Sustained Ca2+ mobilizations: A quantitative approach to predict their importance in cell-cell communication and wound healing

Lee, YoonJoo; Kim, Min Tae; Rhodes, Garrett; Sack, Kelsey; Son, Sung Jun; Rich, Celeste B.; Kolachalama, Vijaya B.; Gabel, Christopher V.; Trinkaus‐Randall, Vickery

doi:10.1371/journal.pone.0213422

Cited by 15 publications

(30 citation statements)

References 33 publications

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“…Reduced calcium concentrations inversely raise renin secretion levels via regulation of adenylyl cyclase and cyclic adenosine monophosphate (cAMP). 45 We hypothesized that autonomous renin secretion might be mechanistically regulated by altered intracellular calcium homeostasis because (i) Panx1 has been suggested to alter intracellular calcium levels, [46][47][48] (ii) we showed in Figure 6f an enhanced renin secretory response to EGTA, and (iii) because decreased intracellular calcium enhances renin secretion. 5,49,50 To test if intracellular calcium levels rely on extracellular ATP, we measured Fluo-4-loaded As4.1 cells in the presence of the ectoenzyme apyrase (Figure 7a).…”

Section: Resultsmentioning

confidence: 93%

Pannexin 1 channels in renin-expressing cells influence renin secretion and blood pressure homeostasis

DeLalio

Masati

Mendu

et al. 2020

Kidney International

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Kidney function and blood pressure homeostasis are regulated by purinergic signaling mechanisms. These autocrine/paracrine signaling pathways are initiated by the release of cellular ATP, which influences kidney hemodynamics and steady-state renin secretion from juxtaglomerular cells. However, the mechanism responsible for ATP release that supports tonic inputs to juxtaglomerular cells and regulates renin secretion remains unclear. Pannexin 1 (Panx1) channels localize to both afferent arterioles and juxtaglomerular cells and provide a transmembrane conduit for ATP release and ion permeability in the kidney and the vasculature. We hypothesized that Panx1 channels in renin-expressing cells regulate renin secretion in vivo. Using a renin cell-specific Panx1 knockout model, we found that male Panx1 deficient mice exhibiting a heightened activation of the reninangiotensin-aldosterone system have markedly increased plasma renin and aldosterone concentrations, and elevated mean arterial pressure with altered peripheral hemodynamics. Following ovariectomy, female mice mirrored the male phenotype. Furthermore, constitutive Panx1 channel activity was observed in As4.1 reninsecreting cells, whereby Panx1 knockdown reduced extracellular ATP accumulation, lowered basal intracellular calcium concentrations and recapitulated a hyper-secretory renin phenotype. Moreover, in response to stress stimuli that lower blood pressure, Panx1-deficient mice exhibited aberrant "renin recruitment" as evidenced by reactivation of renin expression in pre-glomerular arteriolar smooth muscle cells. Thus, renin-cell Panx1 channels suppress renin secretion and influence adaptive renin responses when blood pressure homeostasis is threatened.

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Section: Resultsmentioning

confidence: 93%

Pannexin 1 channels in renin-expressing cells influence renin secretion and blood pressure homeostasis

DeLalio

Masati

Mendu

et al. 2020

Kidney International

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“…Rapid release and rapid restoration of Ca 2+ from the intracellular stores allows fast reaction to repeated stimulus. While the utilized set-up does not allow to examine the dynamic propagation of the Ca 2+ wave between the cells, the cells treated with an agonist all react in a similar manner as cells located directly next to a wound [ 59 ]. In experiments of Lee et al 2019, the cultured corneal limbal cells located to this so-called leading edge of the wound were also shown to exhibit sustained Ca 2+ oscillations, which we also observed in our samples.…”

Section: Discussionmentioning

confidence: 99%

Corneal epithelial differentiation of human pluripotent stem cells generates ABCB5+ and ∆Np63α+ cells with limbal cell characteristics and high wound healing capacity

