As lockdowns ease and the prospect of attending a conference in person rises on the horizon, it might be time to take stock of what conferences offer that virtual meetings have fallen behind on: networking. Although networking is an important part of the conference experience, it can be daunting for some. I hope that these “Top ten tips” can help make networking a more enjoyable experience for those that aren’t feeling so confident. I don’t – by any means – consider myself an expert but I think that because I’ve actively had to work at my networking skills in my role as a Reviews Editor for Development, I’ve thought a lot about how to improve. I hope to share what I’ve learned through trial and error.
1. Cut yourself some slack
First, and most importantly, go easy on yourself! I can’t stress this enough. Big conferences can be overwhelming – even if you’re a natural communicator. It can be difficult to interact, especially when you are new to the field, jet-lagged, speaking a language that isn’t your mother tongue in a noisy room where other people might be speaking with unfamiliar accents. All these components can add up and it’s difficult! However, as with most things, the beginning is always the most challenging; it will get easier with time and better with practice.
2. Plan ahead
If you’re interested in speaking to someone in particular you can email them in advance of the meeting to arrange a time and place to meet. This way, you don’t need to worry about bumping into them by chance or spending time hunting them down. You can also use the conference Twitter hashtag, or a meeting app, to let other people know you’ll be at the meeting.
3. Start small
When meeting new people, don’t feel like every interaction needs to be revolutionary. For example, don’t feel pressure to start a 40-minute conversation about the minutiae of your research project, ending with a collaboration or a job offer. Set small goals. Sometimes, it’s enough just to introduce yourself to someone and catch up with them again later. Once they know who you are, you begin to lay the groundwork for a longer-term relationship. It’s enough to say who you are and that you’re looking forward to/enjoyed their talk. You can then leave it there for the time being.
4. The first impression
That being said, an introduction is quite important and it’s often the hardest thing to do. Personally, it’s something I still struggle with a lot and for many people with speech problems, such as stammers and stutters, saying your own name is often a trigger. Overall, my advice here is to practice and find something that works for you. If your introduction doesn’t go as planned, just carry on with the conversation; there’s no harm done, it just means you spend a bit of time correcting any misunderstanding. Don’t put pressure on yourself (remember Tip 1 above).
5. Keep it light
For me, one of the barriers to starting a conversation is a fear that I’ll be perceived as ignorant or stupid if I don’t know something or someone that I feel I should. This can be especially prevalent if you’re not confident about the scientific topic of the meeting. In these cases, make some small talk and find some common ground until you feel more comfortable. There are supposedly six degrees of separation between everyone in the world. I expect in scientific circles – developmental biology in particular – it’s probably half that. You’ll probably find it won’t take much time to find a colleague or institute in common.
Some fail-safe questions, which are obvious to some, include:
“Have you been to this meeting before?”
“Where are you coming from?”
“How was your journey here?”
“Do you get to spend some time visiting the local area?”
“What other meetings do you plan to go to?”
“Is anyone else from your group here?”
On the other hand, there are some questions I’d advise people to avoid, primarily about assuming someone’s academic position. I think it’s important to address our unconscious biases; even if you’re fairly confident of someone’s position (for example, if you’re attending a meeting for graduate students only), it’s still better to treat everyone equally.
Instead of…
Why not try…
“Are you a student/postdoc?”
“What is your background?”
“What do you work on?”
“What are you interested in?”
“What’s your project?”
“What are your research questions?”
“Who’s lab are you in?”
“Where are you based?”
“Who is your supervisor?”
“How long have you been there?”
6. Be genuine
I find developmental biology amazing and I can sometimes find myself getting a bit over excited. I used to worry that this would come across as being unprofessional but – as the cliché goes – be yourself. We are people first and scientists second. Commit to things that interest you and don’t feel apologetic for things you are (or aren’t) enthusiastic about.
7. Team up
Networking with a friend, colleague or even someone you met on the conference bus, can be a useful way of meeting new people without the pressure of trying to keep a conversation moving by yourself. You can ‘tag out’ and get some time to find your feet. Just make sure you don’t end up too reliant on support or only staying part of a group because you want to develop a unique identity and stay approachable (see Tip 8 below).
8. Be approachable
A reciprocal part of being a “good networker” is allowing others to network with you. Be conscious and aware of your surroundings; open the conversation circle when new people want to engage in your group discussion, or move aside if someone wants to listen in on a poster talk. Although travelling as a lab is great for bonding, and meeting old friends can be the personal highlight of a meeting, be careful not to come across as cliquey or exclusive. Talk to poster presenters and ask them questions – you never know what opportunities might arise out of a chance encounter.
9. Participate
Presenting a poster or a talk, or being a meeting organiser, is a great ice breaker and provides an opportunity for people to initiate a conversation with you.
10. Know when to call it a day
At the first opportunity for networking, it could be that you’ve been travelling halfway around the world and been awake for more than 24 hours. Know when to stop and get some rest; the harder the push yourself the more exhausted you will be and the harder it will become to keep up a conversation. Sometimes, knowing your limits is a really useful skill and there’s no shame in leaving the welcome drinks a little early.
Share your own wisdom
So, those are my suggestions! Do you have any other tips for networking? Share them in the comments below!
