustain it

If this was not happening in the midst of the COVID-19 pandemic, I imagine that I would be speaking these words instead of writing them on my laptop. Even so, I am so jazzed for this opportunity! No word or phrase describes what I am feeling in this moment in receiving the 2021 American Society for Cell Biology Prize for Excellence in Inclusivity. It is certainly an honor to be recognized in this way. I am grateful to the Howard Hughes Medical Institute for awarding me additional resources to keep on keeping on. My approach to finding the connection between people and their science certainly could use the monetary support. Resources open doors. At the same time that I am grateful for the attention, I am not exactly sure what to do with the spotlight. Importantly, there are a host of other folks out there also doing amazing things who have never been recognized. Let’s work to ensure that their contributions are supported, appreciated, and recognized. Instead of focusing the spotlight on me, I would rather redirect it to recognize my foundational influences. I also hope to encourage the need for institutional approaches beyond celebrating individual accomplishment.

O. A. Quintero‐CarmonaJo Rae Wright was my graduate advisor and the model for how I have tried to work with my students and colleagues to support their opportunities while also “doing science.” I wanted to start graduate school as soon as I could after graduating college, so after letting the Cell and Molecular Biology Program at Duke University know that I was accepting their offer, I started thumbing through their program booklet looking for labs with interesting research projects (a web presence wasn’t even really a thing for departments in 1996). I worked alphabetically and contacted a handful of labs one at a time to see whether anyone was willing to take on an early-rotation student. It was an unusual request for the way that the program had operated previously, and Jo Rae was the only person to agree to it. I don’t remember exactly, but she said something like, “We accepted you into the program, so I would be happy to host your first rotation.” The sense that I got was that, within the limits of her time and resources, she was willing to become my mentor because I needed one. She trusted the admissions process, so why not bring an eager student into the lab. I spent the summer settling in to the life of a graduate student—sort of.At first, I was bad at graduate school. I am curious about all sorts of things, which means I am also easily pulled in too many directions. In that first year of school I spent way too much time simply visiting other students in my cohort to see what it was that they were up to each day. I cannot imagine how distracting I must have been to them and probably extremely irritating to their PIs as well. If you were in Cell Biology at Duke in 1996–97, I am sincerely grateful that you tolerated my shenanigans. Where others might have taken me to task, Jo Rae looked for opportunities to redirect my energies more productively. She and another professor, Dan Kiehart, guided me toward participating in the Physiology Course at the Marine Biological Laboratory, where I learned what I needed to do to be a scientist in a way that would not have been possible otherwise. While there, I saw PIs working with students chasing the joy of discovery, and it felt like it was purely for the sake of a deeper understanding of biology and preparing the next generation of scientists to do the same. Resources gave us the liberty to focus on scientific discovery with minimal concern for where would be the highest profile place to publish. Although I acknowledge that the summer course environments may not be the most representative of the daily life of a scientist at a home institution, such an opportunity left a mark—I wanted to come as close as I could to emulating that environment when I got back to Duke and (eventually) when I had the chance to run a research group and teach students.Along the way, Jo Rae made sure to include me and my fellow lab mates in all aspects of the science. At national meetings she included us at every step, introducing us to her contemporaries and putting us in spaces where we would rub elbows with luminaries in the field. When we were in those environments, she made sure that I felt like a junior colleague. I cannot recall ever feeling like a “trainee.” Back home at Duke, I had opportunities to do everything that a scientist might do in addition to “sciencing.” Sure, I would write papers, contribute to grants, and be part of her review of papers. I was also encouraged to mentor undergraduates, teach, advocate for federal funding at the time of the National Institutes of Health (NIH) doubling, and plan events for Duke’s summer undergraduate research program, if I so chose. Similarly, when I expressed an interest in focusing on science with undergraduates, she was 100% on board with finding ways to combine my graduate school commitments with teaching and mentoring opportunities. Importantly, at a time when expressing interest in an “alternative career” was not always supported by faculty mentors, Jo Rae encouraged me to seek out only those potential postdoctoral mentors who would actively support my goals. Not only that, she went out of her way to find out what options I might have, which led to her learning about the NIH-funded Institutional Research and Academic Career Development Award postdoctoral programs in their first year of existence.In a sentence, because Jo Rae was 100% invested in including me in science by finding the framework that best suited my interests and potential, I grew into my success. This was a form of success that wasn’t decided by someone else; I had defined it for myself with Jo Rae serving as a true advisor in every sense of the word—she was in it for me. She helped to build the crucial foundations that helped me find the opportunities that matched my goals. As a result of her influence, I have also had the strength to make some critical, nontraditional choices along the way. Her mentorship style was tailored to each individual’s needs. She invested the time to figure out our strengths, and also learned which levers would motivate us to meet our potential. The members of her lab became successes because she helped all of us to both define success and achieve our own version of it. Such a personal approach is extremely powerful. Jo Rae passed away in 2012, and with her passing I lost the most important influence in my professional life. Duke University and the pulmonary physiology community lost an example for inclusive mentorship and a significant amount of capacity for such an approach. Since her passing, multiple awards have been established to honor Jo Rae’s legacy as an outstanding woman in science. I would argue that mentoring of junior colleagues may be a more significant legacy than her scientific output. Jo Rae is deserving of this award.Recognitions such as this one are an important way to amplify examples of what we often say we hope to achieve as a department, an institution, or a scientific society. However, if our focus is solely on the efforts of individuals, we are missing an opportunity. While I am humbled to be considered in the same league as the previous award recipients, we are each in our own way scrambling to do what we can while we can do it. When individuals have some positive outcomes, our institutions and organizations will celebrate what these folks have done as they have played some role in supporting these opportunities. Although what we do is worthwhile, it is really hard to do it successfully and sustainably without proper institutional support. We each face hinderances that can undermine the work that we want to take on. Burnout is a real outcome of doing the work that we care about and that our organizations publicly state is important. This is especially true in environments where that work is undervalued and underresourced. You do not have to do a very extensive internet search to identify where the institutions that have supported my work also have exclusionary legacies and current negative influences that continue to hinder their potential for broader, more meaningful progress. In many instances inclusion has yet to be baked into institutional culture in a way that impacts how organizations operate. Although I have had some institutional support to develop a career modeled on what I experienced under Jo Rae’s mentorship, the students and faculty at these institutions know that what gets headlines can often be an exceptional situation, rather than a typical everyday experience. Rather than showcasing the good work of individuals in their ranks, an organization should devote itself to furthering the idea that it is willing to make significant institutional investments in that good work. By building the internal infrastructure and capacity to support inclusion efforts, organizations would demonstrate that inclusion is an essential component of the institutional standard practice. The positive outcomes that this award is intended to highlight would then be a shared characteristic of the community. A shared vision paired with shared effort and resource-support might cut down on burnout of those currently carrying more than their share of the load.I imagine that the idea for these awards is to celebrate good work while also demonstrating to other individuals what is possible. With that in mind, if institutions worked at using the example of those in the vanguard as a way to build structures that value and support inclusive approaches, they would increase their own ability to serve their constituents. They may also influence other institutions to do the same. My graduate institution benefited from Jo Rae’s work while she was present and was beginning to institutionalize her view of inclusion in the last years of her life. As Dean of the Graduate School, the model for how she ran her lab informed her vision for graduate education campus-wide. She wanted to build a structure that would identify, recruit, and retain talent. She wanted to provide that talent with opportunities to become expert in how they wanted to contribute to the world. By ensuring that they had access to the relevant experiences and skills, she hoped to support them as they set themselves up for success as they defined it.I accept this award in honor of Jo Rae Wright, and on behalf of the students who have trusted me. All I have ever wanted was to be able to recreate for my undergraduates what Jo Rae had done for the people under her wing. I am building a career around that goal as part of a department keenly supportive of these efforts. My hope is that other individuals will develop their own approaches to inclusion because they find themselves in supportive institutional environments. More importantly, I would like to see organizations begin to truly prioritize inclusive approaches through funding and through policy. Institutions could make sufficient resources available to support inclusive efforts and allow creativity in how faculty mobilize those resources. Just as Jo Rae had the flexibility to adjust to our needs, institutional efforts will benefit when limited resource access is not a hindrance to inclusive excellence. Additionally, it will be critical to acknowledge the time and effort that such endeavors require in evaluating faculty contributions. It can no longer be the icing on the cake of a portfolio—developing inclusive capacity has to be recognized as an essential component of our work. Until these changes take root at the institutional level, this kind of work may shine brightly, but will continue to be stochastic and short-lived. All those efforts “will be lost in time, like tears in rain.” It is on all of us to prevent such a tragic ending.     

