scholarly journals Dual function of perivascular fibroblasts in vascular stabilization in zebrafish

PLoS Genetics ◽  
2020 ◽  
Vol 16 (10) ◽  
pp. e1008800
Author(s):  
Arsheen M. Rajan ◽  
Roger C. Ma ◽  
Katrinka M. Kocha ◽  
Dan J. Zhang ◽  
Peng Huang
Keyword(s):  
Blood Vessels ◽  
Time Lapse ◽  
Cell Types ◽  
Dual Function ◽  
Dual Roles ◽  
Vascular Integrity ◽  
Genetic Ablation ◽  
Mural Cells ◽  
And Function ◽  
Time Lapse Imaging

Blood vessels are vital to sustain life in all vertebrates. While it is known that mural cells (pericytes and smooth muscle cells) regulate vascular integrity, the contribution of other cell types to vascular stabilization has been largely unexplored. Using zebrafish, we identified sclerotome-derived perivascular fibroblasts as a novel population of blood vessel associated cells. In contrast to pericytes, perivascular fibroblasts emerge early during development, express the extracellular matrix (ECM) genes col1a2 and col5a1, and display distinct morphology and distribution. Time-lapse imaging reveals that perivascular fibroblasts serve as pericyte precursors. Genetic ablation of perivascular fibroblasts markedly reduces collagen deposition around endothelial cells, resulting in dysmorphic blood vessels with variable diameters. Strikingly, col5a1 mutants show spontaneous hemorrhage, and the penetrance of the phenotype is strongly enhanced by the additional loss of col1a2. Together, our work reveals dual roles of perivascular fibroblasts in vascular stabilization where they establish the ECM around nascent vessels and function as pericyte progenitors.

scholarly journals Dual function of perivascular fibroblasts in vascular stabilization in zebrafish

2020 ◽  
Author(s):  
Arsheen M. Rajan ◽  
Roger C. Ma ◽  
Katrinka M. Kocha ◽  
Dan J. Zhang ◽  
Peng Huang
Keyword(s):  
Extracellular Matrix ◽  
Blood Vessel ◽  
Blood Vessels ◽  
Time Lapse ◽  
Cell Types ◽  
Dual Function ◽  
Zebrafish Model ◽  
Tissue Ischemia ◽  
Genetic Ablation ◽  
Collagen Genes

ABSTRACTBlood vessels are vital to sustain life in all vertebrates. While it is known that mural cells (pericytes and smooth muscle cells) regulate vascular integrity, the contribution of other cell types to vascular stabilization has been largely unexplored. Using zebrafish, we identified sclerotome-derived perivascular fibroblasts as a novel population of blood vessel associated cells. In contrast to pericytes, perivascular fibroblasts emerge early during development, express the extracellular matrix (ECM) genes col1a2 and col5a1, and display distinct morphology and distribution. Time-lapse imaging reveals that perivascular fibroblasts serve as pericyte precursors. Genetic ablation of perivascular fibroblasts results in dysmorphic blood vessels with variable diameters. Strikingly, col5a1 mutants show spontaneous hemorrhage, and the penetrance of the phenotype is strongly enhanced by the additional loss of col1a2. Together, our work reveals dual roles of perivascular fibroblasts in vascular stabilization where they establish the ECM around nascent vessels and function as pericyte progenitors.AUTHOR SUMMARYBlood vessels are essential to sustain life in humans. Defects in blood vessels can lead to serious diseases, such as hemorrhage, tissue ischemia, and stroke. However, how blood vessel stability is maintained by surrounding support cells is still poorly understood. Using the zebrafish model, we identify a new population of blood vessel associated cells termed perivascular fibroblasts, which originate from the sclerotome, an embryonic structure that is previously known to generate the skeleton of the animal. Perivascular fibroblasts are distinct from pericytes, a known population of blood vessel support cells. They become associated with blood vessels much earlier than pericytes and express several collagen genes, encoding main components of the extracellular matrix. Loss of perivascular fibroblasts or mutations in collagen genes result in fragile blood vessels prone to damage. Using cell tracing in live animals, we find that a subset of perivascular fibroblasts can differentiate into pericytes. Together, our work shows that perivascular fibroblasts play two important roles in maintaining blood vessel integrity. Perivascular fibroblasts secrete collagens to stabilize newly formed blood vessels and a sub-population of these cells also functions as precursors to generate pericytes to provide additional vascular support.

