scholarly journals Three-axis classification of mouse lung mesenchymal cells reveals two populations of myofibroblasts

2021 ◽  
Author(s):  
Odemaris Narvaez del Pilar ◽  
Jichao Chen
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Single Cell ◽  
Mesenchymal Cell ◽  
Muscle Cells ◽  
Cell Types ◽  
Lineage Tracing ◽  
Mouse Lung ◽  
Airway Smooth Muscle Cells ◽  
Two Populations

The mesenchyme consists of heterogeneous cell populations that support neighboring structures and are integral to intercellular signaling. Despite such importance, mesenchymal cell types are poorly defined morphologically and molecularly, lagging behind their counterparts in the epithelial, endothelial, and immune lineages. Leveraging single-cell RNA-seq, three-dimensional imaging, and lineage tracing, we classify the mouse lung mesenchyme into three proximal-distal axes that are associated with the endothelium, epithelium, and interstitium, respectively. From proximal to distal, (1) the vascular axis includes vascular smooth muscle cells and pericytes that transition as arterioles and venules ramify into capillaries; (2) the epithelial axis includes airway smooth muscle cells and two populations of myofibroblasts: ductal myofibroblasts, surrounding alveolar ducts and marked by CDH4, HHIP, and Lgr6, which persist post-alveologenesis, and alveolar myofibroblasts, surrounding alveoli and marked by high expression of PDGFRA, which undergo developmental apoptosis; (3) the interstitial axis, residing between the epithelial and vascular trees and sharing a newly-identified marker MEOX2, includes fibroblasts in the bronchovascular bundle and the alveolar interstitium that are marked by IL33/DNER/PI16 and Wnt2, respectively. Single-cell imaging reveals distinct morphology of each mesenchymal cell population. This classification provides a conceptual and experimental framework applicable to other organs.

ATP stimulates Ca2+oscillations and contraction in airway smooth muscle cells of mouse lung slices

2002 ◽  
Vol 283 (6) ◽  
pp. L1271-L1279 ◽  
Author(s):  
Albrecht Bergner ◽  
Michael J. Sanderson
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Phospholipase C ◽  
Airway Smooth Muscle ◽  
Muscle Cells ◽  
Mouse Lung ◽  
Airway Smooth Muscle Cells ◽  
Airway Caliber ◽  
Inositol 1,4,5 Trisphosphate ◽  
Trisphosphate Receptor

In airway smooth muscle cells (SMCs) from mouse lung slices, ≥10 μM ATP induced Ca2+oscillations that were accompanied by airway contraction. After ∼1 min, the Ca2+oscillations subsided and the airway relaxed. By contrast, ≥0.5 μM adenosine 5′- O-(3-thiotriphosphate) (nonhydrolyzable) induced Ca2+oscillations in the SMCs and an associated airway contraction that persisted for >2 min. Adenosine 5′- O-(3-thiotriphosphate)-induced Ca2+oscillations occurred in the absence of external Ca2+but were abolished by the phospholipase C inhibitor U-73122 and the inositol 1,4,5-trisphosphate receptor inhibitor xestospongin. Adenosine, AMP, and α,β-methylene ATP had no effect on airway caliber, and the magnitude of the contractile response induced by a variety of nucleotides could be ranked in the following order: ATP = UTP > ADP. These results suggest that the SMC response to ATP is impaired by ATP hydrolysis and mediated via P2Y2or P2Y4receptors, activating phospholipase C to release Ca2+via the inositol 1,4,5-trisphosphate receptor. We conclude that ATP can serve as a spasmogen of airway SMCs and that Ca2+oscillations in SMCs are required to sustain airway contraction.

scholarly journals A smooth muscle-like niche facilitates lung epithelial regeneration

2019 ◽  
Author(s):  
Alena Moiseenko ◽  
Ana Ivonne Vazquez-Armendariz ◽  
Xuran Chu ◽  
Stefan Günther ◽  
Kevin Lebrigand ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Single Cell ◽  
Airway Smooth Muscle ◽  
Mesenchymal Cell ◽  
Lineage Tracing ◽  
Airway Smooth Muscle Cells ◽  
Genetic Lineage ◽  
Lung Field ◽  
Epithelial Regeneration ◽  
Cell Subpopulations