Finding a niche

Although I always knew I wanted to be a scientist, I didn't know I would become a cell biologist. Events in life that you would never have predicted can greatly impact your career trajectory. I have learned to let those events take me in new directions. Following a desire to investigate an understudied area of cell biology, I have found a niche. In this area, my lab is poised to contribute significantly toward understanding the fundamental molecular mechanisms underlying polarized plant cell growth.     

The Chemical Synthesis of DNA/RNA: Our Gift to Science

It is a great privilege to contribute to the Reflections essays. In my particular case, this essay has allowed me to weave some of my major scientific contributions into a tapestry held together by what I have learned from three colleagues (Robert Letsinger, Gobind Khorana, and George Rathmann) who molded my career at every important junction. To these individuals, I remain eternally grateful, as they always led by example and showed many of us how to break new ground in both science and biotechnology. Relative to my scientific career, I have focused primarily on two related areas. The first is methodologies we developed for chemically synthesizing DNA and RNA. Synthetic DNA and RNA continue to be an essential research tool for biologists, biochemists, and molecular biologists. The second is developing new approaches for solving important biological problems using synthetic DNA, RNA, and their analogs.     

The microenvironment matters

The physical and biochemical properties of the microenvironment regulate cell behavior and modulate tissue development and homeostasis. Likewise, the physical and interpersonal cues a trainee receives profoundly influence his or her scientific development, research perspective, and future success. My cell biology career has been greatly impacted by the flavor of the scientific environments I have trained within and the diverse research mentoring I have received. Interactions with physical and life scientists and trainees and exposure to a diverse assortment of interdisciplinary environments have and continue to shape my research vision, guide my experimental trajectory, and contribute to my scientific success and personal happiness.     

