Tissue mechanics drives regeneration of a mucociliated epidermis on the surface of Xenopus embryonic aggregates

Kim, Hye Young; Jackson, Timothy R.; Stuckenholz, Carsten; Davidson, Lance A.

doi:10.1038/s41467-020-14385-y

Cited by 25 publications

(22 citation statements)

References 55 publications

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“…The YAP transcription factor, a central component of the Hippo signaling pathway, responds to cell surface tension and mechanical forces, regulates epithelial size during airway development and has been linked to both actin cytoskeletal rearrangements and ciliogenesis [113,114]. YAP is expressed in mouse airway MCCs, several YAP interactors have been localized to BBs, and in frogs, YAP nuclear translocation and increased tissue stiffness occur during regeneration of the multiciliated epithelium [115,116]. However, YAP is not required for MCC generation per se and it remains unclear if YAP plays an important role in dictating MCC size or centriole numbers [117].…”

Section: Late Transcriptional Roles Polarization and Cilia Assemblymentioning

confidence: 99%

Transcriptional regulation of multiciliated cell differentiation

Lewis

Stracker

2021

Seminars in Cell & Developmental Biology

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GEMC1 and MCIDAS control multiciliated cell differentiation in a stepwise manner.• p73 plays a major role in multiciliogenesis.• Multiciliated cells activate a cell cycle program to regulate centriole amplification.• PLK4, mother centrioles and deuterosomes are not strictly required for multiciliogenesis.• Centriole numbers scale to cell surface area. Multiciliated cells in vertebratesMulticiliated cells (MCCs) are specialized epithelial cells that project multiple motile cilia required for respiratory, reproductive, renal and brain functions in many vertebrates [1]. In humans, MCCs are present in the ependyma and choroid plexus of the brain to direct the flow of cerebrospinal fluid, the airways to clear mucus and pathogens, and in the efferent ducts and oviducts for spermatozoa and egg transport, respectively. Depending on the tissue, dozens to hundreds of motile cilia are generated per MCC that can beat in a coordinated, directional manner or generate turbulence through whip-likeThe process of MCC differentiation, or multiciliogenesis, requires the activation of a unique transcriptional program that specifies cell fate and allows the massive amplification of centrioles; barrel-shaped, microtubule based organelles that dock at the cell surface with other factors to provide a basal body (BB) required to support the generation of the ciliary axoneme [3]. As several recent reviews have covered different aspects of multiciliogenesis in great detail [1,[3][4][5][6][7][8][9][10], here we will focus on providing an overview of the transcriptional regulation of multiciliogenesis and how it connects to key cellular processes. In addition, we will highlight recent advances in our understanding of other cell biological aspects of MCC development and the consequences of their dysfunction. MCC specification: Notch and the Geminin family proteinsThe inhibition of Notch signaling has emerged as a consistent early event in MCC differentiation from the study of frog skin, zebrafish pronephros and murine ependymal, fallopian tube (oviduct) and airway epithelia (Figure 1) [11][12][13][14][15][16][17][18]. The precise details of Notch regulation remain unclear in all cases but the Mir-34/449 family of miRNAs has been implicated in Notch inhibition in frogs, zebrafish and mice and this miRNA family plays redundant roles in MCC formation in several tissues, including the brain, airway and male germline [2,11,13,[19][20][21][22][23]. In the murine airway, progenitor cells give rise to secretory (Clara) or MCC lineages in a Notch dependent manner[24]. Genetic or pharmacological inhibition of Notch causes the trans-differentiation of secretory cells into MCCs, demonstrating a central role for Notch inhibition in initiating the MCC differentiation program in the airway[15]. The full effects of Notch inhibition on transcription have not been clearly elucidated at early steps of MCC differentiation, but fate decisions following Notch inhibition are controlled by the interplay between the Geminin family proteins; Geminin (encoded by GMNN), ...

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Section: Late Transcriptional Roles Polarization and Cilia Assemblymentioning

confidence: 99%

Transcriptional regulation of multiciliated cell differentiation

Lewis

Stracker

2021

Seminars in Cell & Developmental Biology

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“…This standardized protocol generates a mucociliary epithelial organoid from multipotent progenitors isolated from the early gastrula stage X. laevis embryos within 24 h of cultivation 14 Figure 1C).…”

Section: Representative Resultsmentioning

confidence: 99%

“…and the development of mucociliary epithelia15 , 17 , 22 that utilize the intact ectoderm, the deep ectoderm-derived organoids presented in this protocol offer a distinct opportunity to monitor the tissue mechanics-driven regeneration phases of the surface epithelium14 . At around 2 hpa, the newlyCopyright © 2020 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License jove.com July 2020 • 161 • e61604 • Page 9 of 11 generated ZO-1 positive epithelial cells (Figure 2) begin to appear on the apical surface of organoids and increase their population to cover the entire organoid as the tissue solidify or reduces the compliance 14 .…”

