Work performed:
Analysis of the biomechanics of the cell cluster and cell-cell communication. Further experiments on the relationship between the actomyosin ring and connexin/gap junction signalling were not as convincing as initially thought by the analysis of preliminary data. However, we found, even more interestingly, that ATP, a molecule never involved in neural crest migration before, has effect on synchronising cell cluster movements. This work package was therefore reduced in size and we started to focus on how the ATP controls coordination of the migrating cluster, without further characterisation of actomyosin ring.
Analysis of connexins/gap junctions and purinergic signalling. The work included expression analysis of ATP receptors and knockdown experiments in vitro and in vivo to study the function of these receptors and their involvement in response to ATP stimulation, dispersion effect and migration (including all appropriate controls), as well as expression analysis and knockdown experiments for connexins in vitro and in vivo (Milestone III). Furthermore, the effect of these knockouts on E-Cadherin expression levels was addressed.
Analysis of diffusible messenger and development of a zebrafish transgenic line to study calcium signals in the neural crest during migration in-vivo. The work involved analysis and characterisation of ATP-induced intracellular calcium signals and some of the inhibitors and analogues experiments (Milestone V). Characterisation of the effect of ATP stimulation (including apyrase experiments and all appropriate controls) on collective migration and coordination of the neural crests cluster, as well as effect on chemotaxis, motility and speed of neural crest cells both at the single-cell and collective level (Milestone V). Furthermore, the effect of ATP stimulation on E-Cadherin expression levels was addressed. Additionally, a zebrafish transgenic line was established to express the GCaMP5 calcium sensor specifically in the neural crest. This line was used to observe calcium transient patterns in vivo, using confocal imaging, in migrating neural crest cells in developing zebrafish embryos (Milestone IV). All experiments were carried out by the Researcher.
Result Dissemination:
Due to the early termination of the Fellowship, publication of the result in international scientific journals has not been achieved yet, however, the results obtained in the Fellowship months are of the highest relevance in the collective cell migration field, and future publication is still possible if other researchers in the lab will carry out the final needed experiments.
Besides participating in outreach activities, such as New Scientist Live 2016 London in collaboration with The Royal Society of Biology, I also set up a Twitter account of the Host Lab for the first time. This account had been well received by the scientific community and beyond, and it has quickly collected hundreds of followers in just a few months of being set up (
https://twitter.com/mayor_lab(se abrirá en una nueva ventana)). The page is being regularly updated by all members of the Host Lab to share interesting new research in the fields of cell and developmental biology, exciting new results and interesting discussion and presentations from the top scientists in the field during international conferences. Given the wide audience of this social media, this information is likely to reach not only scientists and collaborators, but also the general public. Evidence of this outreach potential is in several comments received from non-scientists to some posts on the page.