After the year of the dumpster fire

2020 has been one of the craziest and strangest years we have lived through. Now that it’s over, it’s an opportunity to show gratitude for all the good things. Subject Categories: S&S: History & Philosophy of Science

I moved to New York City the year of the attacks on September 11, 2001, one of the bleakest moments in the history of the United States. I was also in New York City when Superstorm Sandy hit in 2012. Luckily, much fewer people died due to the storm, but it was incredibly disruptive to many scientists in the affected area—my laboratory had to move four times over a period of 6 years in the storm’s aftermath. These were awful, tragic events, but 2020 may go down in the record books as one of the most stressful and crazy years in modern times. Not to be outdone, 2021 has started terribly as well with COVID‐19 still ravaging the world and an attack on the US Capitol, something I thought I’d never see in my lifetime. The unnecessary deaths and the damage to America’s “House of the People” were heartbreaking.While these events were surely awful, nothing will be as crushing as the deaths of family members, close friends, and the children of friends; perhaps, it is these experiences—and the death of a beloved dog—that prepared me for this year and made me grateful, maybe even more than usual, for what I have. But in the age of a pandemic, what am I particularly grateful for?I''m ridiculously grateful to have a job, a roof over my head, and food security. The older I get, the more I see illness and injury affect my colleagues, family, and friends, I increasingly appreciate my good health. I am grateful for Zoom (no, I have no investment in Zoom)—not for the innumerable seminars or meetings I have attended, but for the happy hours that helped to keep me sane during the lockdown. Some of these were with my laboratory, others with friends or colleagues, sometimes spread over nine time zones. Speaking of which, I’m also grateful for getting a more powerful router for the home office.I''m immeasurably grateful to be a scientist, as it allows me to satisfy my curiosity. While not a year‐round activity, it is immensely gratifying to be able to go to my laboratory, set up experiments, and watch the results coming in. Teaching and learning from students is an incredible privilege and educating the next generation of scientists how to set up a PCR or run a protein gel can, as a well‐known lifestyle guru might say, spark serious joy. For this reason, I’m eternally grateful to my trainees; their endless curiosity, persistence, and energy makes showing up to the laboratory a pleasure. My dear friend Randy Hampton recently told me he received a student evaluation, thanking him for telling his virtually taught class that the opportunity to educate and to be educated is something worth being grateful for, a sentiment I passed onto a group of students I taught this past fall. I believe they, too, were grateful.While all of the above things focus on my own life, there are much broader things. For one, I am so grateful to all of those around the globe who wear masks and keep their distance and thereby keep themselves and others safe. I am grateful for the election of an American president who proudly wears a mask—often quite stylishly with his trademark Ray‐Ban Aviators—and has made fighting the COVID‐19 pandemic his top priority. President Biden''s decision to ramp up vaccine production and distribution, along with his federal mask mandate, will save lives, hopefully not just in the United States but worldwide.This Gen‐X‐er is also especially grateful to the citizens of Generations Y and Z around the world for fighting for social justice; I am hopeful that the Black Lives Matter movement has got traction and that we may finally see real change in how communities of color are treated. I have been heartened to see that in my adopted home state of New York, our local politicians ensure that communities that have been historically underserved are prioritized for COVID‐19 testing and vaccinations. Along these lines, I am also incredibly encouraged by the election of the first woman who also happens to be of African and Asian heritage to the office of vice president. Times are a changin''...While it is difficult to choose one, top thing to be grateful for, I would personally go for science. I am stoked that, faced with a global crisis, science came to the rescue, as it often has in the past. If I had to find a silver lining in COVID‐19—albeit it would be for the darkest of clouds—I am grateful for all of our colleagues, who despite their usual arguing, quickly and effectively developed tests, provided advice, epidemiological data and a better understanding of the virus and its mode of infection, and ultimately developed therapies and vaccines to save lives. The same can be said for the biotech and pharmaceutical industry that, notwithstanding its often‐noted faults, has been instrumental in developing, testing and mass‐producing efficient and safe vaccines in blistering, record time. Needless to say, I have also much gratitude to all of the scientists and regulators at the FDA and elsewhere who work hard to make life as we once knew it come back to us, hopefully in the near future.Once again, thank you for everything, Science.     

A Love Affair with Bacillus subtilis

My career in science was launched when I was an undergraduate at Princeton University and reinforced by graduate training at the Massachusetts Institute of Technology. However, it was only after I moved to Harvard University as a junior fellow that my affections were captured by a seemingly mundane soil bacterium. What Bacillus subtilis offered was endless fascinating biological problems (alternative sigma factors, sporulation, swarming, biofilm formation, stochastic cell fate switching) embedded in a uniquely powerful genetic system. Along the way, my career in science became inseparably interwoven with teaching and mentoring, which proved to be as rewarding as the thrill of discovery.     

Plans of mice and men: from bench science to science policy

The transition from bench science to science policy is not always a smooth one, and my journey stretched as far as the unemployment line to the hallowed halls of the U.S. Capitol. While earning my doctorate in microbiology, I found myself more interested in my political activities than my experiments. Thus, my science policy career aspirations were born from merging my love of science with my interest in policy and politics. After receiving my doctorate, I accepted the Henry Luce Scholarship, which allowed me to live in South Korea for 1 year and delve into the field of science policy research. This introduction into science policy occurred at the South Korean think tank called the Science and Technology Policy Institute (STEPI). During that year, I used textbooks, colleagues, and hands-on research projects as my educational introduction into the social science of science and technology decision-making. However, upon returning to the United States during one of the worst job markets in nearly 80 years, securing a position in science policy proved to be very difficult, and I was unemployed for five months. Ultimately, it took more than a year from the end of the Luce Scholarship to obtain my next science policy position with the American Society for Microbiology Congressional Fellowship. This fellowship gave me the opportunity to work as the science and public health advisor to U.S. Senator Harry Reid. While there were significant challenges during my transition from the laboratory to science policy, those challenges made me tougher, more appreciative, and more prepared to move from working at the bench to working in the field of science policy.     