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confidence: 99%

Mucociliary Epithelial Organoids from Xenopus Embryonic Cells: Generation, Culture and High-Resolution Live Imaging

Kang

Kim

2020

JoVE

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Mucociliary epithelium provides the first line of defense by removing foreign particles through the action of mucus production and cilia-mediated clearance. Many clinically relevant defects in the mucociliary epithelium are inferred as they occur deep within the body. Here, we introduce a tractable 3D model for mucociliary epithelium generated from multipotent progenitors that were microsurgically isolated from Xenopus laevis embryos. The mucociliary epithelial organoids are covered with newly generated epithelium from deep ectoderm cells and later decorated with distinct patterned multiciliated cells, secretory cells, and mucus-producing goblet cells that are indistinguishable from the native epidermis within 24 h. The full sequences of dynamic cell transitions from mesenchymal to epithelial that emerge on the apical surface of organoids can be tracked by high-resolution live imaging. These in vitro cultured, selforganizing mucociliary epithelial organoids offer distinct advantages in studying the biology of mucociliary epithelium with high-efficiency in generation, defined culture conditions, control over number and size, and direct access for live imaging during the regeneration of the differentiated epithelium. associated with impaired production and clearance of mucus which is often found in pulmonary disorders such as chronic obstructive pulmonary disease, asthma, cystic fibrosis, bronchiectasis, and primary ciliary dyskinesia 1 , 2 , 3 , 4. A recent advance in organoid technology, for instance, the basal cell derived lung organoid called tracheosphere that recapitulates the regeneration of mucociliary epithelium arise as a promising model with therapeutic potential 1 , 5 , 6. However, its use is currently limited, in part because of lack of the defined culture conditions and low efficiency in organoid productions. Mucociliary epithelium in the human airway and frog epidermis are remarkably similar in tissue morphology, cellular composition, and its function 7 , 8 , 9 , 10 , 11 , 12. In both organisms, mucociliary epithelium provides first-line defense by secreting mucus and antimicrobial substances and clears Copyright © 2020 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License jove.com July 2020 • 161 • e61604 • Page 2 of 11 harmful particles and pathogens through the synchronized action of cilia. Here, we describe a simple protocol to generate mucociliary epithelial organoids using the multipotent progenitors of Xenopus laevis embryos 13 , 14. Previously, we reported 14 that in the absence of exogenous growth factors and the extracellular matrix, the microsurgically isolated deep cells from the early gastrula stage ectoderm spontaneously assemble into aggregates, regenerate epithelium on its surface, and mature into mucociliary epithelium by intercalating multiciliated and other accessory cells within 24 h. In addition to the rapid development, this protocol offers a distinct opportunity for directly accessing the transitions of multipotent deep ectoderm cells into epithelial gob...

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“…Injection of mRNA, morpholino or CRISPR constructs followed by Isolation of pluripotent stem cells from blastula stage is and accessible technique. At the blastula stage, the cells sitting on top of the blastocoel, called the animal cap, can be explanted, and differentiated ex vivo into many different tissue types, including kidney, neuronal and mucociliary epidermal tissues (Kim, Jackson, Stuckenholz, & Davidson, 2020; Li, Mofunanya, Harris, & Takemaru, 2008; Sater, Steinhardt, & Keller, 1993; Uochi & Asashima, 1996). Given that each of these cells contain yolk, they will stay viable in a saline solution at room temperature for many days.…”

Section: Motile Ciliamentioning

confidence: 99%

Aquatic models of human ciliary diseases

Corkins

Krneta‐Stankic

Kloc

et al. 2021

Genesis

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Cilia are microtubule‐based structures that either transmit information into the cell or move fluid outside of the cell. There are many human diseases that arise from malfunctioning cilia. Although mammalian models provide vital insights into the underlying pathology of these diseases, aquatic organisms such as Xenopus and zebrafish provide valuable tools to help screen and dissect out the underlying causes of these diseases. In this review we focus on recent studies that identify or describe different types of human ciliopathies and outline how aquatic organisms have aided our understanding of these diseases.

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Tissue mechanics drives regeneration of a mucociliated epidermis on the surface of Xenopus embryonic aggregates

Cited by 25 publications

References 55 publications

Transcriptional regulation of multiciliated cell differentiation

Transcriptional regulation of multiciliated cell differentiation

Mucociliary Epithelial Organoids from Xenopus Embryonic Cells: Generation, Culture and High-Resolution Live Imaging

Aquatic models of human ciliary diseases

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