The Buckley group strives to understand how mechanics and biochemical signalling work together to shape epithelial organs during morphogenesis. Our long-term goal is to identify mechanisms by which a dysregulation of this balance contributes to diseases. We are currently particularly interested in understanding the links between apical-basal cell polarity, cell-cell adhesion, and cellular mechanics during the process of vertebrate secondary neurulation and mammalian early epiblast formation. To address these questions, we use in vivo optogenetic approaches to manipulate actomyosin contractility deep within the zebrafish brain. This has demonstrated that forces can asymmetrically propagate across large tissue scales during neural tube morphogenesis. We also use multicellular stem cell culture models of epiblast formation, with which we have uncovered a novel, Cadherin-mediated mechanism of apical membrane initiation site localisation during epithelial organ development.

My undergraduate degree was in Biological Sciences at Oxford in 2005. During my PhD I developed models of myelination in zebrafish with Robin Franklin and DanioLabs at Cambridge in 2009. I became obsessed with apicobasal polarity and epithelial tubes during my postdoc with Jon Clarke at KCL until 2017 and hooked on optogenetics during a visiting scholarship to Orion Weiner’s laboratory at UCSF in 2014. In 2015 I also started another exciting role; parenting, and took a year’s parental leave, following by working part-time. I started my lab at the department of Physiology, Development and Neuroscience at Cambridge with a RS Dorothy Hodgkin Research and a WT/RS Sir Henry Dale Fellowship in 2018. In 2019 I started another parenting role, this time taking 6 month’s leave and working part time for several years. I moved to the division of Molecular & Cellular Function at the University of Manchester as a senior research fellow in 2024. My lab studies the interrelationship between mechanics, signalling and morphogenesis within epithelial organs.

How do single cells assemble themselves into highly organized and complex structures such as organs?

Developmental processes are highly interrelated. The behaviour and fate of a single cell will depend on its chemical and mechanical environment and these factors also operate across very different length scales. My lab is tackling how mechanics and signalling work together during morphogenesis. In the longer term, we are interested in understanding how are the fundamental mechanisms behind 3D tissue organization relevant to disease? and can we manipulate these mechanisms for therapeutics and bioengineering?

An interesting developmental process in which to study the fundamental mechanisms of organ formation is the genesis of epithelial tubes and cavities, which make up the precursors to many organs in the body (such as the brain). These are made from polarised epithelial cells. Interestingly, there is more than one way in which these structures can arise, even within the same tissue. For example, the anterior neural tube of the amniote arises via folding of the already polarised neural ectoderm. This is called primary neurulation and is a well -studied process, driven by mechanisms such as actomyosin-mediated contractility at the apical ends of the cells. However, if you look at posterior neural tube within the same organism, it arises via a very different mechanism. In this case, cells within the neural rod undergo a mesenchymal to epithelial transition (MET) shape change inside a solid organ primordium and the lumen then opens via hollowing. Little is known about the biochemical or mechanical mechanisms driving secondary neurulation or why amniotes adopt these two different modes of neural tube formation.  However, there is a high incidence of neural tube defects arising from the intermediate region between primary and secondary neurulation. Understanding the mechanisms driving secondary ‘opening’ tubes vs primary ‘closing’ tubes is therefore an important research goal.

In addition to furthering our understanding of secondary neurulation and its associated defects, studying the fundamental mechanisms behind polarisation and lumenogenesis of hollowing tubes also has widespread relevance during the development of many organs (e.g. kidney nephrons, gut), as well as the early embryo morphogenesis of the epiblast. The physiological function of all of these structures relies on a centrally located and continuous fluid filled lumen. However, we still don’t know much about how these partially mesenchymal, solid structures transform into highly polarised, and then OPEN epithelial tubes and cavities.