A passion for the science of the human genome

The complete sequencing of the human genome introduced a new knowledge base for decoding information structured in DNA sequence variation. My research is predicated on the supposition that the genome is the most sophisticated knowledge system known, as evidenced by the exquisite information it encodes on biochemical pathways and molecular processes underlying the biology of health and disease. Also, as a living legacy of human origins, migrations, adaptations, and identity, the genome communicates through the complexity of sequence variation expressed in population diversity. As a biomedical research scientist and academician, a question I am often asked is: “How is it that a black woman like you went to the University of Michigan for a PhD in Human Genetics?” As the ASCB 2012 E. E. Just Lecturer, I am honored and privileged to respond to this question in this essay on the science of the human genome and my career perspectives.

“Knowledge is power, but wisdom is supreme.”

     

An unconventional route to becoming a cell biologist

I am honored to be the E. B. Wilson Award recipient for 2015. As we know, it was E. B. Wilson who popularized the concept of a “stem cell” in his book The Cell in Development and Inheritance (1896, London: Macmillan & Co.). Given that stem cell research is my field and that E. B. Wilson is so revered within the cell biology community, I am a bit humbled by how long it took me to truly grasp his vision and imaginative thinking. I appreciate it deeply now, and on this meaningful occasion, I will sketch my rather circuitous road to cell biology.I grew up in a suburb of Chicago. My father was a geochemist, and for everyone whose parents worked at Argonne National Laboratories, Downers Grove was the place to live. My father’s sister was a radiobiologist and my uncle was a nuclear chemist, both at Argonne; they lived in the house next door. Across the street from their house was the Schmidtke’s Popcorn Farm—a great door to knock on at Halloween. The cornfields were also super for playing hide-and-seek, particularly when you happened to be shorter than those Illinois cornstalks.Open in a separate windowElaine FuchsI remember when the first road in the area was paved. It made biking and roller-skating an absolute delight. Fields of butterflies were everywhere, and with development came swamps and ponds filled with pollywogs and local creeks with crayfish. It was natural to become a biologist. When coupled with a family of scientists and a mother active in the Girl Scouts, all the resources were there to make it a perfect path to becoming a scientist.I could hardly wait until I was in junior high school, when I could enter science fairs. You would think that my science-minded family might help me choose and develop a research project. True to their mentoring ethos, they left these decisions to me. My first project was on crayfish behavior. I recorded the response of the crayfish I had caught to “various external stimuli.” At the end of this assault, I dissected the crayfish and, using “comparative anatomy,” attempted to identify all the parts. The second project was no gentler. I focused on tadpole metamorphosis and the effects of thyroid hormone in accelerating development at low concentrations and death at elevated concentrations. Somehow, I ended up going all the way to the state fair, where it became clear that I had serious competition. That experience, however, whetted my appetite to gain more lab experience and to learn to read the literature more carefully.My experience with high school biology prompted me to gravitate toward chemistry, physics, and math. When it came to college, my father told me that if there was a $2000/year (translated in 2015 to be $30,000/year) reason why I should go anywhere besides the University of Chicago (where Argonne scientists received a 50% tuition cut for their children) or the University of Illinois (then $200/year tuition), we could “discuss” it further. Having a sister, father, aunt, and uncle who went to the University of Chicago, I chose the University of Illinois and saved my Dad a bundle of money. At Illinois, I thought I might revisit biology, but my choices for a major were “biology for teachers” or “honors biology.” The first did not interest me; the second seemed intimidating.I enrolled as a chemistry major. Four years went by, during which time I never took a biology class. I enjoyed quantum mechanics, physics, and differential equations, and problem solving became one of my strengths. In the midst of the Vietnam War era, however, Illinois was a hotbed of activity. I was inspired to apply to the Peace Corps, with a backup plan to pursue science that would be more biomedically relevant than quantum mechanics. I was accepted to go to Uganda with the Peace Corps, but with Idi Amin in office, my path to science was clear. Fortunately, the schools I applied to accepted me, even though, in lieu of GRE scores, I had submitted a three-page essay on why I did not think another exam was going to prove anything. I chose Princeton’s biochemistry program. This turned out to be a great, if naïve choice, as only after accepting their offer did I take a biochemistry class to find out what I was getting into. I chose to carry out my PhD with a terrific teacher of intermediary metabolism, Charles Gilvarg, who worked on bacterial cell walls. My thesis project was to tackle how spores break down one cell wall and build another as they transition from quiescence to vegetative growth.By my fourth year of graduate school, I was trained as a chemist and biochemist and was becoming increasingly hooked on biomedical science. I listened to a seminar given by Howard Green, who had developed a method to culture cells from healthy human skin under conditions in which they could be maintained and propagated for hundreds of generations without losing their ability to make tissue. At the time, Howard referred to them as epidermal keratinocytes, but in retrospect, these were the first stem cells ever to be successfully cultured. I was profoundly taken by the system, and Howard’s strength in cell biology inspired me. It was the perfect match for pursuing my postdoctoral research. The time happened to be at the cusp of DNA recombinant technology.At MIT, I learned how to culture these cells. I wanted to determine their program of gene expression and how this changed when epidermal progenitors embark on their terminal differentiation program. While the problem in essence was not so different from that of my graduate work at Princeton, I had miraculously managed to receive my PhD without ever having isolated protein, RNA, or DNA. Working in a quintessential cell biology lab and tackling a molecular biology question necessitated venturing outside the confines of the Green lab and beyond the boundaries of my expertise. Fortunately, this was easy at MIT. Richard Hynes, Bob Horvitz, Bob Weinberg, and Graham Walker were all assistant professors, and their labs were very helpful, as were those of David Baltimore and Phil Sharp, a mere walk across the street. On the floor of my building, Steve Farmer, Avri Ben Ze’ev, Gideon Dreyfuss, and Ihor Lemischka were in Sheldon Penman’s lab just down the hall, and they were equally interested in mRNA biology, providing daily fuel for discussions. Uttam Rhajbandary’s and Gobind Khorana’s labs were also on the same floor, making it easy to learn how to make oligo(dT)-Sepharose to purify my mRNAs. Vernon Ingram’s lab was also on the same floor, so learning to make rabbit reticulocyte lysates to translate my mRNAs was also possible. Howard bought a cryostat, so I could section human skin and separate the layers. And as he was already working with clinicians at Harvard to apply his ability to create sheets of epidermal cells for the treatment of burn patients, I had access to the leftover scraps of human tissue that were also being used in these operations.The three years of my postdoc were accompanied by three Fuchs and Green papers. The first showed that epidermal keratinocytes spend most of their time expressing a group of keratin proteins with distinct sequences. The second showed that these keratins were each encoded by distinct mRNAs. The third showed that, as epidermal keratinocytes commit to terminally differentiate, they switch off expression of basal keratins (K5 and K14) and switch on the expression of suprabasal keratins (K1 and K10). That paper also revealed that different stratified tissues express the same basal keratins but distinct sets of suprabasal keratins. I am still very proud of these accomplishments, and my MIT experience made me thirst to discover more about the epidermis and its stem cells.My first and only real job interview came during my second year of postdoc, at a time when I was not looking for a job. I viewed the opportunity, initiated by my graduate advisor, as a free trip home to visit my