scholarly journals A Toolbox to Characterize Human Induced Pluripotent Stem Cell–Derived Kidney Cell Types and Organoids

2019 ◽  
Vol 30 (10) ◽  
pp. 1811-1823 ◽  
Author(s):  
Jessica M. Vanslambrouck ◽  
Sean B. Wilson ◽  
Ker Sin Tan ◽  
Joanne Y.-C. Soo ◽  
Michelle Scurr ◽  
...  
Keyword(s):  
Kidney Cell ◽  
Human Kidney ◽  
Time Lapse ◽  
Cell Types ◽  
Isolation And Characterization ◽  
Gene And Protein Expression ◽  
Induced Pluripotent ◽  
Time Lapse Imaging ◽  
Kidney Morphogenesis ◽  
Kidney Organoids

BackgroundThe generation of reporter lines for cell identity, lineage, and physiologic state has provided a powerful tool in advancing the dissection of mouse kidney morphogenesis at a molecular level. Although use of this approach is not an option for studying human development in vivo, its application in human induced pluripotent stem cells (iPSCs) is now feasible.MethodsWe used CRISPR/Cas9 gene editing to generate ten fluorescence reporter iPSC lines designed to identify nephron progenitors, podocytes, proximal and distal nephron, and ureteric epithelium. Directed differentiation to kidney organoids was performed according to published protocols. Using immunofluorescence and live confocal microscopy, flow cytometry, and cell sorting techniques, we investigated organoid patterning and reporter expression characteristics.ResultsEach iPSC reporter line formed well patterned kidney organoids. All reporter lines showed congruence of endogenous gene and protein expression, enabling isolation and characterization of kidney cell types of interest. We also demonstrated successful application of reporter lines for time-lapse imaging and mouse transplantation experiments.ConclusionsWe generated, validated, and applied a suite of fluorescence iPSC reporter lines for the study of morphogenesis within human kidney organoids. This fluorescent iPSC reporter toolbox enables the visualization and isolation of key populations in forming kidney organoids, facilitating a range of applications, including cellular isolation, time-lapse imaging, protocol optimization, and lineage-tracing approaches. These tools offer promise for enhancing our understanding of this model system and its correspondence with human kidney morphogenesis.

scholarly journals The RNAseIII enzyme Drosha is critical in T cells for preventing lethal inflammatory disease

2008 ◽  
Vol 205 (9) ◽  
pp. 2005-2017 ◽  
Author(s):  
Mark M.W. Chong ◽  
Jeffrey P. Rasmussen ◽  
Alexander Y. Rudensky ◽  
Dan R. Littman
Keyword(s):  
T Cell ◽  
Inflammatory Disease ◽  
Cell Types ◽  
Mirna Biogenesis ◽  
Cell Compartment ◽  
Foxp3 Gene ◽  
Genetic Ablation ◽  
And Function ◽  
Specific Deletion

MicroRNAs (miRNAs) are implicated in the differentiation and function of many cell types. We provide genetic and in vivo evidence that the two RNaseIII enzymes, Drosha and Dicer, do indeed function in the same pathway. These have previously been shown to mediate the stepwise maturation of miRNAs (Lee, Y., C. Ahn, J. Han, H. Choi, J. Kim, J. Yim, J. Lee, P. Provost, O. Radmark, S. Kim, and V.N. Kim. 2003. Nature. 425:415–419), and genetic ablation of either within the T cell compartment, or specifically within Foxp3+ regulatory T (T reg) cells, results in identical phenotypes. We found that miRNA biogenesis is indispensable for the function of T reg cells. Specific deletion of either Drosha or Dicer phenocopies mice lacking a functional Foxp3 gene or Foxp3+ cells, whereas deletion throughout the T cell compartment also results in spontaneous inflammatory disease, but later in life. Thus, miRNA-dependent regulation is critical for preventing spontaneous inflammation and autoimmunity.

