The Wnt-Myb pathway suppresses KIT expression to control the timing of salivary proacinar differentiation and duct formation

Matsumoto, Shinji; Kurimoto, Tetsuya; Taketo, Makoto Mark; Fujii, Shinsuke; Kikuchi, Akira

doi:10.1242/dev.134486

Cited by 35 publications

(44 citation statements)

References 40 publications

(47 reference statements)

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“…Although our data do not delineate a specific mechanism for endothelial cell signaling in controlling SMG cell fate, they are consistent with data from other embryonic organs indicating that the developing vasculature supports expansion of the primitive progenitor populations, which in turn recruits developing vasculature to the emerging epithelium. A similar premature ductal differentiation was recently reported during early development of the salivary gland that is Wnt dependent (Matsumoto et al, 2016). Similar to our data demonstrating premature ductal formation and expansion of the ductal area of the gland with compromised VEGFR2/CD31 + endothelial cell function, Matsumoto et al observed that excess Wnt signaling can induce premature ductal formation and expansion of the ductal compartment of the gland.…”

Section: Discussionsupporting

confidence: 92%

“…Loss of VEGFR2/CD31 + endothelial cell function would then be expected to increase canonical Wnt signaling to the epithelium, prevent proacinar differentiation, downregulate Kit expression and accelerate ductal formation. Wnt signaling is regulated by balancing activation of Wnt receptors by ligands at many levels, and several reports indicate that the Wnt family has complex roles in SMG development (Haara et al, 2011;Musselmann et al, 2011;Patel et al, 2011;Knosp et al, 2015;Maimets et al, 2016;Matsumoto et al, 2016). Thus, VEGFR2/CD31 + endothelial cells could act in many ways to modulate mesenchymal Wnt signaling to the epithelium, and might confer this regulation in a spatially restricted manner during branching morphogenesis.…”

Section: Discussionmentioning

confidence: 99%

“…We examined the productal marker K19 together with Kit, a tyrosine kinase expressed by proacinar cells in developing salivary glands (Nelson et al, 2013;Lombaert et al, 2013;Matsumoto et al, 2016). In whole organ explant cultures, VEGFR signaling and vasculature development were manipulated using the pharmacological VEGFR2 inhibitors ZM 323881 and SU 5416.…”

Section: Vegfr Signaling and Vasculature Of Smgs Regulate Epithelial mentioning

confidence: 99%

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Endothelial cell regulation of salivary gland epithelial patterning

et al. 2017

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Perfusion-independent regulation of epithelial pattern formation by the vasculature during organ development and regeneration is of considerable interest for application in restoring organ function. During murine submandibular salivary gland development, the vasculature co-develops with the epithelium during branching morphogenesis; however, it is not known whether the vasculature has instructive effects on the epithelium. Using pharmacological inhibitors and siRNA knockdown in embryonic organ explants, we determined that VEGFR2-dependent signaling is required for salivary gland epithelial patterning. To test directly for a requirement for endothelial cells in instructive epithelial patterning, we developed a novel ex vivo cell fractionation/reconstitution assay. Immunodepletion of CD31 + endothelial cells in this assay confirmed a requirement for endothelial cells in epithelial patterning of the gland. Depletion of endothelial cells or inhibition of VEGFR2 signaling in organ explants caused an aberrant increase in cells expressing the ductal proteins K19 and K7, with a reduction in Kit + progenitor cells in the endbuds of reconstituted glands. Addition of exogenous endothelial cells to reconstituted glands restored epithelial patterning, as did supplementation with the endothelial cell-regulated mesenchymal factors IGFBP2 and IGFBP3. Our results demonstrate that endothelial cells promote expansion of Kit + progenitor cells and suppress premature ductal differentiation in early developing embryonic submandibular salivary gland buds.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Vegfr Signaling and Vasculature Of Smgs Regulate Epithelial mentioning

confidence: 99%

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Endothelial cell regulation of salivary gland epithelial patterning