parents and my trial run to prepare me for future searching. I was thrilled when this interview materialized into an offer to join the faculty, for which the University of Chicago extended my start time to allow me to complete my three years with Howard.Times have clearly changed, and it is painful to see talented young scientists struggle so much more today. That said, I have never looked ahead very far, and having a lack of expectations or worry is likely to be as helpful today as it was then. I am sure it is easier said than done, but this has also been the same for my science. I have always enjoyed the experiments and the joy of discovery. There was no means to an end other than to contemplate what the data meant in a broader scope.I arrived at the University of Chicago with a well-charted route. My aim was to make a cDNA library and clone and characterize the sequences and genes for the differentially expressed keratins I had identified when I was at MIT. It was three months into my being at Chicago when my chair lined up some interviews for me to hire a technician. I was so immersed in my science that I did not want to take time to hire anyone. I hired the first technician I interviewed. Fortunately, it worked out. However, I turned graduate students away the first year, preferring to carry out the experiments with my technician and get results. After publishing two more papers—one on the existence of two types of keratins that were differentially expressed as pairs and the other on signals that impacted the differential expression of these keratin pairs, I decided to accept a student, who analyzed the human keratin genes. My first postdoc was a fellow grad student with me at Princeton; she studied signaling and keratin gene expression. My second postdoc was initiated by my father, who chatted with him at the elevator when I was moving into my apartment. He set up DNA sequencing and secondary-structure prediction methods, and the lab stayed small, focused, and productive.I was fascinated by keratins, how they assembled into a network of intermediate filaments (Ifs). When thalassemias and sickle cell anemia turned out to be due to defects in globin genes, I began to wonder whether there might be human skin disorders with defective keratin genes. I had no formal training in genetics, and there were no hints of what diseases to focus on. Thus, rather than using positional cloning to identify a gene mutation associated with a particular disease, we took a reverse approach: we first identified the key residues for keratin filament assembly. After discovering that mutations at these sites acted dominant negatively, we engineered transgenic mice harboring our mutant keratin genes and then diagnosed the mouse pathology. Our diagnoses, first for our K14 mutations and then for our K10 mutations, turned out to be correct: on sequencing the keratins from humans with epidermolysis bullosa simplex (EBS), we found K14 or K5 mutations; similarly, we found K1 or K10 mutations in affected, but not in unaffected, members of families with epidermolytic hyperkeratosis (EH). Both are autosomal-dominant disorders in which patients have skin blistering or degeneration upon mechanical stress. Without a proper keratin network, the basal (EBS) or suprabasal (EH) cells could not withstand pressure. Ironically, family sizes of all but the mildest forms of these disorders were small, meaning that the disorders were not amenable to positional cloning. But the beauty of this approach is that once we had made the connection to the diseases, we understood their underlying biology. In addition, the IF genes are a superfamily of more than 100 genes differentially expressed in nearly all tissues of the body. Once we had established EBS as the first IF gene disorder, the pathology and biology set a paradigm for a number of diseases of other tissues that turned out to be due to defects in other IF genes.Fortunately, I had students, Bob Vassar (professor, Northwestern University) and Tony Letai (associate professor, Harvard Medical School), and a postdoc, Pierre Coulombe (chair, Biochemistry and Molecular Biology, Johns Hopkins University), who jumped into this fearless venture with me. We had to go off campus to learn transgenic technology. I had never worked with mice before. When Bob returned to campus with transgenic expertise, we hired and trained Linda Degenstein, whose love for animal science was unparalleled. Pierre’s prior training in electron microscopy was instrumental in multiple ways. Additionally, I was not a dermatologist and had no access to human patients. Fortunately Amy Paller, MD, at Northwestern volunteered to work with us.The success of this project attests to an important recipe: 1) Pursue a question you are passionate about. 2) In carrying out rigorous, well-controlled experiments, each new finding should build upon the previous ones. 3) If you have learned to be comfortable with being uncomfortable, then you will not be afraid to chart new territory when the questions you are excited to answer take an unanticipated turn. 4) Science does not operate in a vacuum. Interact well with your lab mates and take an interest in their science as well as your own. And wherever you embark upon a pathway in which the lab’s expertise is limited, do not hesitate to reach out broadly to other labs and universities.I have followed this recipe now for more than three decades, and it seems to work pretty well. A lab works only when its students and postdocs are interactive, naturally inquisitive, and freely share their ideas and findings. I have been blessed to have a number of such individuals in my lab over the years. When push comes to shove, I am always inclined first to shave from the “brilliant” category and settle for smart, nice people who are passionate and interactive about science and original and unconventional in their thinking.So what questions have I been most passionate about? I have always been fascinated with how tissues form during development, how they are maintained in the adult, and how tissue biology goes awry in human disorders, particularly cancers. I first began to think about this problem during my days at Princeton. I also developed a dogma back then that I still hold: to understand malignancies, one must understand what is normal before one can appreciate what is abnormal. I think this is why I have spent so much of my life focusing on normal tissue morphogenesis, despite my passion for being at the interface with medicine. And because skin has so many amazingly interesting complexities, and because it is a great system to transition seamlessly between a culture dish and an animal, I have never found a reason to choose any other tissue over the one I chose many years back.I will not dwell on the various facets of skin biology we have tackled over the years. Our initial work on keratins was to obtain markers for progenitors and their differentiating lineages. This interest broadened to understanding how proliferative progenitors form cytoskeletal networks and how the cytoskeleton makes dynamic rearrangements during tissue morphogenesis.From the beginning, the lab has also been fascinated by how tissue remodeling occurs in response to environmental signals. Indeed, signals from the microenvironment trigger changes in chromatin dynamics and gene expression within tissue stem cells. Ultimately, this leads to changes in proteins and factors that impact on cell polarity, spindle orientation, asymmetric versus symmetric fate specifications, and ultimately, the balance between proliferation and differentiation.The overarching theme of my lab over these decades is clear, namely, to understand the signals that unspecified progenitors receive that instruct them to generate a stratified epidermis, make hair follicles, or make sweat and sebaceous glands. And if we can understand how this happens, then how are stem cells born, and how do they replace dying cells or regenerate tissue after injury? And, finally, how does this process change during malignant progression or in other aberrant skin conditions?In tackling tissue morphogenesis, I have had to forgo knowing the details of each tree and instead focus on the forest. There are many times when I stand back and can only admire those who are able to dissect beautiful cellular mechanisms with remarkable precision. But I crossed that bridge some years ago in tackling a problem that mandates an appreciation of nearly all the topics covered in Bruce Alberts’ textbook Molecular Biology of the Cell. I am now settled comfortably with the uncomfortable, and the problem of tissue morphogenesis in normal biology and disease continues to keep me more excited about each year’s research than I was the previous year. Perhaps the difference between my days as a student, postdoc, and assistant professor and now is that my joy and excitement is as strong for those I mentor and have mentored as it is for myself.     