Semi-aquatic bugs (or Gerromorpha) likely derived from a terrestrial ancestor that evolved the ability to stand and move on the water-air interface about 200 million years ago. Access to previously unexploited ecological opportunities is associated with phenotypic evolution and often results in significant lineage diversification [2]. Our favorite bugs are no exception to the rule. Early-diverging lineages of semi-aquatic bugs occupy transitional zones and walk both on land and water, whereas derived lineages evolved rowing as a novel mode of locomotion on open-water surface [3,4]. Water surface invasion is commonly viewed as a stepwise process that involved both the diversification of leg morphologies and the evolution of densely arranged water-repellent ‘hairs’ that allow the insects to exploit surface tension. However, the mechanisms by which species develop traits adapted to the new ecological niche are not well understood.
Previous studies having predominantly focused on leg morphologies, we decided to explore the other side of the coin — the leg ‘hairs’. We first shown that the leg ‘hairs’ are nothing else than bristles [5]. Then the central question remains still on the table: how do semi-aquatic bugs evolve such a high density of bristles on their legs? Here started our investigation to find out what makes a water strider so hairy. After several years of leg-focused research, the Khila lab was quite excited and well determined to enter the bristle world.
Figure 1. The water strider Gerris buenoi resting on water surface. Image: courtesy of Abderrahman Khila.
Candidate gene approach
We first compiled a list of more than 120 genes known to play a role in bristle development in Drosophila, and searched for gene duplication events in the Gerromorpha. We found that the gene Beadex has two copies in the Gerromorpha. Whereas the copie BxA plays a role in bristle development in Gerris buenoi (= species with high bristle density), BxA is not significantly expressed during embryogenesis in Mesovelia mulsanti (= species with low bristle density). This result suggests that differences in leg bristle density might be partly attributable to differential expression level of BxA. We also identified two copies of the gene taxi in the semi-aquatic bugs that result from a duplication in the lineage leading to the Gerromorpha. We found that the gene taxiB evolves faster than taxiA and we detected positive selection along the taxiB lineage, suggesting functional divergence of the two copies. We depleted taxiA or taxiB transcripts using RNAi, and we observed defects in the development of leg bristles. However, we found that only ds-taxiB knockdown individuals exhibited shorter legs. Again, we were here to focus on the bristles, so we did not linger on the leg phenotype.
Genes involved leg bristle development in Gerromorpha…
By combining comparative transcriptomics and RNA interference, we identified six genes whose role in bristle development had never been documented in model organisms. We unraveled the role of the GPN-loop GTPase 2, the c-Myc binding protein MYCBP, and a protein-glutamate O-methyltransferase in bristle patterning; the role of the bHLH transcription factor Net and the MAP kinase signal responder protein Dodo in bristle elongation; and the role of the actin binding protein Simiate in bristle orientation and size.
… played also a role in leg growth!
Looking for bristle genes, finding bristle genes! It sounded like a pretty straightforward research project. Well, the truth turned out to be much more complicated and exciting. We found that suppressing the expression of bristle-related genes resulted in a leg shortening. This time, the leg phenotype was so predominant that we could not overlook it. We spent a substantial number of hours with our pictures of legs and our ruler. The quantification was unequivocal: most of the legs in the knockdown individuals were shorter. In brief, shorter and barer, or barer and shorter!
Cell division as a shared molecular mechanism
This hypothesis remained to be verified. To this end, we stained all nuclei with DAPI, and M-phase nuclei with anti-PH3 antibodies in the developing embryonic legs. We then compared the cell division rate between control and ds-taxiB knockdown individuals. We found that the legs T2, which are shorter in ds-taxiB individuals, showed a reduced mitotic activity. By doing so, we noticed that the orientation of cell division might also be affected. Thus, we measured the angle of division relative to the proximo-distal (PD) axis of the leg. We found that epithelial cells bias their orientation perpendicular to the PD axis in knockdown individuals, whereas these cells predominantly orient their divisions parallel to the PD axis in the control embryos.
Pleiotropy as a facilitator of diversification
The key message of the project began to take shape: leg length and bristle density, both necessary for water surface locomotion, are genetically correlated in the semi-aquatic bugs. We had the feeling we could go even a bit further. We plotted the bristle density against the leg length for controls and knockdown individuals of G. buenoi: linear correlation, checked! Because bristle density and leg length vary a lot across the semi-aquatic bugs, we extended our correlation analysis to all the species we had in the laboratory. The correlation between our two favorite traits still stands at the infraorder level.
The classical view of the origin of the semi-aquatic bugs implies the stepwise evolution of longer legs and denser leg bristles. Our findings suggest that the invasion of water surface might have been more ‘straightforward’ because of genetic pleiotropy. However, even if leg length and bristle density are both necessary for water surface locomotion, it does not necessarily mean these two traits have been selected for. Indeed, we cannot exclude that only one trait was selected whereas the other trait was simply a spandrel or by-product [6]. Our study represents the first step towards the deciphering of the molecular mechanism of high bristle density in the Gerromorpha, and further investigation will be needed to build a better picture of this ecologically relevant trait. Nevertheless, one thing is certain: in Gerromorpha, it is all about leg length!
[2] Schluter D. (2000). The ecology of adaptive radiation. Oxford University Press, Oxford.
[3] Andersen NM. (1976). A Comparative Study of Locomotion on the Water Surface in Semiaquatic Bugs (Insecta, Hemiptera, Gerromorpha). Vidensk. Meddel. Dansk Naturhist. Foren. Kjobenhavn 139: 337-396.
