®ESHRE. David Adamson Photography

Beatriz Rodríguez Alonso is a PhD student at University College Dublin. Her research group studies the interaction between early embryo and the oviduct in the bovine model.

Embryologist Media has contacted her to tell us about her career and her research on embryo-maternal communication.

“Embryology fulfilled all my requirements, and I would say it even surpassed all my expectations.”

Embryologist Media: Welcome Beatriz. First, we would like to know you a little more. Could you tell us a bit about your first years of training as a biologist?

Beatriz Rodríguez: Thank you very much! First of all, I would like to thank you for the opportunity to attend this interview with you. I have been following you since the very beginning and it seems to me that you have a great project on your hands.

I remember my first years of the Biology Degree at Universidad Autónoma de Madrid with great enthusiasm. I loved what I studied, although in the beginning I did not know what kind of job I wanted to have in the future… What I did know was that I enjoyed working in the laboratory, as well as the health branch.

EM: What was your motivation to lean towards the study of assisted reproduction?

BR: During the last stage of my undergraduate studies, I had the opportunity to attend a master class by a professional in the field of human reproduction as part of the “Embryology” module. Until that moment I did not even know biologists could end up performing such tasks. Then it was clear to me: Embryology fulfilled all my requirements, and I would say it even surpassed all my expectations!

EM: Also during this stage you did some research on sperm DNA fragmentation and oxidative stress. What did these studies entail?

BR: Thanks to the discovery of the professional career of the biologist in the world of embryology and assisted reproduction, I began to move around and look for any opportunity in this field. I managed to focus on the profession even before finishing my degree; I spoke with a reproduction clinic in Madrid (Clínica Tambre) and a professor at university and I performed my Final Degree Project on sperm DNA fragmentation. The time invested eventually paid off: two scientific papers were published in national journals, which included results from my project. In fact, one of them was awarded the Best Original Paper by ASESA, the Spanish Andrology Association.

Basically, we tried to analyse the relationship between oxidative stress and sperm DNA fragmentation. Initially, we studied two clinical features (varicocele and leukocytospermia) related to high levels of oxidative stress. We observed a relationship between these features and the increase in the levels of basal DNA fragmentation. In a second study, we measured the levels of oxidative stress in seminal plasma (by using the NBT-t test) and related those to the dynamics of sperm DNA fragmentation. Unfortunately, DNA fragmentation is not static; rather, it increases with the time of exposure to the source of oxidative stress.

EM: You have worked in several laboratories of assisted human reproduction. Could you tell us about your experience as an embryologist so far?

BR: My first contact took place during the internship of my Master Degree in Human Reproduction, taught at Universidad Complutense de Madrid. This internship was carried out at the Reproduction Unit of Sanitas (Madrid), where I had the opportunity to stay for longer. This allowed me to improve my skills in both the andrology and embryology laboratories. Later, I was hired at Clínica Tambre (Madrid), where I spent two years working in the andrology laboratory.

“I think one of the things I like most about my job is that there is not a typical day, because it depends on where you are.”

EM: Later on, you started your doctoral studies thanks to a Marie Curie Fellowship. What led you to start your doctorate? Was there any relevant experience that guided you towards the research world?

BR: I think there were several factors that motivated me to make the decision. Firstly, I am lucky to dedicate myself to something I am passionate about, and doing the PhD was an opportunity to go deeper into the field. Secondly, I started working in the andrology laboratory at Clínica Tambre. Finally, I was aware that doing a PhD may open doors for me in the future, so I could probably be back in research or teaching at university, or even access to relevant positions in assisted reproduction laboratories.

EM: Would you mind sharing with us your current research project theme?

BR: My research aims to improve our knowledge of embryo-maternal communication during the preimplantation stages of pregnancy. Specifically, I study the communication between the early embryo and the oviduct tube using the bovine model. My long-term objective is to understand the hormonal, cellular and molecular mechanisms regulating embryo growth and survival and interaction with the maternal reproductive tract in order to ensure successful establishment and maintenance of pregnancy.

“This type of work involves many complications at a technical level, and many people have been actively involved so that it could move forward.”

EM: How would you describe a typical day of your job as a researcher?