Biophysical Reviews’ “Meet the Councilor Series”—a profile of Peter Pohl

It is my pleasure to write a few words to introduce myself to the readers of Biophysical Reviews as part of the “Meet the Councilor Series.” Currently, I am serving the second period as IUPAB councilor after having been elected first in 2017. Initially, I studied Biophysics in Moscow (Russia) and later Medicine in Halle (Germany). My scientific carrier took me from the Medical School of the Martin Luther University of Halle-Wittenberg, via the Leibniz Institute for Molecular Pharmacology (Berlin) and the Institute for Biology at the Humboldt University (Berlin) to the Physics Department of the Johannes Kepler University in Linz (Austria). My key research interests lie in the molecular mechanisms of transport phenomena occurring at the lipid membrane, including (i) spontaneous and facilitated transport of water and other small molecules across membranes in reconstituted systems, (ii) proton migration along the membrane surface, (iii) protein translocation, and (iv) bilayer mechanics. Training of undergraduate, graduate, and postdoctoral researchers from diverse academic disciplines has been—and shall remain—a consistent part of my work.

     

Gaia Pigino: Inside the cell

Gaia Pigino studies the molecular mechanisms and principles of self-organization in cilia using 3D cryo-EM.

Gaia Pigino was only 3 yr old when she became fascinated with nature in the beautiful countryside of Siena, Italy, where she grew up. The neighbor’s daughter showed her a hen in the chicken coop, and they caught it in the act of laying an egg. Gaia remembers, “This was for me almost a shock, as my experience about eggs was that they come directly out of paper boxes!” Her father was also an important part of awakening Gaia’s curiosity for the amazing things in nature. He used to bring home the award-winning magazine Airone, the Italian equivalent of National Geographic. Gaia never missed an issue; even before learning to read, she could spend hours looking at the captivating photos of the wildlife. She wanted to understand what she was seeing, and maybe because of that, she was determined to do science.Gaia Pigino. Photo courtesy of Human Technopole.Gaia took her first “scientific” steps with Professor Fabio Bernini and Professor Claudio Leonzio at the University of Siena, where she studied bioindicators of soil contamination and detoxification strategies of soil arthropods as part of her PhD project. But it was later, when she joined the laboratory of Professor Pietro Lupetti and met Professor Joel Rosenbaum, a pioneer of cilia research, that Gaia discovered the world of 3D EM and felt her place was “inside a single cell.” She solidified her interest in the structure of protein complexes of cilia and flagella and boosted her passion for cryo-electron tomography (ET) in the laboratory of Professor Takashi Ishikawa, first at the ETH Zurich and then at the Paul Scherrer Institut in Switzerland. In 2012, Gaia started her own laboratory at the Max Planck Institute of Molecular Cell Biology and Genetics in Dresden, Germany, with the vision of creating a truly interdisciplinary laboratory. Her team combines techniques from different fields such as biophysics, cell biology, and structural biology to answer open questions in the cilia field. Gaia recently moved countries again—this time to take over the position of Associate Head of the Structural Biology Research Centre, at the Human Technopole, Milan, Italy.We reached out to Gaia to learn more about her scientific journey and future research directions.What interested you about cilia?The first thing that attracted me toward cilia and flagella were some EM micrographs, by Professor Romano Dallai in Siena, that showed the beautiful geometrical microtubular structures of sperm flagella. I was intrigued by the apparent perfection of these organelles that clearly showed me that a cell is a coordinated system of complex molecular machines, the mechanism of many of which we do not understand. Soon after, Professor Joel Rosenbaum introduced me to the bidirectional transport of components inside cilia, which, he explained to me, is required for both assembly and function of virtually all cilia and flagella, from the motile cilia in our lungs to the primary cilium in our kidneys. He called it intraflagellar transport (IFT) and compared it to a Paternoster elevator, where the individual cabins were what we now call IFT trains. I was completely fascinated by the IFT system, the structure, the function, the dynamics, and the mechanism of which were still largely unknown. Quickly, I realized that in addition to IFT, cilia represent a virtually infinite source of open biological questions waiting to be solved, from the mechanics and regulation of the beating to the sensory function of primary cilia, and their importance for human health.What are some of the scientific questions currently of interest in your laboratory?In the past few years, we have made substantial contributions to the current understanding of the structure and the mechanism of the IFT (1, 2, 3). Currently, we are investigating how the structure of IFT trains relates to their functions by looking, in cryo-electron tomography, at how anterograde trains transform into retrograde trains and at how different ciliary cargoes are loaded on the trains. Beside this more classical line of research, we are exploring other approaches to study IFT, for instance we have developed a method to reactivate IFT trains in vitro on reconstituted microtubules. We want to use this approach to investigate the behavior of IFT trains, and their motors, in experimentally controllable conditions, e.g., in the presence of only certain tubulin posttranslational modifications. We have also made interesting discoveries about the distribution of tubulin posttranslational modifications on the microtubule doublets of the axoneme and how this spatially defined tubulin code affects