TEX13B is important for germ cell development and male fertility Umesh Kumar, Digumarthi V S Sudhakar, Nithyapriya Kumar, Hanuman T Kale, Rajan Kumar Jha, Nalini J Gupta, B N Chakravarthy, Mamata Deenadayal, Aarti Deenadayal Tolani, Swasti Raychaudhuri, P Chandra Shekar, Kumarasamy Thangaraj
Effects of α-crystallin gene knockout on zebrafish lens development Mason Posner, Kelly L. Murray, Brandon Andrew, Stuart Brdicka, Alexis Butterbaugh-Roberts, Kirstan Franklin, Adil Hussen, Taylor Kaye, Emmaline Kepp, Mathew S. McDonald, Tyler Snodgrass, Keith Zientek, Larry David
hPSC-Derived Enteric Ganglioids Model Human ENS Development and Function Homa Majd, Ryan M Samuel, Jonathan T Ramirez, Ali Kalantari, Kevin Barber, Zaniar Ghazizadeh, Angeline K Chemel, Andrius Cesiulis, Mikayla N Richter, Subhamoy Das, Matthew G Keefe, Jeffrey Wang, Rahul K Shiv, Conor J McCann, Samyukta Bhat, Matvei Khoroshkin, Johnny Yu, Tomasz J Nowakowski, Hani Goodarzi, Nikhil Thapar, Julia A Kaltschmidt, Faranak Fattahi
Vascular buds of Botryllus schlosseri from Ricci, et al.
Coupled myovascular expansion directs cardiac growth and regeneration Paige DeBenedittis, Anish Karpurapu, Albert Henry, Michael C. Thomas, Timothy J. McCord, Kyla Brezitski, Anil Prasad, Yoshihiko Kobayashi, Svati H. Shah, Christopher D. Kontos, Purushothama Rao Tata, R. Thomas Lumbers, Ravi Karra
Alteration of myocardial structure and function in RAF1-associated Noonan syndrome: Insights from cardiac disease modeling based on patient-derived iPSCs Saeideh Nakhaei-Rad, Farhad Bazgir, Julia Dahlmann, Alexandra Viktoria Busley, Marcel Buchholzer, Fereshteh Haghighi, Anne Schänzer, Andreas Hahn, Sebastian Kötter, Denny Schanze, Ruchika Anand, Florian Funk, Andrea Borchardt, Annette Vera Kronenbitter, Jürgen Scheller, Roland P. Piekorz, Andreas S. Reichert, Marianne Volleth, Matthew J. Wolf, Ion Cristian Cirstea, Bruce D. Gelb, Marco Tartaglia, Joachim Schmitt, Martina Krüger, Ingo Kutschka, Lukas Cyganek, Martin Zenker, George Kensah, Mohammad R. Ahmadian
Limb Specific Failure of Proliferation and Translation in the Mesenchyme Leads to Skeletal Defects in Diamond Blackfan Anemia Jimmy Hom, Theodoros Karnavas, Emily Hartman, Julien Papoin, Yuefeng Tang, Brian M. Dulmovits, Mushran Khan, Hiren Patel, Jedediah Bondy, Morris Edelman, Renaud Touraine, Geneviève Chanoz-Poulard, Gregory Ottenberg, Robert Maynard, Douglas J. Adams, Raymond F. Robledo, Daniel A Grande, Philippe Marambaud, Betsy J Barnes, Sébastien Durand, Anupama Narla, Steven Ellis, Leonard I. Zon, Luanne L. Peters, Lydie Da Costa, Jeffrey M. Lipton, Cheryl L. Ackert-Bicknell, Lionel Blanc
Myeloid-biased HSC require Semaphorin 4A from the bone marrow niche for self-renewal under stress and life-long persistence Dorsa Toghani, Sharon Zeng, Elmir Mahammadov, Edie I. Crosse, Negar Seyedhassantehrani, Christian Burns, David Gravano, Stefan Radtke, Hans-Peter Kiem, Sonia Rodriguez, Nadia Carlesso, Amogh Pradeep, Nicola K. Wilson, Sarah J. Kinston, Berthold Göttgens, Claus Nerlov, Eric Pietras, Marion Mesnieres, Christa Maes, Atsushi Kumanogoh, Thomas Worzfeld, Peter Kharchenko, David T. Scadden, Antonio Scialdone, Joel A Spencer, Lev Silberstein
Developmental origins of cell heterogeneity in the human lung Alexandros Sountoulidis, Sergio Marco Salas, Emelie Braun, Christophe Avenel, Joseph Bergenstråhle, Marco Vicari, Paulo Czarnewski, Jonas Theelke, Andreas Liontos, Xesus Abalo, Žaneta Andrusivová, Michaela Asp, Xiaofei Li, Lijuan Hu, Sanem Sariyar, Anna Martinez Casals, Burcu Ayoglu, Alexandra Firsova, Jakob Michaëlsson, Emma Lundberg, Carolina Wählby, Erik Sundström, Sten Linnarsson, Joakim Lundeberg, Mats Nilsson, Christos Samakovlis
A remote lecture series roadmap to equity, diversity, and inclusion in STEM Evan A. Boyle, Gabriela Goldberg, Jonathan C. Schmok, Jillybeth Burgado, Fabiana Izidro Layng, Hannah A. Grunwald, Kylie M. Balotin, Michael S. Cuoco, Keng-Chi Chang, Gertrude Ecklu-Mensah, Aleena K. S. Arakaki, Noorsher Ahmed, Ximena Garcia Arceo, Pratibha Jagannatha, Jonathan Pekar, Mallika Iyer, DASL Alliance, Gene W. Yeo
Virtually the same? Evaluating the effectiveness of remote undergraduate research experiences Riley A. Hess, Olivia A. Erickson, Rebecca B. Cole, Jared M. Isaacs, Silvia Alvarez-Clare, Jonathan Arnold, Allison Augustus-Wallace, Joseph C. Ayoob, Alan Berkowitz, Janet Branchaw, Kevin R. Burgio, Charles H. Cannon, Ruben Michael Ceballos, C. Sarah Cohen, Hilary Coller, Jane Disney, Van A. Doze, Margaret J. Eggers, Edwin L. Ferguson, Jeffrey J. Gray, Jean T. Greenberg, Alexander Hoffmann, Danielle Jensen-Ryan, Robert M. Kao, Alex C. Keene, Johanna E. Kowalko, Steven A. Lopez, Camille Mathis, Mona Minkara, Courtney J. Murren, Mary Jo Ondrechen, Patricia Ordoñez, Anne Osano, Elizabeth Padilla-Crespo, Soubantika Palchoudhury, Hong Qin, Juan Ramírez-Lugo, Jennifer Reithel, Colin A. Shaw, Amber Smith, Rosemary J. Smith, Fern Tsien, Erin L. Dolan