AbstractThe mammalian lung is a highly complex organ due to its branched, tree-like structure and diverse cellular composition. Recent efforts using state-of-the-art genetic lineage tracing and single-cell transcriptomics have helped reduce this complexity and delineate the ancestry and fate of various cell subpopulations during organogenesis, homeostasis and repair after injury. However, mesenchymal cell heterogeneity and function in development and disease remain a longstanding issue in the lung field. In this study, we break down smooth muscle heterogeneity into the constituent subpopulations by combiningin vivolineage tracing, single-cell RNA sequencing andin vitroorganoid cultures. We identify a repair-supportive mesenchymal cell (RSMC) population that is distinct from pre-existing airway smooth muscle cells (ASMC) and is critical for regenerating the conducting airway epithelium. Progenitors of RSMCs are intertwined with airway smooth muscle, undergo active WNT signaling, transiently acquire the expression of the smooth muscle marker ACTA2 in response to epithelial injury and are marked by PDGFRα expression. Our data simplify the cellular complexity of the peribronchiolar domain of the adult lung and represent a forward step towards unraveling the role of mesenchymal cell subpopulations in instructing epithelial behavior during repair processes.

scholarly journals Single cell RNA sequencing redefines the mesenchymal cell landscape of mouse endometrium

2020 ◽  
Author(s):  
PM Kirkwood ◽  
DA Gibson ◽  
JR Smith ◽  
JR Wilson-Kanamori ◽  
O Kelepouri ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Single Cell ◽  
Rna Sequencing ◽  
Vascular Smooth Muscle Cells ◽  
Mesenchymal Cell ◽  
Muscle Cells ◽  
Perivascular Niche ◽  
Single Cell Rna Sequencing

AbstractThe endometrium is a dynamic tissue that exhibits remarkable resilience to repeated episodes of differentiation, breakdown, regeneration and remodelling. Endometrial physiology relies on a complex interplay between the stromal and epithelial compartments with the former containing a mixture of fibroblasts, vascular and immune cells. There is evidence for rare populations of putative mesenchymal progenitor cells located in the perivascular niche of human endometrium, but the existence of an equivalent cell population in mouse is unclear.In the current study we used the Pdgfrb-BAC-eGFP transgenic reporter mouse in combination with bulk and single cell RNA sequencing (scRNAseq) to redefine the endometrial mesenchyme. Contrary to previous reports we show that CD146 is expressed in both PDGFRβ+ perivascular cells as well as CD31+ endothelial cells. Bulk RNAseq revealed cells in the perivascular niche which express high levels of Pdgfrb as well as genes previously identified in pericytes and/or vascular smooth muscle cells (Acta2, Myh11, Olfr78, Cspg4, Rgs4, Rgs5, Kcnj8, Abcc9). scRNAseq identified five subpopulations of cells including closely related pericytes/vascular smooth muscle cells and three subpopulations of fibroblasts. All three fibroblast populations were PDGFRα+/CD34+ but were distinct in their expression of Spon2/Angptl7 (fibroblast 1), Smoc2/Rgs2 (fibroblast 2) and Clec3b/Col14a1/Mmp3 (fibroblast 3), with potential functions in regulation of immune responses, response to wounding and organisation of extracellular matrix respectively.In conclusion, these data are the first to provide a single cell atlas of the mesenchymal cell landscape in mouse endometrium. By identifying novel markers for subpopulations of mesenchymal cells we can use mouse models investigate their contribution to endometrial function, compare with other tissues and apply these findings to further our understanding of human endometrium.HighlightsGFP expression in the mouse endometrium, under the control of the Pdgfrb promoter, is restricted to two cell populations based on the intensity of GFP with GFPbright cells close to the vasculatureSingle cell RNAseq identified five subpopulations of GFP+ mesenchymal cells: pericytes, vascular smooth muscle cells (vSMC) and three closely related but distinct populations of fibroblastsBioinformatics revealed that pericytes and vSMC share functions associated with the circulatory system, actin-filament process and cell adhesion, and an apparent role for pericytes in smooth muscle cell migration and response to interferonsComparisons between the fibroblast subpopulations suggest distinct roles in regulation of immune response, response to wound healing and collagen organisation.Graphical Abstract