the function of different ciliary components. We hope we will be able to share these new “stories” with the structural and cell biology community very soon!What kind of approach do you bring to your work?I believe that the main reason for why science became an integral, and dominant, part of my life is because it provides infinite riddles and continuous challenges. I have always been curious about how things work in nature, but I quickly realized that learning from books didn’t satisfy me. My desire was to be at the frontline, to be among the ones that see things happening in front of their eyes, at the microscope, for the first time. I wanted to be among the ones that make the discoveries that students read about in textbooks. Thus, what I bring to my work is an endless desire of solving biological riddles, curiosity, creativity, determination, and energy, with which I hope to inspire the members of my team. My laboratory uses an interdisciplinary approach; we use whatever method, technique or technology is needed to reach our goal, from the most basic tool to the most sophisticated cryo-electron microscope. And if the method we need does not yet exist, we try to invent it.A young Gaia Pigino (3 yr old) the day she discovered how eggs are made. Photo courtesy of Giancarlo Pigino.Could you tell us a bit about the Structural Biology Research Centre at the Human Technopole (HT)?At the HT Structural Biology Centre, we are working to create a vibrant and interdisciplinary scientific environment that will attract molecular, structural, cell, and computational biologists from all over the world. We are creating fantastic facilities, including one of the most well equipped and advanced electron microscopy facilities in Europe—and likely the world—headed by Paolo Swuec. My team, together with the teams of my colleague Alessandro Vannini and the research group leaders Ana Casañal, Francesca Coscia, and Philipp Erdmann, already cover a vast range of competences and know-how from classical molecular and structural biology approaches, such as crystallography and protein biophysics, to cryo-CLEM, cryo-FIB SEM and cryo-ET, all of which allow us to address questions in cell biology. Our goal is to create a scientific infrastructure and culture that will enable biologists to obtain a continuum of structural and functional information across scales.What did you learn during your PhD and postdoc that helped prepare you for being a group leader? What were you unprepared for?I learned that everyday research is mostly made of failures, but that with the right amount of obsession, persistence, curiosity, and creativity, it is always possible to succeed and discover new things. Being given the freedom to develop your own ideas and your own project very early in your career is a treat; science is not only about having good ideas! One needs to follow up on these ideas with intense work and troubleshooting to make them reality. In addition, I realized that being fearless and attempting what is considered too difficult by others, despite challenges, can turn into a worthy learning experience. Also, how you present your work to the scientific community matters for swinging the odds of success in your favor. Different places might work in very different ways, and conducting good science does not only depend on you, but also on the possibilities given to you by your environment.What was I unprepared for?—I guess several things, but one comes immediately to mind: I underestimated how much being responsible not only for my own life and career, but also the career of students, postdocs, and others in the laboratory, would affect me personally.Structure of the 96-nm axonemal repeat reconstructed by cryo-ET and subtomogram averaging. Image courtesy of Gonzalo Alvarez Viar, Pigino Lab.What has been the biggest accomplishment in your career so far?This is a tricky question for me... I tend to look into the future more than celebrating the past. I fight to succeed in something, but as soon as I conquer it, I find it less of an achievement than the thing I could conquer next. Nevertheless, I am happy about the discoveries and the papers published together with my students and postdocs (1, 2, 3, 4, 5). I am extremely excited about the fact that after many years of work I am now leading an interdisciplinary laboratory, where we combine techniques from different fields. I am also happy that three times my husband and I were able to move from one world class academic institution to the another to start exciting and fitting jobs and could still live together in the same place. We worked hard for this, but we also got lucky.What has been the biggest challenge in your career so far?I studied French in school; I had almost no exposure to spoken English until the end of my PhD. To avoid having to show my English insufficiencies, I did hide beside the board of my poster at the first international conference I attended in 2004! It took me a while to overcome this barrier and feel confident to express my thoughts and ideas in English.What do you think you would be if you were not a scientist?I had been a good fencer during my youth. I was a member of the Italian National Team between ages 14 and 19 and saw quite a bit of the world, which was cool! When my sporting career failed, due to diabetes, I was torn between art and science. I guess that in a parallel universe, I am a wildlife photographer and a potter specialized in wood kiln firing. [Gaia confesses that she misses “the amazing and addictive adrenaline rush of a good fencing match!”]Any tips for a successful research career?Do not compare your performances to the ones of the people at your career stage; compare yourself with people that are already successful one level higher than you currently are at. For example, if you are a PhD student, ask yourself what in your current performance separates you from being a good postdoc—once a postdoc, what is missing to be a good PI.     