One of the key objectives of the Node Network is to allow scientists, especially early-career researchers (ECRs), to raise their profiles in the developmental and stem cell biology community. With this in mind, and as part of our second birthday celebrations, we are delighted to launch our discussion and networking event, ‘Promoting yourself as an ECR’ hosted with our sister community sites FocalPlane and preLights. The interactive event will begin with a panel discussion with our three invited panellists Maria Abou Chakra, Pablo Sáez and Sarvenaz Sarabipour and then continue with a networking event where you can meet the panellists, representatives from The Company of Biologists and other ECRs.
Dr Maria Abou Chakra is a research associate at the University of Toronto, where her research focuses on mathematical modelling of stem cell development. She is the organiser of the Modelling Cell Development & Regeneration Discussion Group, as well as having been involved in outreach, mentorship and EDI events.
Professor Pablo J. Sáez is a new PI at UKE, Hamburg, where his international team is studying the role of cell communication and migration, with a particular focus on immune cells. Pablo is a regular contributor to our preprint posts on FocalPlane, and is an advocate for better representation for ECRs in academic conferences.
Dr Sarvenaz Sarabipour is an assistant research scientist at Johns Hopkins University, where her research focuses on receptor signalling networks at the cell and tissue level. As well as her scientific interests, she is an active advocate for ECRs, open science, mentorship, and diversity in science.
Illustrations from On Growth And Form by D’Arcy Thompson
Whereas previously, most biologists felt that the living world had to be dealt with in a completely separate way from the non-living world, D’Arcy was saying, “Actually, no. We can look at a lot of basic forms in biology and explain their growth and shapes according to fundamental laws of physics and mathematics. A living thing may grow in exactly the same way as a non-living thing because it’s just obeying these fundamental laws.”
Matthew Jarron describing D’Arcy Thompson
In the latest episode of the Genetics Unzipped podcast, we’re exploring the life and work of D’Arcy Wentworth Thompson – one of the first scientists to bring together the worlds of mathematics and biology in the quest to understand how living things are built. Dr Kat Arney sat down with the curator of the University of Dundee’s D’Arcy Thompson Zoology Museum, Matthew Jarron, to find out more about this larger than life character…
If you enjoy the show, please do rate and review on Apple podcasts and help to spread the word on social media. And you can always send feedback and suggestions for future episodes and guests to podcast@geneticsunzipped.com Follow us on Twitter – @geneticsunzip
In skeletal muscle, we know that two stem/progenitors, muscle satellite cells (MuSCs) and mesenchymal progenitors (also known as FAPs), are critical for homeostasis, regeneration, and growth including muscle hypertrophy. Although the involvement of MuSCs in muscle hypertrophy has been well studied, the mechanism underling MuSC proliferation had not been investigated. One reason for this omission was that MuSC proliferation was thought to occur by the same mechanism as the well-studied process of muscle regeneration and muscle loading. In addition, the roles of mesenchymal progenitors in muscle hypertrophy had not been elucidated. Based on this background, we started this project and recently published results (Kaneshige et al., Cell Stem Cell, 2022) that shed light on the mechanisms regulating MuSC proliferation by mesenchymal progenitors in a surgical loaded muscle. In this paper, we presented data that showed 1) critical roles of mesenchymal progenitors in MuSC proliferation, 2) involvement of Yap1/Taz in loaded mesenchymal progenitors, 3) the contribution of mesenchymal progenitors-derived Thrombospondin-1 (Thbs1) as the target of Yap1/Taz to MuSC proliferation, and 4) Thbs1-derived peptide induces the proliferation of Calcitonin receptor (CalcR)-mutant MuSCs. However, the results were not obtained in the order shown in the figures; for example, the analysis of Pa-Yap1/Taz was the last experiment of the main project. In this blog, the results will be presented in the order in which they were discovered.