To understand how an epithelial tube is built, we are investigating

1. How do epithelial tubes polarise in the correct place?

Our lab recently used a mouse embryonic stem cell culture model of the mammalian epiblast to identify a new Cadherin-mediated mechanism of apical membrane initiation site (AMIS) localisation within developing epithelial cavities. This suggests that cell-cell adhesion acts as the first symmetry-breaking event in de novo polarising epithelial tubes and cavities (Liang et al., 2022). This is important because it provides a mechanism by which single polarising cells localise their apical membrane in relation to their neighbours. This enables a coherent epithelial tissue to arise from a dynamic population of dividing and moving progenitor cells. However, it is also accepted in the literature that cell division is sufficient to direct AMIS localisation, via docking of apical proteins at the mitotic midbody (reviewed in Buckley & St Johnston 2022). Other collaborative work within the zebrafish neural tube suggests that this is likely due to the mitosis-dependent alignment of cytokinetic midbodies with Cadherin and apical polarity proteins (Symonds & Buckley et al., 2020). We are now interested in understanding the role of actomyosin flows in mediating this alignment, and in further understanding the Cadherin-dependent mechanism of AMIS positioning.

2. How do hollowing epithelial tubes open?

Our lab uses zebrafish neurulation as a tractable in vivo experimental model of secondary-like neurulation. The zebrafish hindbrain and spinal cord primordium forms via de novo polarisation and hollowing. Zebrafish embryos also develop rapidly and are amenable to high-resolution live imaging from subcellular to tissue scales, as well as small-molecule chemical manipulation of signalling via immersion. Whilst the role of contractile actomyosin at the apical lateral junctions of primary folding neural tubes is well established, we have recently used the zebrafish neural tube to investigate the role of actomyosin contractility during secondary-like (opening) neurulation. We have found that, despite the different modality of lumenogenesis, actomyosin contractility is still required for the neural tube lumen inflation, but appears to be operating via a different subcellular mechanism, which we are currently investigating. 

3. How is morphogenesis coordinated along the anterior posterior axis?

To directly study the interrelationship between mechanics and biochemical signalling during morphogenesis of epithelial tubes, we have developed in vivo optogenetic approaches, which allow us to image the behaviour of cells and tissue before, during and after a precise and reversible manipulation in cell polarity or cell signalling deep within the developing zebrafish neural tube (Buckley et al., 2016, Buckley 2019, Crellin and Buckley 2023). We have recently used this approach to locally activate actomyosin contractility along the medio-lateral axis within the zebrafish neural rod and then to measure the response of surrounding tissue in response to a local increase or relaxation of cell stress. This has uncovered that a rapid and long-ranging, elastic force propagation, suggesting active force propagation within the tissue. Interestingly, this response was highly asymmetric, transmitting in either the anterior or posterior direction. We are currently interested in the potential underlying biases in the tissue that might explain this asymmetry, and how cell signalling is affected by these long-ranging forces.

I have expertise in vivo live imaging and optogenetics within zebrafish embryos and am happy to collaborate in furthering these techniques.

I have teaching experience across a range of courses from my time at the University of Cambridge. I taught developmental and regenerative neuroscience to 1^st and 2^nd year medics and vets and introduced new 3^rd year lectures on neurulation, apical- basal polarity and epithelial morphogenesis. I also have extensive neuroanatomy demonstration experience. I enjoy bringing biophysical experimental and analytical approaches to researching morphogenesis from our lab into the classroom, both at the University and at international EMBL and MBL workshops.

ED&I

During my career I have taken two periods of parental leave and worked part-time for several years. Having experienced the challenges of bringing up small children whilst juggling an academic career I recognise the need for flexible and imaginative ways of working, enabling a diverse and productive research environment. I am very interested in promoting ED&I in this context and am currently helping to write a narrative on this subject with colleagues from other UK universities.

Research Culture

I am passionate about promoting positive research culture. I co-established the ‘Early PI network’ (ePI, https://www.bio.cam.ac.uk/biological-sciences-early-pi-network) in 2018, which I then co-chaired until 2021. This is a peer networking and mentorship initiative to aid the transition of new PIs within the School of Biological Sciences while establishing their first lab.

Research Assistant: Lucy Barlow

PhD students: Helena Crellin, Millie Race

MPhil student: Bima Assatova

Postdoctoral research assistant: Sarah Williams

The Buckley lab welcomes PhD applications and advertise projects annually via central DTP programs at UoM. There is also currently a self-funded position open in the lab: https://www.findaphd.com/phds/project/what-is-the-interrelationship-between-cell-mitosis-and-cell-polarity-during-apical-basal-polarisation-of-epithelial-organs/?p162381