et al. 2021

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Background Differentiation of functional limbal stem cells (LSCs) from human pluripotent stem cells (hPSCs) is an important objective which can provide novel treatment solutions for patients suffering from limbal stem cell deficiency (LSCD). Yet, further characterization is needed to better evaluate their immunogenicity and regenerative potential before clinical applications. Methods Human PSCs were differentiated towards corneal fate and cryopreserved using a clinically applicable protocol. Resulting hPSC-LSC populations were examined at days 10–11 and 24–25 during differentiation as well as at passage 1 post-thaw. Expression of cornea-associated markers including PAX6, ABCG2, ∆Np63α, CK15, CK14, CK12 and ABCB5 as well as human leukocyte antigens (HLAs) was analyzed using immunofluorescence and flow cytometry. Wound healing properties of the post-thaw hPSC-LSCs were assessed via calcium imaging and scratch assay. Human and porcine tissue-derived cultured LSCs were used as controls for marker expression analysis and scratch assays at passage 1. Results The day 24–25 and post-thaw hPSC-LSCs displayed a similar marker profile with the tissue-derived LSCs, showing abundant expression of PAX6, ∆Np63α, CK15, CK14 and ABCB5 and low expression of ABCG2. In contrast, day 10–11 hPSC-LSCs had lower expression of ABCB5 and ∆Np63α, but high expression of ABCG2. A small portion of the day 10–11 cells coexpressed ABCG2 and ABCB5. The expression of class I HLAs increased during hPSC-LSCs differentiation and was uniform in post-thaw hPSC-LSCs, however the intensity was lower in comparison to tissue-derived LSCs. The calcium imaging revealed that the post-thaw hPSC-LSCs generated a robust response towards epithelial wound healing signaling mediator ATP. Further, scratch assay revealed that post-thaw hPSC-LSCs had higher wound healing capacity in comparison to tissue-derived LSCs. Conclusions Clinically relevant LSC-like cells can be efficiently differentiated from hPSCs. The post-thaw hPSC-LSCs possess functional potency in calcium responses towards injury associated signals and in wound closure. The developmental trajectory observed during hPSC-LSC differentiation, giving rise to ABCG2+ population and further to ABCB5+ and ∆Np63α+ cells with limbal characteristics, indicates hPSC-derived cells can be utilized as a valuable cell source for the treatment of patients afflicted corneal blindness due to LSCD. Graphical Abstract

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“…Specifically, intercellular calcium levels are synchronized via gap junction-mediated cell-cell communication 14,21 . Such synchronized calcium signaling is instrumental in modulating reepithelialization, angiogenesis, and extracellular matrix remodeling, which are essential processes in wound repair [22][23][24][25][26] . Thus, we developed an integrated experimental-computational platform to quantitatively evaluate the roles that single cells take during the emergence of multicellular synchronization.…”

Section: Introductionmentioning

confidence: 99%

Emergence of synchronized multicellular mechanosensing from spatiotemporal integration of heterogeneous single-cell information transfer

Zamir

Chase

et al. 2020

Preprint

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We quantitatively characterize how noisy and heterogeneous behaviors of individual cells are integrated across a population toward multicellular synchronization by studying the calcium dynamics in mechanically stimulated monolayers of endothelial cells. We used information-theory to quantify the asymmetric information-transfer between pairs of cells and define quantitative measures of how single cells receive or transmit information in the multicellular network. We find that cells take different roles in intercellular information-transfer and that this heterogeneity is associated with synchronization. Cells tended to maintain their roles between consecutive cycles of mechanical stimuli and reinforced them over time, suggesting the existence of a cellular "memory" in intercellular information transfer. Interestingly, we identified a subpopulation of cells characterized by higher probability of both receiving and transmitting information. These "communication hub" roles were stable - once a cell switched to a "communication hub" role it was less probable to switch to other roles. This stableness property of the cells led to gradual enrichment of communication hubs that was associated with the establishment of synchronization. Our analysis demonstrated that multicellular synchronization was established by effective information spread from the (local) single cell to the (global) group scale in the multicellular network. Altogether, we suggest that multicellular synchronization is driven by single cell communication properties, including heterogeneity, functional memory and information flow.

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Sustained Ca2+ mobilizations: A quantitative approach to predict their importance in cell-cell communication and wound healing

Cited by 15 publications

References 33 publications

Pannexin 1 channels in renin-expressing cells influence renin secretion and blood pressure homeostasis

Pannexin 1 channels in renin-expressing cells influence renin secretion and blood pressure homeostasis

Corneal epithelial differentiation of human pluripotent stem cells generates ABCB5+ and ∆Np63α+ cells with limbal cell characteristics and high wound healing capacity

Emergence of synchronized multicellular mechanosensing from spatiotemporal integration of heterogeneous single-cell information transfer

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