Depletion of mesenchymal progenitors in loaded muscle
In our previous project (Fukuda et al., 2019), we had noticed morphological changes in mesenchymal progenitors by mechanical load using FACS analyses (actually, we did not put a high priority on it due to our focusing on MuSCs at that time…). These data and our new hypothesis, that mesenchymal progenitors might be involved in muscle hypertrophy, encouraged us to initiate this project. Fortunately, our team (Professor Uezumi, co-corresponding author) had already established an experimental model for depleting mesenchymal progenitors. We started to collaborate before his first publication using Pa-DTA (PdgfraCreERT::Rosa DTA) mice (Uezumi et al., 2021). We were very excited when our data convinced us that the expansion of MuSCs during overload was significantly reduced in Pa-DTA mice depleted mesenchymal progenitors (Kaneshige et al., 2021).
Microarray analysis of mesenchymal progenitors
How do mesenchymal progenitors regulate MuSCs in loaded muscles? To address this crucial question, we conducted microarray analysis of mesenchymal progenitors from loaded muscle and explored MuSC regulators. We focused on genes with the following three features; 1) upregulated by mechanical loading, 2) secreted molecules, 3) different expression pattern from muscle injury model. We finally got four candidates including Thbs1. Since the gene expression analysis of Pa-Yap1/Taz-cdKO, which will be described later, was performed by RNA-seq, we re-analyzed normal mesenchymal progenitors by RNA-seq for consistency.
Screening of MuSCs regulator candidates
To uncover a signaling mechanism between mesenchymal progenitors and MuSCs, we conducted an in vivo screening with inhibitory antibody against the above candidates or their receptor. For Thbs1, we tested an antibody against CD47, which is one of well-known receptor of Thbs1 and highly expressed in MuSCs. As all antibodies were commercially available, this was a feasible strategy. Thus, we analyzed the loaded muscles and found that new myonuclei supply was reduced in mice treated with anti-CD47 antibody.
Injection of PKHB1 into CalcR-cKO
Elucidation of the role of CalcR in MuSCs has been one of the main themes in our laboratory (Baghdadi et al., 2018; Fukada et al., 2007; Yamaguchi et al., 2015; Zhang et al., 2019). CalcR-mutated MuSCs exhibit the increased expression of cell cycle-related genes including Ki67. However, no substantial cell division was observed. In contrast, MuSC number was decreased in time dependent manner after CalcR-depletion by tamoxifen injection.
Early on this project, we had observed that CalcR expression in MuSCs were decreased in the surgical loaded muscle, however, no further analysis had been conducted. After that, we found that PKHB1 (CD47 agonist peptide designed based on CD47 binding sequence of Thbs1) promoted MuSC proliferation in loaded muscle, but not in non-loaded muscle. To know the effect of CalcR on MuSC expansion by PKHB1, we tried to inject PKHB1 into sedentary CalcR-cKO mice without surgical muscle overload. In these analyses, MuSC number on isolated myofibers from the treated mice were counted in blind manner. We were very surprised to see that the number of MuSCs in CalcR-cKO was increased by PKHB1 injection. We also observed the MuSCs-derived new myonuclei which were rarely detected in untreated CalcR-cKO mice.
Yap1/Taz in mesenchymal progenitors
As the downstream target of CalcR signaling, we had previously focused on Yap1 (Zhang et al., 2019), therefore, we already had Yap1-floxed mice. This meant that it was easy for us to generate PdgfraCreER::Yap1flox/flox mice. As Yap1 is known as a mechanical transducer, we expected that the surgical loaded model would be nice way to know the role of Yap1 in mesenchymal progenitors. However, we could not detect consistent difference between control and Pa-Yap1 (PdgfraCreERT::Yap1flox/flox). Therefore, by collaborating with Professor Potent, Dr Watanabe, and Professor Braun, we generated Pa-Y/T-cdKO (PdgfraCreERT::Yap1flox/flox::Tazflox/flox) mice, and could observe the remarkable differences between control and Pa-Y/T-cdKO mice.
RNA-seq analyses links Yap1/Taz and Thbs1
As mentioned above, we had already obtained the data indicating that the blockage of CD47 inhibited the increased number of MuSCs-derived myonuclei and that the CD47 agonist, PKHB1, promoted MuSCs proliferation in vivo. As CD47 expression is ubiquitously expressed in many types of cells, we needed to use MuSCs-specific CD47-depletion mice. Fortunately, Professor Matoba and Dr Saito kindly provided us CD47-floxed mice, which enabled us to analyze MuSC-specific CD47-cKO (Pax7CreERT2::Cd47 flox/flox) mice.
In collaboration with Professor Ohkawa and Dr Maehara, we performed RNA-seq analysis of Pa-Y/T-cdKO mesenchymal progenitors and our data demonstrated reduced expression of Thbs1 as well as Yap-target genes. By performing some additional experiments including nice immunostaining results from Professor Uezumi’s team, we were sure that Thbs1 is one critical target by Yap1/Taz in loaded mesenchymal progenitors and promotes MuSCs proliferation via CD47.
