Importance of Ca2+ in gastric epithelial restitution—new views revealed by real-time in vivo measurements

Aihara, Eitaro; Montrose, Marshall H.

doi:10.1016/j.coph.2014.07.012

Cited by 14 publications

(7 citation statements)

References 63 publications

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“…; Aihara et al . ; Aihara & Montrose, ). We have recently demonstrated that the gastric organoid system is comparable to native tissue in vivo with respect to demonstrating the shedding of dead cells into the gastric lumen with an epithelial repair time course of ∼10 min (Aihara et al .…”

Section: Discussionmentioning

confidence: 99%

Trefoil factor 2 activation of CXCR4 requires calcium mobilization to drive epithelial repair in gastric organoids

Engevik

Hanyu

Matthis

et al. 2019

The Journal of Physiology

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Key pointsr Determining the signalling cascade of epithelial repair, using murine gastric organoids, allows definition of regulatory processes intrinsic to epithelial cells, at the same time as validating and dissecting the signalling cascade with more precision than is possible in vivo r Following single cell damage, intracellular calcium selectively increases within cells adjacent to the damage site and is essential for promoting repair.r Trefoil factor 2 (TFF2) acts via chemokine C-X-C receptor 4 and epidermal growth factor receptor signalling, including extracellular signal-regulated kinase activation, to drive calcium mobilization and promote gastric repair.r Sodium hydrogen exchanger 2, although essential for repair, acts downstream of TFF2 and calcium mobilization.Abstract The gastric mucosa of the stomach is continually exposed to environmental and physiological stress factors that can cause local epithelial damage. Although much is known about the complex nature of gastric wound repair, the stepwise process that characterizes epithelial restitution remains poorly defined. The present study aimed to determine the effectors that drive gastric epithelial repair using a reductionist culture model. To determine the role of trefoil factor 2 (TFF2) and intracellular calcium (Ca 2+ ) mobilization in gastric restitution, gastric organoids were derived from TFF2 knockout (KO) mice and yellow Cameleon-Nano15 (fluorescent calcium reporter) transgenic mice, respectively. Inhibitors and recombinant protein were used to determine the upstream and downstream effectors of gastric restitution following photodamage (PD) to single cells within the gastric organoids. Single cell PD resulted in parallel events of dead cell exfoliation and migration of intact neighbouring cells to restore a continuous epithelium in the damage site. Under normal conditions following PD, Ca 2+ levels increased Kristen Engevik is a PhD candidate at the University of Cincinnati College of Medicine. Kristen's research focuses on investigating the mechanism behind gastric restitution. Her expertise lies in culturing three-dimensional gastric organoids and the use of two-photon light microscopy to induce photodamage with respect to examining the pathways involved in gastric repair. K. A. Engevik and othersJ Physiol 597.10 within neighbour migrating cells, peaking at ß1 min, suggesting localized Ca 2+ mobilization at the site of cell protrusion/migration. TFF2 KO organoids exhibit delayed repair; however, this delay can be rescued by the addition of exogenous TFF2. Inhibition of epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase (ERK)1/2 or a TFF2 receptor, chemokine C-X-C receptor 4 (CXCR4), resulted in significant delay and dampened Ca 2+ mobilization. Inhibition of sodium hydrogen exchanger 2 (NHE2) caused significant delay but did not affect Ca 2+ mobilization. A similar delay was observed in NHE2 KO organoids. In TFF2 KO gastric organoids, the addition of exogenous TFF2 in the presence of EGFR or CXCR4 inhibition was u...

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Section: Discussionmentioning

confidence: 99%

Trefoil factor 2 activation of CXCR4 requires calcium mobilization to drive epithelial repair in gastric organoids

Engevik

Hanyu

Matthis

et al. 2019

The Journal of Physiology

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“…Our work points to the critical role of voltage‐gated and SOCE Ca 2+ channels (extracellular) and PLC, IP 3 (intracellular) pathways to promoting repair. We have previously shown in vivo the importance of localized changes in both extracellular and intracellular Ca 2+ during gastric epithelial repair (Aihara et al, 2013; Aihara & Montrose, 2014) in the first study to demonstrate Ca 2+ dynamics occurring in native tissue during gastric restitution with a genetically encoded Ca 2+ indicator. Due to tissue motion in a breathing animal, it is technically challenging to examine Ca 2+ dynamics in individual cells during the response to single cell damage (unpublished observations) .…”