BR: I think one of the things I like most about my job is that there is not a typical day, because it depends on where you are. I have performed in vivo experiments in the bovine model in the magnificent facilities of University College Dublin: Lyons Research Farm. Then, in vitro experiments were carried out in the laboratory at the INIA (Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria in Madrid, Spain) using both embryo and oviductal cell cultures. Finally, when you are not performing or collaborating in any experiment, there goes “computer work”; you need to spend a lot of time reading, studying, writing… in other words, doing your thesis!

EM: What piece of advice would you give to someone who could be considering research in the field of assisted reproduction?

BR: It is a very personal decision. As for myself, it has been a full hit. I wanted to make a parenthesis in my career as a clinical embryologist to obtain my PhD, and then I may pick it up again eventually. However, I love the research world so much that my intention is to try to make my mark on it. In any case, and regardless of whether I succeed, I already consider having started my PhD a success. I believe that after what I have learned within the last years, I would probably face my work as a clinical embryologist from a better position, both in terms of knowledge and opportunities.

“Even around 20% of patients who attend assisted reproduction treatments are diagnosed with “sterility of unknown origin”. This means there is still much to improve on diagnosis, and the same goes for treatments.”

EM: You have recently presented your data on the transcriptomic response of bovine oviduct epithelial cells to embryonic contact. How did this study arise?

BR: It is actually within the research line of our group, which focuses on embryo-maternal communication during pre-implantation development. Specifically, we analyse the interaction that takes place between the embryo and the oviduct (Fallopian tube), where the embryo goes through the first 3-4 days of development until it reaches the uterus. In this study, our goal was to find out whether the embryo can regulate gene expression in oviduct cells. In other words, we wanted to know the effect the embryo may exert on the oviduct.

EM: In your opinion, what was the greatest technical difficulty in planning this study?

BR: To date, most of the studies that have described the effect of the embryo on the oviduct have been performed in multiovulatory species in which the embryo’s signal on the oviduct is amplified by the presence of several embryos. Regarding monovulatory species, so far it has been only possible to observe these effects in mares. My group works with the bovine model (cows), and the only way we have had so far to study this effect was to transfer multiple embryos into the oviduct. This approach has given us a lot of information, although these were obviously not physiological conditions. Our working hypothesis was that a single embryo may have the capacity to elicit a local response in the oviduct. The biggest difficulty we have encountered has been working with small sections of oviduct of about 2 cm, which is roughly 20-30 cm in cows, trying to find the embryo and working with a smaller proportion of oviductal cells to detect potential effects at a local level. Fortunately, results have been positive and we have finally been able to detect the effect of a single embryo on the cow oviduct.

EM: Would it be possible to study such interactions in vitro?

BR: Nowadays everything is about improving the in vitro models to obtain information about different processes, thus avoiding the use of live animals. In this study, we performed parallel experiments with an in vitro model by culturing oviductal cells to form a monolayer and co-culturing them with embryos. Unfortunately, we have not found similar results in vivo and in vitro. This suggests we must keep working and improving in vitro models, since there is a wide range of variation in the efficiency of each one of them. On the other hand, in vivo experiments currently provide unique and very relevant information to understand what happens under physiological conditions and in the real context of those cells within the body.

“We analyse the interaction that takes place between the embryo and the oviduct (Fallopian tube), where the embryo goes through the first 3-4 days of development until it reaches the uterus”.

EM: I suppose that in order to carry out these experiments on research animals, you must be given consent by an ethical committee that regulates these practices. Have you encountered any obstacle on this issue?

BR: Any person who is to make use of animals for an experiment must count with the appropriate consent granted by different regulatory organizations. I carried out these experiments in Ireland, where before performing an in vivo experiment you must obtain the Professional Certificate in Training on Experimental Animal Use, Regulations and Procedures (TEARAP). The purpose of this programme is to specifically train personnel involved in this kind of work. In addition, any experiment must count with the approval by the Health Products Regulatory Authority (HPRA) and the Animal Research Ethics Committee (AREC). Even though the process of obtaining such permits is time-consuming, I think they are absolutely necessary. Any biologist or veterinarian who works with animals must be very aware of the fact that they are working with live beings. If you are to perform any experiment on them, it is important to make sure that whatever you are doing is correctly done. Also, that the experimental design contemplates every possible factor in order to minimize both animal suffering and the number of individuals to be employed.