In the end, we are really grateful to all authors involved in this project. Sometimes, the research object already exists closed to you, or sometimes, initial data has already been in your hand, which might be serendipity. We already had HeyL-deficient mice which provided the basis of this project (Fukuda et al., 2019). Kobe University, where CD47-floxed mice were housed in Professor Matoda Lab., is geographically very close to Osaka University. This project taught us that critical materials or animals could already be close to us and that new collaborations can provide unexpected links between different projects.
References
Baghdadi, M.B., Castel, D., Machado, L., Fukada, S.I., Birk, D.E., Relaix, F., Tajbakhsh, S., and Mourikis, P. (2018). Reciprocal signalling by Notch-Collagen V-CALCR retains muscle stem cells in their niche. Nature 557, 714-718.
Fukada, S., Uezumi, A., Ikemoto, M., Masuda, S., Segawa, M., Tanimura, N., Yamamoto, H., Miyagoe-Suzuki, Y., and Takeda, S. (2007). Molecular signature of quiescent satellite cells in adult skeletal muscle. Stem Cells 25, 2448-2459.
Fukuda, S., Kaneshige, A., Kaji, T., Noguchi, Y.T., Takemoto, Y., Zhang, L., Tsujikawa, K., Kokubo, H., Uezumi, A., Maehara, K., et al. (2019). Sustained expression of HeyL is critical for the proliferation of muscle stem cells in overloaded muscle. Elife 8, e48284.
Kaneshige, A., Kaji, T., Zhang, L., Saito, H., Nakamura, A., Kurosawa, T., Ikemoto-Uezumi, M., Tsujikawa, K., Seno, S., Hori, M., et al. (2022). Relayed signaling between mesenchymal progenitors and muscle stem cells ensures adaptive stem cell response to increased mechanical load. Cell Stem Cell 29, 265-280 e266.
Uezumi, A., Ikemoto-Uezumi, M., Zhou, H., Kurosawa, T., Yoshimoto, Y., Nakatani, M., Hitachi, K., Yamaguchi, H., Wakatsuki, S., Araki, T., et al. (2021). Mesenchymal Bmp3b expression maintains skeletal muscle integrity and decreases in age-related sarcopenia. J Clin Invest 131, e139617.
Yamaguchi, M., Watanabe, Y., Ohtani, T., Uezumi, A., Mikami, N., Nakamura, M., Sato, T., Ikawa, M., Hoshino, M., Tsuchida, K., et al. (2015). Calcitonin Receptor Signaling Inhibits Muscle Stem Cells from Escaping the Quiescent State and the Niche. Cell reports 13, 302-314.
Zhang, L., Noguchi, Y.T., Nakayama, H., Kaji, T., Tsujikawa, K., Ikemoto-Uezumi, M., Uezumi, A., Okada, Y., Doi, T., Watanabe, S., et al. (2019). The CalcR-PKA-Yap1 Axis Is Critical for Maintaining Quiescence in Muscle Stem Cells. Cell reports 29, 2154-2163 e2155.
Plant Postdocs started the journey as an organization in September 2019, a few months before the pandemic. At the beginning, our goal was to organize virtual webinars, seminars, and career sessions, where postdocs could share their works and sessions about job opportunities and professional development. As a small start-up organization with leadership team members from different institutions, internet was the only way for us to reach our peers. But, back in our mind we knew that it is challenging to organize and run every event online. People are not very used to the idea of virtual events, and most importantly not properly equipped and comfortable.
At the beginning of the 2020, the COVID-19 spread forced universities and institutes to go online and cancel in-person classes and seminars. Even before the lockdown was announced in February, we scheduled our first two virtual seminars. By March 2020, when we had our first webinar, serendipitously it was the first few weeks of pandemic lockdown. To our surprise, the timing for a virtual webinar could not be better than that. That was the beginning of the virtual webinar series of Plant Postdocs. We had a really good participation and feedback from the community, but we also realized there is room to improve, a better way to reach our peers, and engage the community.
Flyer of the 2021 Fall semester career webinar organized by Plant Postdocs
Besides the events organized in each month or semester, we created the day to day communication platform on Slack. Slack is very organized to keep our conversation focused in different channels. Like any other virtual platform, Slack took time to become a popular way of communication among scientific society. Plant Postdocs’ experience is no different in this case. Compared to our official Twitter platform (which has more than 5k followers), we have currently 300+ members on Slack. We post our activities on both platforms, Twitter and Slack; and we do not want anyone to miss important events and updates just because they are not present or active in one of those platforms. In addition, we have an autobot in our Slack as a channel, where all of our tweets appear automatically. Although the group is smaller in Slack, so far this is the best way to do the community conversation and pin the link of important resources as well.
Since the beginning to recent days, unfortunately we are not completely out of COVID-19 era. This practically means no in-person conferences for two academic years. However, the Plant Postdocs’ webinars were an opportunity for postdocs to present their work when in-person conferences were canceled. The cancellation of in-person events led people to adapt better for the virtual events. Over the time, we have observed that panelists and participants are more engaged and comfortable in virtual events. Even as a regular organizer, we find it easier to organize events these days compared to the beginning.
Recorded virtual webinars and seminars in the Plant Postdocs’ YouTube channel
Virtual events helped us to keep going through the difficult pandemic lockdown time. At the same time, everyone’s schedule was full of multiple Zoom events in a day. Too many virtual events caused “Zoom fatigue” in the meantime. As a matter of fact, most virtual events started to observe less and less participants over the time. Plant Postdocs’ events were no exception in this case. To overcome the declining participants, we have decided to diversify our webinar topics.