Fijiyama: a registration tool for 3D multimodal time-lapse imaging

2020 ◽  
Author(s):  
Romain Fernandez ◽  
Cédric Moisy
Keyword(s):  
Time Series Data ◽  
Time Lapse ◽  
Supplementary Information ◽  
Series Data ◽  
3D Registration ◽  
Extensive Documentation ◽  
Intractable Problems ◽  
Mri Scans ◽  
And Function ◽  
Time Lapse Imaging

Abstract Summary The increasing interest of animal and plant research communities for biomedical 3D imaging devices results in the emergence of new topics. The anatomy, structure and function of tissues can be observed non-destructively in time-lapse multimodal imaging experiments by combining the outputs of imaging devices such as X-ray CT and MRI scans. However, living samples cannot remain in these devices for a long period. Manual positioning and natural growth of the living samples induce variations in the shape, position and orientation in the acquired images that require a preprocessing step of 3D registration prior to analyses. This registration step becomes more complex when combining observations from devices that highlight various tissue structures. Identifying image invariants over modalities is challenging and can result in intractable problems. Fijiyama, a Fiji plugin built upon biomedical registration algorithms, is aimed at non-specialists to facilitate automatic alignment of 3D images acquired either at successive times and/or with different imaging systems. Its versatility was assessed on four case studies combining multimodal and time series data, spanning from micro to macro scales. Availability and implementation Fijiyama is an open source software (GPL license) implemented in Java. The plugin is available through the official Fiji release. An extensive documentation is available at the official page: https://imagej.github.io/Fijiyama Supplementary information Supplementary data are available at Bioinformatics online.

Dendritic Spines: The Locus of Structural and Functional Plasticity

2014 ◽  
Vol 94 (1) ◽  
pp. 141-188 ◽  
Author(s):  
Carlo Sala ◽  
Menahem Segal
Keyword(s):  
Dendritic Spines ◽  
Synapse Formation ◽  
Complex Structure ◽  
Time Lapse ◽  
Spiny Neurons ◽  
Synaptic Interactions ◽  
Health And Disease ◽  
Spine Structure ◽  
And Function ◽  
Time Lapse Imaging

The introduction of high-resolution time lapse imaging and molecular biological tools has changed dramatically the rate of progress towards the understanding of the complex structure-function relations in synapses of central spiny neurons. Standing issues, including the sequence of molecular and structural processes leading to formation, morphological change, and longevity of dendritic spines, as well as the functions of dendritic spines in neurological/psychiatric diseases are being addressed in a growing number of recent studies. There are still unsettled issues with respect to spine formation and plasticity: Are spines formed first, followed by synapse formation, or are synapses formed first, followed by emergence of a spine? What are the immediate and long-lasting changes in spine properties following exposure to plasticity-producing stimulation? Is spine volume/shape indicative of its function? These and other issues are addressed in this review, which highlights the complexity of molecular pathways involved in regulation of spine structure and function, and which contributes to the understanding of central synaptic interactions in health and disease.

scholarly journals Identification of astroglia-like cardiac nexus glia that are critical regulators of cardiac development and function

PLoS Biology ◽  
2021 ◽  
Vol 19 (11) ◽  
pp. e3001444
Author(s):  
Nina L. Kikel-Coury ◽  
Jacob P. Brandt ◽  
Isabel A. Correia ◽  
Michael R. O’Dea ◽  
Dana F. DeSantis ◽  
...  
Keyword(s):  
Heart Rate ◽  
Nervous System ◽  
Cardiac Development ◽  
Time Lapse ◽  
Zebrafish Model ◽  
Single Cell Sequencing ◽  
Parasympathetic System ◽  
And Function ◽  
Time Lapse Imaging

