Fengzhu Xiong

The symbiosis with photosynthetic dinoflagellate algae enables corals to build and sustain reef ecosystems. Individual coral polyps hold algal symbionts in their epithelial endoderm cells and lose them under environmental stress, leading to coral bleaching. How the host integrates symbionts into its body plan is not well understood. Here, using a combination of high-resolution imaging, quantitative analysis, and environmental perturbations in the sea anemone Exaiptasia diaphana (Aiptasia) and reef-building coral Pocillopora damicornis , we uncover a spatial organisation of symbionts along the aboral-oral axis of cnidarian polyps that emerges under the long-range translocation of symbionts between host cells through a fluid-filled cavity. The symbiont distribution becomes specifically enriched in the tentacle bud endoderm during Aiptasia polyp morphogenesis. This pattern can form in darkness and with algae-sized inert spheres, suggesting an innate host-intrinsic mechanism. Symbiont-occupied host cells are mechanically constrained within the endoderm and thus unable to rearrange; instead, they go through cycles of symbiont expulsion and re-uptake via the host gastric cavity, with regionally biased rates of these behaviours providing a route to enrich symbionts in the tentacles. Symbiont organisation is remodelled under increased light in adult coral polyps, with a characteristic pattern of reduced tentacle enrichment, lateral clustering and retention in the body column emerging over a timescale of days. Together, our findings reveal that the spatial organisation of symbionts is dynamically regulated in cnidarian host tissues, a capacity that may shape both the establishment of symbiosis and its resilience under environmental change.

Background: The first cell fate decisions in embryogenesis give rise to extraembryonic tissues, such as the yolk sac and placenta, which are essential for embryo survival. In mammals, extraembryonic cell types arising from these first lineage decisions show distinct DNA methylation patterning, including large partially methylated domains, gene body methylation and accumulation across developmental CpG island (CGI) promoters. Analysis of extraembryonic membranes beyond mammalian species has been limited. Using bisulphite-seq and RNA-seq, we comparatively define these epigenetic characteristics of extraembryonic tissues in both mouse and human. We then tested whether these features are conserved in the chicken extraembryonic membranes (EEMs) including the yolk sac, chorion and chorioallantoic membrane (CAM). Results: Transcriptomic analysis revealed that embryonic day (E)4 yolk sac/chorion and E14 CAM express similar marker genes to the mouse chorionic trophoblast, supporting that the chicken chorionic ectoderm shares a ‘trophoblast-like’ cell identity. We find that partially methylated domains and transcription-linked gene body methylation patterning are highly conserved between mouse and human proliferative multipotent trophoblast cells but were not evident in the chicken EEMs. However, we find that methylation of CGIs associated with developmental genes was a conserved feature of mouse, human and chicken EEMs. Conclusions: These findings support that features of the extraembryonic DNA methylation landscape have distinct underlying mechanisms. We find that only one feature is conserved across extant amniotes, suggesting distinct evolutionary pressures are associated with different features of EEM epigenetic landscape.

During neural tube patterning, a gradient of Sonic hedgehog (Shh) signaling specifies ventral progenitor fates. The cellular response to Shh is processed through a genetic regulatory network (GRN) to specify distinct fate decisions. This process integrates Shh response level, duration, and other inputs and is affected by noise in signaling and cell position. How reliably the Shh response profile predicts the fate choice of a single cell remains unclear. Here, we use live imaging to track neural progenitors in developing zebrafish and quantify both Shh and fate reporters in single cells over time. We found that there is significant heterogeneity between Shh response and fate choice in single cells. We quantitatively modeled reporter intensities to obtain single-cell response levels over time and systematically evaluated their correlation with cell fate specification. Motor neuron progenitors (pMNs) exhibit a high degree of variability in their Shh responses, which is particularly prominent in the posterior neural tube where the Shh response dynamics are similar to those of the more ventrally fated lateral floor plate cells (LFPs). Our results highlight the precision limit of morphogen-interpretation GRNs in small and dynamic target cell fields.

