ΔNp63 is upregulated during salivary gland regeneration following duct ligation and irradiation in mice

Ikai, Kazuki; Sakai, Manabu; Minagi, Hitomi Ono; Gojo, Nao; Sakai, Takayoshi

doi:10.1002/1873-3468.13896

Cited by 12 publications

(11 citation statements)

References 35 publications

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“…The irradiated gland, however, does undergo some regeneration, with Sox2 cells replenishing the acinar compartment of the sublingual gland (Emmersen et al, 2018). The two methods share some similarities, for example, p63 is upregulated after both techniques (Ikai et al, 2020). It would therefore be interesting to study whether the changes in Fgf10 and Sox9 expression observed here after gland ligation also occur after irradiation.…”

Section: Discussionmentioning

confidence: 99%

Comparing development and regeneration in the submandibular gland highlights distinct mechanisms

et al. 2021

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Section: Discussionmentioning

confidence: 99%

Comparing development and regeneration in the submandibular gland highlights distinct mechanisms

et al. 2021

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“…The transcription factor p63, a component of the p53 family, plays important roles in the development, homeostasis, and regeneration of epithelial tissues [34,44]. p63 is upregulated in the epithelium of CRS and nasal polyps [46,47]. It negatively regulates the epithelial tight junctional barrier of the nasal epithelium [47].…”

Section: Discussionmentioning

confidence: 99%

Effects of HMGB1 on Tricellular Tight Junctions via TGF-β Signaling in Human Nasal Epithelial Cells

Ohwada

Konno

Kohno

et al. 2021

IJMS

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The airway epithelium of the human nasal mucosa acts as a physical barrier that protects against inhaled substances and pathogens via bicellular and tricellular tight junctions (bTJs and tTJs) including claudins, angulin-1/LSR and tricellulin. High mobility group box-1 (HMGB1) increased by TGF-β1 is involved in the induction of nasal inflammation and injury in patients with allergic rhinitis, chronic rhinosinusitis, and eosinophilic chronic rhinosinusitis. However, the detailed mechanisms by which this occurs remain unknown. In the present study, to investigate how HMGB1 affects the barrier of normal human nasal epithelial cells, 2D and 2.5D Matrigel culture of primary cultured human nasal epithelial cells were pretreated with TGF-β type I receptor kinase inhibitor EW-7197 before treatment with HMGB1. Knockdown of angulin-1/LSR downregulated the epithelial barrier. Treatment with EW-7197 decreased angulin-1/LSR and concentrated the expression at tTJs from bTJs and increased the epithelial barrier. Treatment with a binder to angulin-1/LSR angubindin-1 decreased angulin-1/LSR and the epithelial barrier. Treatment with HMGB1 decreased angulin-1/LSR and the epithelial barrier. In 2.5D Matrigel culture, treatment with HMGB1 induced permeability of FITC-dextran (FD-4) into the lumen. Pretreatment with EW-7197 prevented the effects of HMGB1. HMGB1 disrupted the angulin-1/LSR-dependent epithelial permeability barriers of HNECs via TGF-β signaling in HNECs.

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“…Since then, the approach has been used for organ culture of various tissues. In 2008, we reported the first detailed protocol for a 3D submandibular salivary gland organ cell culture (T. Sakai & Onodera, 2008), which later became the standard method for organ culture of submandibular salivary glands (Ikai, Sakai, Minagi, Gojo, & Sakai, 2020; Knosp et al., 2012; M. Sakai et al., 2019).…”

Section: Introductionmentioning

confidence: 99%

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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ΔNp63 is upregulated during salivary gland regeneration following duct ligation and irradiation in mice

Cited by 12 publications

References 35 publications

Comparing development and regeneration in the submandibular gland highlights distinct mechanisms

Comparing development and regeneration in the submandibular gland highlights distinct mechanisms

Effects of HMGB1 on Tricellular Tight Junctions via TGF-β Signaling in Human Nasal Epithelial Cells

Establishment of a Mouse Submandibular Salivary Gland Organ Culture

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