Our organization is dedicated to postdocs in the plant biology field. At the beginning, we used to organize two career webinars, one about academic jobs and another for industry/Government jobs, in each semester. Along with Zoom fatigue, highlighting similar topics in the webinars again and again made it monotonous to our regular participants. To overcome this issue, we organized webinars on scientific writing and editing, preparation for postdoctoral fellowship. At the same time, we started to invite panelists from start-up biotech companies, Assistant professors from PUIs (Primarily Undergraduate Institutions), and journal editors. Diversifying the webinar topics and broadening the panelists helped us to regain the participants in our virtual webinars.
As a scientist, we perform experiments every day and learn through our mistakes and experiences. Organizing virtual events is a similar experience for Plant Postdocs as an organization. Over the two and half years, Plant Postdocs not only provided resources for the community, but also created a model system to follow and retain participants in this evolving era of virtual scientific events.
We believe the story of Plant Postdocs is an ideal example for non-profit scientific organization in this evolving era of science communication. We are committed to serve our community in future by providing resources, such as workshops, webinars, and training for ECRs, creating a platform for postdocs to present their work in our monthly seminar series, promoting pre-print and PREreview, and making the platform accessible to everyone.
During embryonic development, bone formation begins with the condensation of mesenchymal stem cells (MSCs). In a few places of our body, such as in the skull and the shoulder blades, mesenchymal condensations differentiate directly into bone-forming cells called osteoblasts. These osteoblasts make bones by directly laying down bone matrix, and this process is known as intramembranous ossification. In most other condensations in the embryo, such as those that eventually became the arms and legs, bones are formed by a different process called endochondral ossification, where a cartilage mold is first formed by chondrocytes, which is then replaced by the incoming osteoblasts, laying down bone matrix using the cartilage scaffold. Therefore, osteoblast function plays an important role in both types of ossification in our body and have clinical implications in both skeletal development and diseases like osteogenesis imperfecta and osteoporosis.
Runx2 and Osterix (also known as Sp7) are two of the most well-studied transcription factors that control osteoblast differentiation. Runx2 belongs to the Runx family that is composed of three genes, Runx1/Cbfa2, Runx2/Cbfa1, and Runx3/Cbfa3. Runx2 and 3 together are essential for chondrocyte maturation (1) and Runx2 is important also for osteoblast differentiation, as demonstrated by the complete lack of ossification in Runx2 knockout mice (2, 3). The molecular mechanisms by which Runx2 regulate osteoblast differentiation has been elucidated. All Runx2 family proteins contain a DNA-binding runt domain. The Runx proteins form heterodimers with transcriptional co-activator core binding factor b (Cbfb) in vitro (4) and specifically recognize a consensus sequence, PyGPyGGTPy (5), to upregulate a variety of osteoblast lineage-specific genes, such as Sp7 (osterix), Ocn (osteocalcin), and Bsp (bone sialoprotein) (6, 7). Similar to Runx2, Sp7 knockout mice also demonstrated lack of ossification. But in contrast to Runx2 knockout, the Sp7 knockout mice do express Runx2 in osteogenic cells, thus suggesting Sp7 acts downstream of Runx2 during osteoblast differentiation (8).
Over the years, the role of Sp7 in osteoblast function has been confirmed in humans. Pathogenic variants in SP7 have been described in patients with recessive osteogenesis imperfecta (OI type XII), which is characterized by generalized osteoporosis (9), and genome-wide association studies (GWAS) have identified common genetic variants in SP7 to be associated with bone mineral density in the general population (10). However, despite the physiological and clinical importance of SP7, much is yet to be learnt about its molecular mechanisms of action. Based on its similarities with other SP family members, SP7 was initially thought to bind GC-rich sequences (11). However, other studies have shown evidence for a lack of such binding preference for GC-rich sequences (12). In 2016, a study from Andrew McMahon’s group showed that SP7 differs from other SP proteins in that it has a lower binding affinity for GC-rich sequences, and instead preferentially binds to AT-rich sequences in osteoblast target genes through interactions with DLX proteins (13). These findings provided important insights into why SP7, but not other SP family proteins with similar zinc finger domains, is uniquely important for osteoblast function. However, these findings were based on in vitro chromatin immunoprecipitation sequencing (ChIP-Seq) using osteoblasts, and thus it remains unclear whether this unique AT-motif-binding of SP7 is important in vivo, as at least until a human subject is found to have such binding preference disrupted.
And that brings us to our work recently published in Nature Communications (14). In our latest study, we presented an Austrian boy with a complex skeletal disease which included craniosynostosis, severe scoliosis, long bone fragility, with areas of increased bone, particularly with thickened intramembranous bones, and other areas of decreased bone. In that patient, we identified a pathogenic heterozygous missense variant in SP7 (S309W). The complex skeletal phenotype and the apparently dominant nature of the variant differ markedly from the prior cases of SP7-associated recessive osteogenesis imperfecta and is not readily explained by a simple loss of SP7 function and osteoblast formation.