Glial cells are essential for functionality of the nervous system. Growing evidence underscores the importance of astrocytes; however, analogous astroglia in peripheral organs are poorly understood. Using confocal time-lapse imaging, fate mapping, and mutant genesis in a zebrafish model, we identify a neural crest–derived glial cell, termed nexus glia, which utilizes Meteorin signaling via Jak/Stat3 to drive differentiation and regulate heart rate and rhythm. Nexus glia are labeled with gfap, glast, and glutamine synthetase, markers that typically denote astroglia cells. Further, analysis of single-cell sequencing datasets of human and murine hearts across ages reveals astrocyte-like cells, which we confirm through a multispecies approach. We show that cardiac nexus glia at the outflow tract are critical regulators of both the sympathetic and parasympathetic system. These data establish the crucial role of glia on cardiac homeostasis and provide a description of nexus glia in the PNS.

scholarly journals Achieving functional neuronal dendrite structure through sequential stochastic growth and retraction

2020 ◽  
Author(s):  
André Ferreira Castro ◽  
Lothar Baltruschat ◽  
Tomke Stürner ◽  
Amirhoushang Bahrami ◽  
Peter Jedlicka ◽  
...  
Keyword(s):  
Body Wall ◽  
Time Lapse ◽  
Membrane Curvature ◽  
Class I ◽  
List Type ◽  
Larval Body ◽  
And Function ◽  
Dendritic Branches ◽  
Time Lapse Imaging

AbstractClass I ventral posterior dendritic arborisation (c1vpda) proprioceptive sensory neurons respond to contractions in the Drosophila larval body wall during crawling. Their dendritic branches run along the direction of contraction, possibly a functional requirement to maximise membrane curvature during crawling contractions. Although the molecular machinery of dendritic patterning in c1vpda has been extensively studied, the process leading to the precise elaboration of their comb-like shapes remains elusive. Here, to link dendrite shape with its proprioceptive role, we performed long-term, non-invasive, in vivo time-lapse imaging of c1vpda embryonic and larval morphogenesis to reveal a sequence of differentiation stages. We combined computer models and dendritic branch dynamics tracking to propose that distinct sequential phases of targeted growth and stochastic retraction achieve efficient dendritic trees both in terms of wire and function. Our study shows how dendrite growth balances structure–function requirements, shedding new light on general principles of self-organisation in functionally specialised dendrites.In briefAn optimal wire and function trade-off emerges from noisy growth and stochastic retraction during Drosophila class I ventral posterior dendritic arborisation (c1vpda) dendrite development.HighlightsC1vpda dendrite outgrowth follows wire constraints.Stochastic retraction of functionally suboptimal branches in a subsequent growth phase.C1vpda growth rules favour branches running parallel to larval body wall contraction.Comprehensive growth model reproduces c1vpda development in silico.

scholarly journals JMJD6 Dysfunction Due to Iron Deficiency in Preeclampsia Disrupts Fibronectin Homeostasis Resulting in Diminished Trophoblast Migration

2021 ◽  
Vol 9 ◽  
Author(s):  
Sruthi Alahari ◽  
Abby Farrell ◽  
Leonardo Ermini ◽  
Chanho Park ◽  
Julien Sallais ◽  
...  
Keyword(s):  
Time Lapse ◽  
Lysosomal Degradation ◽  
Related Disorder ◽  
Novel Target ◽  
And Function ◽  
Time Lapse Imaging ◽  
Fibronectin Assembly ◽  
Degradation Time ◽  
Trophoblast Migration