et al. 2017

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“…By administering tamoxifen to pregnant mice at day 12.5 of gestation, Cre-mediated recombination was induced and β-catenin was stabilized in embryos at E17.5 (Matsumoto et al, 2016). Compared with Ctnnb1 (Ex3)fl/fl (control) embryos at E17.5, ppMLC, α18 and Factin in adherens junctions and podocalyxin in the apical cortices of lung epithelium were clearly observed in Ctnnb1 (Ex3)fl/fl ;Cre embryos as seen in control embryos at E14.5 (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…All animal experiments were carried out according to Osaka University guidelines for the care and use of experimental animals. Tamoxifen administration in mice was performed as previously described (Matsumoto et al, 2016). Mark1 mutant mice were generated using the triple CRISPR method as previously described (Sunagawa et al, 2016).…”

Section: Micementioning

confidence: 99%

Modulation of apical constriction by Wnt signaling is required for lung epithelial shape transition

Fumoto

Takigawa-Imamura

Sumiyama³

et al. 2016

Development

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In lung development, the apically constricted columnar epithelium forms numerous buds during the pseudoglandular stage. Subsequently, these epithelial cells change shape into the flat or cuboidal pneumocytes that form the air sacs during the canalicular and saccular (canalicular-saccular) stages, yet the impact of cell shape on tissue morphogenesis remains unclear. Here, we show that the expression of Wnt components is decreased in the canalicularsaccular stages, and that genetically constitutive activation of Wnt signaling impairs air sac formation by inducing apical constriction in the epithelium as seen in the pseudoglandular stage. Organ culture models also demonstrate that Wnt signaling induces apical constriction through apical actomyosin cytoskeletal organization. Mathematical modeling reveals that apical constriction induces bud formation and that loss of apical constriction is required for the formation of an air sac-like structure. We identify MAP/microtubule affinity-regulating kinase 1 (Mark1) as a downstream molecule of Wnt signaling and show that it is required for apical cytoskeletal organization and bud formation. These results suggest that Wnt signaling is required for bud formation by inducing apical constriction during the pseudoglandular stage, whereas loss of Wnt signaling is necessary for air sac formation in the canalicular-saccular stages.

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Establishment of a Mouse Submandibular Salivary Gland Organ Culture

Sakai

2022

Current Protocols

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The salivary glands produce saliva and are important in maintaining oral health. Saliva keeps the mouth moist, cleanses the oral cavity, aids digestion, and has antibacterial properties. Saliva also helps in swallowing and speech. Investigating the development of the salivary glands is thus relevant in the context of both health and disease. Various cell culture methods have been used to study salivary gland development, including culturing cells in two dimensions (2D). Under physiological conditions, cells constantly interact with other cells and the extracellular matrix, which controls complex biological functions such as cell migration and apoptosis, and can modulate gene expression. Since many of these functions are not accurately represented or reproduced in 2D culture, the results of in vitro experiments using such culture methods are often not reflected in vivo. The use of 3D cultures, such as organ cultures, has helped address this issue and has emerged as a model that better reflects the in vivo physiological environment. Here, we describe a protocol for establishing submandibular salivary gland organ culture that is more concise and simpler than previous methods and includes the separation and dissection of the salivary glands. We also describe the use of environmental stress (hypoxic stimulation) and inhibitors (U0126, LY294002, and rapamycin) to elucidate signaling pathways involved in salivary gland development. This protocol can provide researchers with a simpler and more robust method of salivary gland organ culture, enabling analysis of organ‐based signaling pathways to advance developmental biology research. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Submandibular salivary gland organ culture Basic Protocol 2: Analysis of salivary gland development in the presence of hypoxia and signaling pathway inhibitors Basic Protocol 3: Western blotting using submandibular salivary gland organ culture

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The Wnt-Myb pathway suppresses KIT expression to control the timing of salivary proacinar differentiation and duct formation

Cited by 35 publications

References 40 publications

Endothelial cell regulation of salivary gland epithelial patterning

Endothelial cell regulation of salivary gland epithelial patterning

Modulation of apical constriction by Wnt signaling is required for lung epithelial shape transition

Establishment of a Mouse Submandibular Salivary Gland Organ Culture

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