Biology subjects

Manning, S. A.

Publications and source records attributed to Manning, S. A..

5 recordsLinked to original sources

bioRxiv2026

Hippo pathway perturbation disrupts cell fate control in the Drosophila eye

During animal development, cells acquire specialised fates in a precise spatiotemporal order, which is essential for producing tissues that function appropriately. Cell fate specification is governed by multiple signalling pathways, as well as mechanical forces, which impact cellular transcription. Two such signalling pathways are the Hippo pathway and EGFR pathway, which both control organ growth and the fate of certain cell types in multiple species. Here, we show that Hippo signalling is essential for the maintenance of the cone and primary pigment cell fates in the developing Drosophila eye. When Hippo signalling is compromised, its nuclear effectors Yorkie and Scalloped drive increased expression of the EGFR pathway transcription repressor Yan, which antagonises the cone and primary pigment cell fates. Thus, in addition to its role as a growth suppressor, Hippo signalling promotes the fate of multiple eye cells by maintaining their responsiveness to inductive cues from the EGFR pathway. AUTHOR SUMMARYAs multicellular organisms grow and develop from a zygote, individual cells become increasingly specialised. Cell fate is specified and maintained by the coordinate action of different signalling pathways, whose activity must be tightly controlled in a spatiotemporal fashion. If this fails, cell fate can be disturbed, which can cause developmental abnormalities, and diseases such as cancers. Here, we describe a new function for the Hippo signalling pathway, which was originally discovered as a regulator of Drosophila tissue growth and subsequently linked to the genesis of multiple human cancers. Hippo signalling is essential for maintaining the fate of two key cell types in the Drosophila eye, primary pigment cells and cone cells. Without Hippo signalling these cells cannot properly respond to signals from another key signalling network, the EGFR pathway. Our discoveries add to a growing literature where the Hippo growth control pathway is repurposed to control cell fate in tissues that have ceased growth.

developmental biology↗

bioRxiv2026

Observing concurrent subcellular dynamics in large living tissues

An outstanding question in eukaryotic biology is the mechanistic connection between events occurring at (sub)cellular levels (time scales of milliseconds to minutes) to those at the tissue levels (tens of minutes to months). Deciphering such mechanisms requires imaging approaches capable of simultaneously achieving high spatial and temporal resolutions for large samples over long periods of time. Here, we demonstrate Airy beam-based light sheet microscopy of organelles in tens to hundreds of cells in a few hundred micrometre-wide tissue environments. We achieve a typical resolution of 320 nm over 266 x 266 x 100 m3 volumes at a temporal rate of 0.05 Hz, typically with generally used fluorophores such as Green Fluorescent Protein, over extended periods of time that allow tracking of organelle and protein dynamics. We validated our approach across different length and time scales by imaging mitochondria and endosome dynamics in very large fields of view in zebrafish tissue, molecular assemblies of myosin as gastrulation proceeds in Drosophila embryos, 3D mitochondrial streaming in mouse oocytes, pressure-driven motility and protrusions in amoebae, mitochondrial dynamics in cancer spheroids, 5 -colour fast imaging in iBlastoids, and endosomal dynamics in single cells. Through these model systems, we demonstrate the versatility of Airy beam light sheet microscopy to image large tissues at unprecedented high resolution; to capture dynamics in photosensitive, delicate samples; and to screen 3D samples. We anticipate that our Airy beam-based approach will represent a pivotal advance in cellular biology--especially developmental biology--as it provides, for the first time, true subcellular resolution over large imaging volumes with high temporal resolution.

developmental biology↗

bioRxiv2026

The mechanosensitive protein Zyxin influences Hippo signalling and tissue growth via adherens junctions and basal spot junctions in Drosophila

Physical forces like tension and compression have a central role in the growth and morphogenesis of tissues. Cell-cell junctions and the actin cytoskeleton are key mediators of physical forces and influence activity of signalling networks like the Hippo pathway. Here, we show that the mechanosensitive proteins Zyxin and Ajuba work in concert to limit Hippo signalling and promote Drosophila epithelial tissue growth. Zyxin recruits both Ajuba and the central Hippo pathway kinase Warts to adherens junctions and basal spot junctions, and limit Warts activity by segregating it from Hippo pathway activator proteins. Zyxin also promotes the transmission of actomyosin tension to adherens junctions in growing epithelial tissues, indicating conservation of this function. Thus, we provide new insights into how Zyxin couples mechanical cues and Hippo pathway-dependent tissue growth and highlight the importance of basal spot junctions and their associated actomyosin network in epithelial tissue development.

developmental biology↗

bioRxiv2025

Hippo signalling regulates the nuclear behaviour and DNA dwell times of YAP and TEAD to control transcription

Over the past two decades, genetic and proteomic screens have enabled the discovery and elucidation of the Hippo pathway as a complex signalling network that controls tissue growth and cell fate and which is of major importance for human cancers. Despite these advances, our understanding of how Hippo signalling regulates transcription is less clear. To address this, we used live microscopy approaches to study the nuclear behaviour of the major transcription effectors of the human Hippo pathway, YAP and TEADs. Our experiments revealed that TEADs are a major determinant of YAPs nuclear biophysical behaviour, whilst YAP only has a minor influence on TEAD behaviour. Acute chemical inhibition of Hippo signalling stimulated an increase in the DNA residence time of both YAP and TEAD1. Consistently, YAP and TEAD1 bound DNA for longer periods in cells with high intrinsic YAP/TEAD activity (induced trophoblast stem cells) than in cells with low intrinsic YAP/TEAD activity (induced pluripotent stem cells). TEAD1 bound the genome on a broad range of timescales, and this is extended substantially in nuclear condensates. Finally, single molecule tracking experiments revealed that a fusion protein encoded by a cancer-associated YAP allele exhibits substantially different nuclear biophysical behaviour than either YAP or TEAD1. These live microscopy experiments reveal that Hippo signalling regulates transcription at least in part by influencing the DNA dwell times of both YAP and TEAD.

cell biology↗

bioRxiv2023

Basal spot junctions of epithelial tissues respond to morphogenetic forces and regulate Hippo signalling

Organ size is controlled by numerous factors including mechanical forces, which are mediated in part by the Hippo pathway. In growing Drosophila melanogaster epithelial tissues, cytoskeletal tension influences Hippo signalling by modulating the subcellular localisation of key pathway proteins in different apical domains, namely adherens junctions, the sub-apical region and the medial apical cortex. Here, using both electron and light microscopy, we have discovered the existence of basal spot junctions in D. melanogaster epithelial tissues, and that they respond to morphogenetic forces and also influence Hippo signalling. Like adherens junctions, the Warts kinase is recruited to basal spot junctions via Ajuba and E-cadherin, which prevent Warts activation by segregating it from upstream Hippo pathway proteins. Basal spot junctions are prominent when tissues undergo morphogenesis and are highly sensitive to fluctuations in cytoskeletal tension. Basal spot junctions are distinct from focal adhesions, but the latter profoundly influences the abundance of spot junctions by modulating the basal-medial actomyosin network and tension experienced by spot junctions. Thus, basal spot junctions potentially couple morphogenetic forces to Hippo pathway activity and organ growth.

cell biology↗