Finally, it is essential to guarantee the appropriateness of the animal model, in the absence of an in vitro one that may substitute for the in vivo procedure (“The 3Rs”). The ultimate goal is to obtain knowledge that cannot be obtained otherwise, and that can report certain benefit to society in the long term. In our field, the objective is to understand and improve everything related to infertility, both in species of livestock interest and in humans. Likewise, we try to apply that knowledge to improve the current techniques of assisted reproduction.

EM: During the last ESHRE congress in Barcelona, you presented the results on the effect of the environment of the bovine oviduct on embryonic development. This is a very interesting topic in order to understand the changes that occur in the embryo during the whole process. Could you tell us a bit about this piece of your research?

BR: In this study we wanted to see what happens when there is asynchrony between the stage of the embryo and the oviduct. Data on this topic are quite limited, probably due to the difficulty to access the oviduct in vivo. Thanks to the use of technologies like endoscopic transfer of embryos into the oviduct interesting results have been obtained. We have been able to study what happens when an embryo develops in a more advanced oviductal environment. Our results show that the asynchrony between the embryo and the oviduct exerts a negative impact on embryonic development.

EM: It seems that differences were found between the three groups of embryos used in the assay (day 4, day 7 and day 15). What is the main cause for these differences?

BR: Broadly speaking, we have observed that such asynchrony relates to a decrease in the survival and development of the embryo. However, embryo quality at the conceptus stage (day 15), assessed on its length, has been observed to be higher in asynchronous embryos. These observations had been previously reported by different research lines, where blastocysts had been transferred into advanced uteri. In these cases, exposure of embryos to an environment with higher concentration of progesterone seemed to accelerate their growth in those who survived. Therefore, there is a paradoxical effect: on one hand, a negative impact on development and degeneration; on the other hand, conceptuses are larger and presumably with better quality. We concluded that the overall effect is negative, since the group of cows that developed embryos in asynchrony displayed lower pregnancy rate (50%) than the group in synchrony (100% of them).

EM: So this could actually be a good method to understand the causes of miscarriages during the first weeks of gestation…

BR: Understanding the embryo-oviduct interaction gives us very valuable information. Despite the oocyte being normally fertilized, there are cases in which the embryo fails to implant in the uterus, so that pregnancy gets interrupted within the first two weeks after fertilization. There are many things to understand in order to diagnose and treat this type of subfertility.

EM: I guess it was complicated to plan all experimental steps in advance, along with the synchronization of the embryo with the oviduct…

BR: This type of work involves many complications at a technical level, and many people have been actively involved so that it could move forward. Here it is worth highlighting several people. First of all, my main supervisors: Dimitrios Rizos and Pat Lonergan, the minds behind all these experiments. Secondly, the magnificent laboratory equipment I have worked with, from the technicians to the rest of the graduate students and postdocs. I do not want my answer to become just acknowledgements, but I must mention someone else: José María Sánchez. He was a key part respecting the complex set up of the experiment.

EM: Could you tell us about future perspectives for your research line?

BR: There are still so many things to look into! Our ultimate goal is to continue deciphering embryo-maternal interactions, so that actual significant improvements on treatments and ART are feasible.

EM: Back to the ESHRE conference, you were one of the ESHRE Young Ambassadors who kept us posted via Twitter on many of the conferences and talks that took place. How was the experience?

BR: It was a wonderful experience! I think it was a very good opportunity to live the congress from another perspective, feeling you are part of it. It was also a great opportunity to meet many people in the field of reproduction. It is fortunate that ESHRE offers this option to everyone.

EM: How would you like to see Assisted Reproduction in the future? What are in your opinion the great challenges to overcome?

BR: I reckon there is a wide variety of things to be done, which is exciting! The first IVF baby was born just four decades ago; since then, over eight million children have been born all over the world thanks to ART. Modern advanced techniques have made this possible, and yet many couples fail to get pregnant after several assisted reproduction treatments. Even around 20% of patients who attend assisted reproduction treatments are diagnosed with “sterility of unknown origin”. This means there is still much to improve on diagnosis, and the same goes for treatments. Additionally, new and promising technologies are on the horizon, such as the gene-editing technology through CRISPR-Cas9.

EM: Thank you very much, Beatriz, for giving us this interview for Embryologist Media.

BR: Thank you for the opportunity!