A method for isolating adult and neonatal airway smooth muscle cells and measuring shortening velocity

1999 ◽  
Vol 86 (1) ◽  
pp. 427-435 ◽  
Author(s):  
S. P. Driska ◽  
R. E. Laudadio ◽  
M. R. Wolfson ◽  
T. H. Shaffer
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Single Cell ◽  
Airway Smooth Muscle ◽  
Smooth Muscles ◽  
Muscle Cells ◽  
Cell Length ◽  
Airway Smooth Muscle Cells ◽  
Shortening Velocity ◽  
Adult Cells

Methods are described for isolating smooth muscle cells from the tracheae of adult and neonatal sheep and measuring the single-cell shortening velocity. Isolated cells were elongated, Ca2+ tolerant, and contracted rapidly and substantially when exposed to cholinergic agonists, KCl, serotonin, or caffeine. Adult cells were longer and wider than preterm cells. Mean cell length in 1.6 mM CaCl2 was 194 ± 57 (SD) μm ( n = 66) for adult cells and 93 ± 32 μm ( n = 20) for preterm cells ( P < 0.05). Mean cell width at the widest point of the adult cells was 8.2 ± 1.8 μm ( n = 66) and 5.2 ± 1.5 μm ( n = 20) for preterm cells ( P < 0.05). Cells were loaded into a perfusion dish maintained at 35°C and exposed to agonists, and contractions were videotaped. Cell lengths were measured from 30 video frames and plotted as a function of time. Nonlinear fitting of cell length to an exponential model gave shortening velocities faster than most of those reported for airway smooth muscle tissues. For a sample of 10 adult and 10 preterm cells stimulated with 100 μM carbachol, mean (± SD) shortening velocity of the preterm cells was not different from that of the adult cells (0.64 ± 0.30 vs. 0.54 ± 0.27 s−1, respectively), but preterm cells shortened more than adult cells (68 ± 12 vs. 55 ± 11% of starting length, respectively; P < 0.05). The preparative and analytic methods described here are widely applicable to other smooth muscles and will allow contraction to be studied quantitatively at the single-cell level.

Ginger attenuates acetylcholine-induced contraction and Ca2+ signalling in murine airway smooth muscle cells

2008 ◽  
Vol 86 (5) ◽  
pp. 264-271 ◽  
Author(s):  
Muhammad N. Ghayur ◽  
Anwar H. Gilani ◽  
Luke J. Janssen
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Airway Smooth Muscle ◽  
Zingiber Officinale ◽  
Muscle Cells ◽  
Chronic Illnesses ◽  
Mouse Lung ◽  
Airway Smooth Muscle Cells ◽  
Respiratory Illnesses ◽  
The Past

Asthma is a chronic disease characterized by inflammation and hypersensitivity of airway smooth muscle cells (ASMCs) to different spasmogens. The past decade has seen increased use of herbal treatments for many chronic illnesses. Ginger ( Zingiber officinale ) is a common food plant that has been used for centuries in treating respiratory illnesses. In this study, we report the effect of its 70% aqueous methanolic crude extract (Zo·Cr) on acetylcholine (ACh)-induced airway contraction and Ca2+ signalling in ASMCs using mouse lung slices. Airway contraction and Ca2+ signalling, recorded via confocal microscopy, were induced with ACh, either alone or after pretreatment of slices with Zo·Cr and (or) verapamil, a standard Ca2+ channel blocker. ACh (10 μmol/L) stimulated airway contraction, seen as decreased airway diameter, and also stimulated Ca2+ transients (sharp rise in [Ca2+]i) and oscillations in ASMCs, seen as increased fluo-4-induced fluorescence intensity. When Zo·Cr (0.3–1.0 mg/mL) was given 30 min before ACh administration, the ACh-induced airway contraction and Ca2+ signalling were significantly reduced. Similarly, verapamil (1 μmol/L) also inhibited agonist-induced airway contraction and Ca2+ signalling, indicating a similarity in the modes of action. When Zo·Cr (0.3 mg/mL) and verapamil (1 μmol/L) were given together before ACh, the degree of inhibition was the same as that observed when each of these blockers was given alone, indicating absence of any additional inhibitory mechanism in the extract. In Ca2+-free solution, both Zo·Cr and verapamil, when given separately, inhibited Ca2+ (10 mmol/L)-induced increase in fluorescence and airway contraction. This shows that ginger inhibits airway contraction and associated Ca2+ signalling, possibly via blockade of plasma membrane Ca2+ channels, thus reiterating the effectiveness of this age-old herb in treating respiratory illnesses.