During neural tube patterning, a gradient of Sonic hedgehog (Shh) signaling specifies ventral progenitor fates. The cellular response to Shh is processed through a genetic regulatory network (GRN) to specify distinct fate decisions. This process integrates Shh response level, duration and other inputs and is affected by noise in signaling and cell position. How reliably Shh response profile predicts the fate choice of a single cell is unclear. Here we use live imaging to track neural progenitors in developing zebrafish and quantify both Shh and fate reporters in single cells over time. We found that there is significant heterogeneity between Shh response and fate choice in single cells. We quantitatively modeled reporter intensities to obtain single cell response levels over time and systematically evaluated their correlation with cell fate specification. Motor neuron progenitors (pMN) exhibit a high degree of variability in their Shh responses, which is particularly prominent in the posterior neural tube where the Shh response dynamics are similar to those of the more ventrally fated lateral floor plate cells (LFPs). Our results highlight the precision limit of morphogen-interpretation GRNs in small and dynamic target cell fields.

During development, spatial-temporally patterned tissue-level stresses and mechanical properties create diverse tissue shapes. To understand the mechanics of small-scale embryonic tissues, precisely controlled sensors and actuators are needed. Previously, we reported a control-based approach named tissue force microscopy (TiFM1.0), which combines dynamic positioning and imaging of an inserted cantilever probe to directly measure and impose forces in early avian embryos. Here, we present an upgraded system (TiFM2.0) that uses interferometer positioning to minimise probe holder footprint, enhancing accessibility and imaging signal. This new design enables a double-probe configuration for bidirectional stretching, compression and stress propagation experiments. As proof-of-concept, we showcase a variety of examples of TiFM2.0 applications in chicken and zebrafish embryos, including the characterisation of mechanical heterogeneities important for the morphogenesis of the chicken posterior body axis. We also present simplified designs and protocols for the replication of TiFM systems with minimal custom engineering for developmental biology labs.

Tissue spreading (epiboly) couples gastrulation to shape the initial body plan of early vertebrate embryos. How these two large-scale collective cell movements cooperate remains unclear. Here, we examine the cell mechanics and tissue dynamics underlying epiboly of the chicken blastoderm. We found that cells at the blastoderm edge undergo a wetting-like process to spread on the vitelline membrane through stiffness sensing and cytoskeleton remodelling. This interaction is robust to edge cell loss and cooperates with cell-proliferation-based blastoderm growth to drive epiboly. Surprisingly, epiboly in turn remodels the extracellular matrix (ECM) to establish a basal lamina and maintain cell-cell connections. Impairing either edge cell wetting or the ECM causes tissue thickening and buckling across the blastoderm, disrupting gastrulation movements. We conclude that epiboly facilitates gastrulation by organizing an ECM that maintains a thin blastoderm. These findings suggest a general logic of mechanical coupling between distinctly controlled tissue movements during early development.

The nasal cavities of mammals contain the maxilloturbinate bones, which are involved in reducing heat and water losses. The maxilloturbinates of Arctic seals develop into particularly elaborate labyrinthine patterns, which are well adapted to retain heat and moisture from exhaled gas. These structures develop prenatally and continue to grow postnatally. The developmental mechanism of labyrinthine patterning is unknown. Here we report a model of maxilloturbinate pattern formation in prenatal and juvenile seals based on a simple algorithmic description and three key parameters: target turbinate porosity, characteristic ossification time scale, and typical gestation time scale. Under a small set of geometrical and physical rules, our model reproduces key features of the patterns observed in the turbinate structure of three seal species. To validate our model, we measure complexity, hydraulic diameter, backbone fractal dimension, and Horton-Strahler statistics for a rigorous quantitative comparison with actual tomograms of grey and harp seal skull specimens. Our model closely replicates the structural development of seal turbinates in these respects. Labyrinthine maxilloturbinate development may depend on the ability for neighbouring bone branches to detect and avoid each other, potentially through the mechanosensing of shear stresses from amniotic fluid and air flow.

SUMMARY: During development, spatial-temporally patterned tissue-level stresses and mechanical properties create diverse tissue shapes. To understand the mechanics of small-scale embryonic tissues, precisely controlled sensors and actuators are needed. Previously, we reported a control-based approach named tissue force microscopy (TiFM1.0), which combines dynamic positioning and imaging of an inserted cantilever probe to directly measure and impose forces in early avian embryos. Here we present an upgraded system (TiFM2.0) that utilises interferometer positioning to minimise probe holder footprint, enhancing accessibility and imaging signal. This new design enables a double-probe configuration for bidirectional stretching, compression and stress propagation experiments. As proof-of-concept, we showcase a variety of examples of TiFM2.0 applications in chicken and zebrafish embryos, including the characterization of mechanical heterogeneities important for the morphogenesis of the chicken posterior body axis. We also present simplified designs and protocols for the replication of TiFM systems with minimal custom engineering for developmental biology labs.