The mechanisms contributing to excessive fibronectin in preeclampsia, a pregnancy-related disorder, remain unknown. Herein, we investigated the role of JMJD6, an O2- and Fe2+-dependent enzyme, in mediating placental fibronectin processing and function. MALDI-TOF identified fibronectin as a novel target of JMJD6-mediated lysyl hydroxylation, preceding fibronectin glycosylation, deposition, and degradation. In preeclamptic placentae, fibronectin accumulated primarily in lysosomes of the mesenchyme. Using primary placental mesenchymal cells (pMSCs), we found that fibronectin fibril formation and turnover were markedly impeded in preeclamptic pMSCs, partly due to impaired lysosomal degradation. JMJD6 knockdown in control pMSCs recapitulated the preeclamptic FN phenotype. Importantly, preeclamptic pMSCs had less total and labile Fe2+ and Hinokitiol treatment rescued fibronectin assembly and promoted lysosomal degradation. Time-lapse imaging demonstrated that defective ECM deposition by preeclamptic pMSCs impeded HTR-8/SVneo cell migration, which was rescued upon Hinokitiol exposure. Our findings reveal new Fe2+-dependent mechanisms controlling fibronectin homeostasis/function in the placenta that go awry in preeclampsia.

scholarly journals CaM kinase Iα–induced phosphorylation of Drp1 regulates mitochondrial morphology

2008 ◽  
Vol 182 (3) ◽  
pp. 573-585 ◽  
Author(s):  
Xiao-Jian Han ◽  
Yun-Fei Lu ◽  
Shun-Ai Li ◽  
Taku Kaitsuka ◽  
Yasufumi Sato ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Mitochondrial Dynamics ◽  
Mitochondrial Fission ◽  
Time Lapse ◽  
Cell Types ◽  
Mitochondrial Morphology ◽  
Voltage Dependent ◽  
Dependent Protein ◽  
Time Lapse Imaging

Mitochondria are dynamic organelles that frequently move, divide, and fuse with one another to maintain their architecture and functions. However, the signaling mechanisms involved in these processes are still not well characterized. In this study, we analyze mitochondrial dynamics and morphology in neurons. Using time-lapse imaging, we find that Ca2+ influx through voltage-dependent Ca2+ channels (VDCCs) causes a rapid halt in mitochondrial movement and induces mitochondrial fission. VDCC-associated Ca2+ signaling stimulates phosphorylation of dynamin-related protein 1 (Drp1) at serine 600 via activation of Ca2+/calmodulin-dependent protein kinase Iα (CaMKIα). In neurons and HeLa cells, phosphorylation of Drp1 at serine 600 is associated with an increase in Drp1 translocation to mitochondria, whereas in vitro, phosphorylation of Drp1 results in an increase in its affinity for Fis1. CaMKIα is a widely expressed protein kinase, suggesting that Ca2+ is likely to be functionally important in the control of mitochondrial dynamics through regulation of Drp1 phosphorylation in neurons and other cell types.

scholarly journals Achieving functional neuronal dendrite structure through sequential stochastic growth and retraction

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
André Ferreira Castro ◽  
Lothar Baltruschat ◽  
Tomke Stürner ◽  
Amirhoushang Bahrami ◽  
Peter Jedlicka ◽  
...  
Keyword(s):  
Time Lapse ◽  
Membrane Curvature ◽  
Larval Body ◽  
Stochastic Growth ◽  
Molecular Machinery ◽  
Neuronal Dendrite ◽  
And Function ◽  
Dendritic Branches ◽  
Time Lapse Imaging

Class I ventral posterior dendritic arborisation (c1vpda) proprioceptive sensory neurons respond to contractions in the Drosophila larval body wall during crawling. Their dendritic branches run along the direction of contraction, possibly a functional requirement to maximise membrane curvature during crawling contractions. Although the molecular machinery of dendritic patterning in c1vpda has been extensively studied, the process leading to the precise elaboration of their comb-like shapes remains elusive. Here, to link dendrite shape with its proprioceptive role, we performed long-term, non-invasive, in vivo time-lapse imaging of c1vpda embryonic and larval morphogenesis to reveal a sequence of differentiation stages. We combined computer models and dendritic branch dynamics tracking to propose that distinct sequential phases of stochastic growth and retraction achieve efficient dendritic trees both in terms of wire and function. Our study shows how dendrite growth balances structure–function requirements, shedding new light on general principles of self-organisation in functionally specialised dendrites.

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