scholarly journals Single-Cell RNA Sequencing Reveals Heterogeneity of Vascular Cells in Early Stage Murine Abdominal Aortic Aneurysm—Brief Report

2021 ◽  
Vol 41 (3) ◽  
pp. 1158-1166
Author(s):  
Huan Yang ◽  
Ting Zhou ◽  
Amelia Stranz ◽  
Elise DeRoo ◽  
Bo Liu
Keyword(s):  
Smooth Muscle ◽  
Abdominal Aortic Aneurysm ◽  
Aortic Aneurysm ◽  
Smooth Muscle Cells ◽  
Single Cell ◽  
Rna Sequencing ◽  
Muscle Cells ◽  
Cell Types ◽  
Abdominal Aortic ◽  
Single Cell Rna Sequencing

Objective: Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease characterized by smooth muscle cell depletion, ECM (extracellular matrix) degradation, and infiltration of immune cells. The cellular and molecular profiles that govern the heterogeneity of the AAA aorta are yet to be elucidated. Approach and Results: We performed single-cell RNA sequencing on mouse AAA tissues. AAA was induced in C57BL/6J mice by perivascular application of CaCl 2 . Unbiased clustering identified 12 distinct populations of 8 cell types. Percentages of each population and gene expression were compared between sham and AAA tissue. Furthermore, we characterized the transcriptional profiles and potential functional features of populations in smooth muscle cells, fibroblasts, and macrophages and revealed the unique regulons in each cell type. Conclusions: Together, these data provide high-resolution insight into the complexity and heterogeneity of mouse AAA and indicate that populations within major cell types such as smooth muscle cells, fibroblasts, and macrophages may contribute differently to AAA pathogenesis. Graphic Abstract: A graphic abstract is available for this article.

Contractile agonists activate voltage-dependent calcium channels in airway smooth muscle cells

1992 ◽  
Vol 263 (1) ◽  
pp. C106-C113 ◽  
Author(s):  
M. Tomasic ◽  
J. P. Boyle ◽  
J. F. Worley ◽  
M. I. Kotlikoff
Keyword(s):  
Smooth Muscle ◽  
Calcium Channels ◽  
Smooth Muscle Cells ◽  
Calcium Channel ◽  
Airway Smooth Muscle ◽  
Muscle Cells ◽  
Cell Types ◽  
State Probability ◽  
Airway Smooth Muscle Cells ◽  
Channel Currents

To determine whether agents that cause contraction of airway smooth muscle affect sarcolemmal calcium channel activity, unitary calcium channel currents (using Ba2+ as the charge carrier) were recorded in on-cell configuration from acutely dissociated (dog, pig, and ferret) and cultured (human) airway smooth muscle cells. Addition of the contractile agonists methacholine or bradykinin increased the open-state probability of the large-conductance calcium channel 37.2- and 45-fold, respectively. The increase in open-state probability was not due to cellular depolarization because increases occurred in the absence of depolarization. Channel activation by the agonist was determined to result in the favoring of a long (16.5 +/- 5.0 ms) open lifetime for the channel, which was not observed under control conditions, in the absence of BAY K 8644. We also report the unitary calcium channel currents from a second, smaller conductance calcium channel. This channel was present in all cell types and had a mean conductance of 9.5 +/- 0.8 pS (80 mM Ba2+). Exposure of cells to agonist also resulted in an increase in the open-channel probability of the small-conductance calcium channel (10.4-fold), which did not result from cellular depolarization. These experiments demonstrate that the molecular pathways exist between contractile agonist receptors and sarcolemmal calcium channels in airway smooth muscle cells. Because membrane patches were not directly exposed to agonist, receptor-channel linkage probably occurs via a second messenger-coupling pathway.

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