Section: Discussionmentioning

confidence: 99%

Multiple calcium sources are required for intracellular calcium mobilization during gastric organoid epithelial repair

et al. 2020

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Calcium (Ca2+) is a known accelerator for gastric wound repair. We have demonstrated in vivo and in vitro that intracellular Ca2+ increases in the gastric epithelial cells directly adjacent to a damaged cell, and that this Ca2+ rise is essential for the cellular migration that rapidly repairs the epithelium (restitution). While intracellular Ca2+ has been shown to be an important signaling factor during epithelial restitution, the source from which this intracellular Ca2+ originates remains unclear. Using gastric organoids derived from mice transgenic for a genetically encoded Ca2+ indicator, we sought to investigate the potential sources of intracellular Ca2+ mobilization. During confocal imaging, photodamage (PD) was induced to 1–2 gastric organoid epithelial cells and epithelial restitution measured simultaneously with changes in intracellular Ca2+ (measured as FRET/CFP ratio in migrating cells adjacent to the damaged area). Inhibition of voltage‐gated Ca2+ channels (verapamil, 10 µM) or store‐operated calcium entry (YM58483, 20 µM) resulted in delayed repair and dampened intracellular Ca2+ response. Furthermore, inhibition of phospholipase C (U73122, 10 µM) or inositol trisphosphate receptor (2‐APB, 50 µM) likewise resulted in delayed repair and dampened Ca2+ response. Results suggest both extracellular and intracellular Ca2+ sources are essential for supplying the Ca2+ mobilization that stimulates repair.

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“…Early epithelial restitution is a primary repair modality in the gastrointestinal tract, and it rapidly reseals superficial wounds by migrating visible remaining epithelial cells from areas adjacent to the injured surface to cover the wounded area, followed by proliferation and differentiation (4,5). This early rapid mucosal reepithelialization after mucosal injury is a complex process that includes the flattening, spreading, migrating, and repolarizing of differentiated columnar epithelial cells but is independent of cell proliferation (2,6). Defective regulation of early mucosal restitution underlies various critical pathological states such as massive mucosal injury and hemorrhage, delayed wound healing, disruption of epithelial integrity, and epithelial barrier dysfunction (2, 7).…”

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confidence: 99%

HuR Enhances Early Restitution of the Intestinal Epithelium by Increasing Cdc42 Translation

Liu

Zhuang

Xiao

et al. 2017

Molecular and Cellular Biology

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The mammalian intestinal mucosa exhibits a spectrum of responses after acute injury and repairs itself rapidly to restore the epithelial integrity. The RNAbinding protein HuR regulates the stability and translation of target mRNAs and is involved in many aspects of gut epithelium homeostasis, but its exact role in the regulation of mucosal repair after injury remains unknown. We show here that HuR is essential for early intestinal epithelial restitution by increasing the expression of cell division control protein 42 (Cdc42) at the posttranscriptional level. HuR bound to the Cdc42 mRNA via its 3= untranslated region, and this association specifically enhanced Cdc42 translation without an effect on the Cdc42 mRNA level. Intestinal epithelium-specific HuR knockout not only decreased Cdc42 levels in mucosal tissues, but it also inhibited repair of damaged mucosa induced by mesenteric ischemia/reperfusion in the small intestine and by dextran sulfate sodium in the colon. Furthermore, Cdc42 silencing prevented HuR-mediated stimulation of cell migration over the wounded area by altering the subcellular distribution of F-actin. These results indicate that HuR promotes early intestinal mucosal repair after injury by increasing Cdc42 translation and demonstrate the importance of HuR deficiency in the pathogenesis of delayed mucosal healing in certain pathological conditions.

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Importance of Ca2+ in gastric epithelial restitution—new views revealed by real-time in vivo measurements

Cited by 14 publications

References 63 publications

Trefoil factor 2 activation of CXCR4 requires calcium mobilization to drive epithelial repair in gastric organoids

Trefoil factor 2 activation of CXCR4 requires calcium mobilization to drive epithelial repair in gastric organoids

Multiple calcium sources are required for intracellular calcium mobilization during gastric organoid epithelial repair

HuR Enhances Early Restitution of the Intestinal Epithelium by Increasing Cdc42 Translation

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