Building a science in Japan: The formative decades of Molecular Biology

I am honored to have been invited to participate in this Workshop on Comparative Studies of Building Molecular Biology, with a discussion of Japanese experiences in constructing a science — in this case, the discipline of molecular biology. As I understand it, the construction of a science must be equivalent to building a new culture. My having given this title to my paper suggests that I have enough knowledge about the subject to perhaps even extrapolate its course into the future — which I do not. What I do have is a sincere admiration of my old friends and colleagues, in Japan and elsewhere, who together tried to build a new science of molecular biology in Japan.     

Contributions of an animal scientist to reproductive biology

I became interested in biology as an undergraduate in a premedical curriculum but developed a passion for the field of reproductive biology because of a course in physiology of reproduction taken to meet requirements for admission to veterinary school. My career path changed, and I entered graduate school, obtained the Ph.D., and have enjoyed an academic career as a reproductive biologist conducting research in uterine biology and pregnancy in animal science departments at the University of Florida and at Texas A&M University. However, I have never allowed academic boundaries to interfere with research and graduate education as that is contrary to collegiality, the cornerstone of great universities. I consider that my major contributions to science include 1) identification of proteins secreted by cells of the uterine endometrium that are critical to successful establishment and maintenance of pregnancy; 2) discovery of steroids and proteins required for pregnancy recognition signaling and their mechanisms of action in pigs and ruminant species; 3) investigation of fetal-placental development and placental transport of nutrients, including water and electrolytes; 4) identification of linkages between nutrition and fetal-placental development; 5) defining aspects of the endocrinology of pregnancy; and 6) contributing to efforts to exploit the therapeutic value of interferon tau, particularly for treatment of autoimmune diseases. My current studies are focused on the role of select nutrients in the uterine lumen, specifically amino acids and glucose, that affect development and survival of the conceptus and translation of mRNAs and, with colleagues at Seoul National University, gene expression by the avian reproductive tract at key periods postovulation. Another goal is to understand stromal-epithelial cell signaling, whereby progesterone and estrogen act via uterine stromal cells that express receptors for sex steroids to stimulate secretion of growth factors (e.g., fibroblast growth factors and hepatocyte growth factor) that, in turn, regulate functions of uterine epithelial cells and conceptus trophectoderm.