We created a mouse with the orthologous missense variant in Sp7, which partially recapitulated the human skeletal phenotype. Importantly, we showed evidence that the variant altered the binding specificity of SP7, with increased binding to GC-consensus sequences and decreased binding DLX proteins and to AT-rich motif (thus a neomorphic/gain-of-new-function mutation rather than a simple loss- or gain-of-function mutation). The variant tends to reverse the unique sequence specificity of SP7, causing it to revert to a specificity more similar to the other SP family members. Our current study therefore provides the first in vivo evidence that the unique AT-binding specificity of SP7 is indeed essential for normal bone development in vivo. Importantly, our study also suggests the possibility that other unresolved rare genetic disorders could also be caused by neomorphic mutations in transcriptional regulators.
The Quintay Practical Course in Developmental Biology 2020 student cohort interviewed Dr. Angela Nieto Toledano
Dr. Angela Nieto is a researcher, professor, scientific communicator whose work builds bridges between embryology and health research. Angela is the president of the ISDB (International Society of Developmental Biology), and she has received several scientific awards in recognition to her outstanding work – including her recent incorporation to the Royal Academy of Exact, Physical and Natural Sciences of Spain.
During the last MBL course in Quintay (January 2020), 18 students from the field of developmental biology had the opportunity to share meals, lectures, talks, ideas and ask advice from Dr. Nieto. Some of the discussions are summarized in this interview to share the experience more broadly with the developmental biology community.
Dr. Angela Nieto and Dr. Roberto Mayor
1. What is snail and how did you start studying this gene? Why is snail so important for development and at the same time for processes involved in the evolution of cancer?
During my postdoc in London I had been working with David Wilkinson on the search for genes important for the development of the brain. Just before returning to Spain at the end of 1992, I found a new gene that was expressed in cells that were moving from one place to another in the embryo, a well-known process for which there was not much information on how it happened. We called that gene Slug, now known as Snail2, as we had previously isolated another family member, Snail1. Just then I decided that, in my new lab in Spain, I would devote my efforts to finding out how cells in the embryo move to form organs. In 1994, we found that Snail2 downregulation in chicken embryos prevented migration of both the neural crest and the mesoderm from the neural tube and the primitive streak, respectively. In this paper, published in Science, we hypothesized that the pathological activation of these genes could be involved in the acquisition of migratory and invasive properties in cancer cells, as at the cellular level, the delamination of cells from the primary tumour to start the metastatic cascade looked very similar to the delamination of neural crest cells from the neural tube. In 2000, we could show that this was the case.
2. What other things has your lab been interested in over the years?
We have continued to study the mechanisms that drive morphogenesis. Over the years we have found that the mechanisms that should only be active in the embryo are activated again in an aberrant way in different diseases, including cancer or organ degeneration. We continue to learn from the embryos in order to better understand the diseases and propose better therapeutic strategies.
3. Do you think that, in addition to effort and good ideas, there are rules to achieve success in a project? What are yours?
Passion, enthusiasm and resilience. It is also important to mention that scientific activity has degrees of freedom that other jobs do not have and this is very satisfying.
4. Your laboratory, like many in today’s world, is made up of researchers and students from multiple backgrounds. What are the advantages of this diversity?
In the lab, like in other aspects of our life and in nature in general, diversity is a source of richness and a selective advantage. A multicultural environment fosters knowledge and generosity and, in any case, science is universal and should promote strong links among individuals. We are now in the middle of a devastating and unexpected crisis, and we know that we will overcome Covid-19 only working all together. Scientific research has now taken centre stage in the news, let´s hope that it will continue to be, firstly because scientists could find a treatment and an efficient vaccine, and secondly because society realized the importance of investing in science as the only way to secure a prosperous future.
5. What advice would you give to female young researchers who want to develop a prolific career?
The career of a scientist is not easy, but in the case of women it is even more complicated because the crucial stage when the future is decided coincides with that of motherhood. We are still a long way from women being able to reconcile both aspects naturally. On the other hand, there are fields of science that have been and still are associated with men, and this means that many girls do not have female role models to look at. Girls develop the so called “dream gap”, something like the conviction that they are not going to be able to do well in scientific disciplines. This is now more dramatic than ever, as we are in a world of Technology, Artificial Intelligence and “Big Data”, where it is predicted that 80% of future jobs will be in the STEM disciplines. Therefore, we must help in two ways. The first is to help women who want to be scientists to continue their careers while they are mothers and the second is to change the sexist education that girls receive from the beginning. To do this, we must show models of successful women so that they see that it is possible and that they can do it too. In my case, I have been very lucky because I have always had the support of my family and my partner, and I have not had episodes of discrimination as I have progressed in my career. But we still have a long way to go.
6. Finally, this international course is composed mostly of students from Latin America, where many people dream of working in science since elementary school but have not had the best opportunities. What is your message to them?
First of all, I have to say that I have been very much impressed by the knowledge and enthusiasm of the students, and it has been a real pleasure to spend these days in Quintay. Thus, I want to thank Roberto Mayor and the other organizers for their efforts in organizing such a wonderful course for the last 20 years (and the many to come). My message to the students is quite simple, think high and work hard. And also think that science is fun and that, little by little, maybe we can help to better understand disease and improve our future.
The next EMBO Practical Course on Developmental Biology in Quintay, Chile from 4-16 January 2023 is now accepting applications. You can find more information on the course and how to submit your application here.
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