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Intravital microscopy

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https://www.readbyqxmd.com/read/28428216/maturation-of-platelet-function-during-murine-fetal-development-in-vivo
#1
Andreas Margraf, Claudia Nussbaum, Ina Rohwedder, Sarah Klapproth, Angela R M Kurz, Annamaria Florian, Volker Wiebking, Joachim Pircher, Monika Pruenster, Roland Immler, Steffen Dietzel, Ludmila Kremer, Friedemann Kiefer, Markus Moser, Andreas W Flemmer, Elizabeth Quackenbush, Ulrich H von Andrian, Markus Sperandio
OBJECTIVE: Platelet function has been intensively studied in the adult organism. However, little is known about the function and hemostatic capacity of platelets in the developing fetus as suitable in vivo models are lacking. APPROACH AND RESULTS: To examine fetal platelet function in vivo, we generated a fetal thrombosis model and investigated light-/dye-induced thrombus formation by intravital microscopy throughout gestation. We observed that significantly less and unstable thrombi were formed at embryonic day (E) 13...
April 20, 2017: Arteriosclerosis, Thrombosis, and Vascular Biology
https://www.readbyqxmd.com/read/28416692/immunomodulation-accelerated-neuronal-regeneration-following-selective-rod-photoreceptor-cell-ablation-in-the-zebrafish-retina
#2
David T White, Sumitra Sengupta, Meera T Saxena, Qingguo Xu, Justin Hanes, Ding Ding, Hongkai Ji, Jeff S Mumm
Müller glia (MG) function as inducible retinal stem cells in zebrafish, completely repairing the eye after damage. The innate immune system has recently been shown to promote tissue regeneration in which classic wound-healing responses predominate. However, regulatory roles for leukocytes during cellular regeneration-i.e., selective cell-loss paradigms akin to degenerative disease-are less well defined. To investigate possible roles innate immune cells play during retinal cell regeneration, we used intravital microscopy to visualize neutrophil, macrophage, and retinal microglia responses to induced rod photoreceptor apoptosis...
April 17, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28413812/quantitative-3d-time-lapse-imaging-of-muscle-progenitors-in-skeletal-muscle-of-live-mice
#3
Micah T Webster, Tyler Harvey, Chen-Ming Fan
For non-optically clear mammalian tissues, it is now possible to use multi-photon microscopy to penetrate deep into the tissue and obtain detailed single cell images in a live animal, i.e., intravital imaging. This technique is in principle applicable to any fluorescently marked cell, and we have employed it to observe stem cells during the regenerative process. Stem cell-mediated skeletal muscle regeneration in the mouse model has been classically studied at specific time points by sacrificing the animal and harvesting the muscle tissue for downstream analyses...
December 20, 2016: Bio-protocol
https://www.readbyqxmd.com/read/28410977/intravital-imaging-of-the-kidney
#4
REVIEW
Takashi Hato, Seth Winfree, Pierre C Dagher
Two-photon intravital microscopy is a powerful tool that allows the examination of dynamic cellular processes in the live animal with unprecedented resolution. Indeed, it offers the ability to address unique biological questions that may not be solved by other means. While two-photon intravital microscopy has been successfully applied to study many organs, the kidney presents its own unique challenges that need to be overcome in order to optimize and validate imaging data. For kidney imaging, the complexity of renal architecture and salient autofluorescence merit special considerations as these elements directly impact image acquisition and data interpretation...
April 11, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28405670/a-long-acting-pai-1-inhibitor-reduce-thrombus-formation
#5
Shuangzhou Peng, Guangpu Xue, Lihu Gong, Chao Fang, Jingfei Chen, Cai Yuan, Zhuo Chen, Lishan Yao, Bruce Furie, Mingdong Huang
Plasminogen activator inhibitor 1 (PAI-1) is the main inhibitor of tissue-type and urokinase-type plasminogen activators (t/uPA) and plays an important role in the fibrinolytic system. Inhibition of PAI-1 activity prevents thrombosis and accelerates fibrinolysis, indicating that PAI-1 inhibitors may be used as effective antithrombotic agents. We previously designed a PAI-1 inhibitor (PAItrap) which is a variant of inactivated urokinase protease domain. In the present study, we tried to fuse PAItrap with human serum albumin (HSA) to develop a long-acting PAI-1 inhibitor...
April 13, 2017: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/28393199/smad4-deletion-in-blood-vessel-endothelial-cells-promotes-ovarian-cancer-metastasis
#6
Jie Yang, Ya Wang, Zhen Zeng, Long Qiao, Liang Zhuang, Qinglei Gao, Ding Ma, Xiaoyuan Huang
SMAD4 is a critical co-smad in signal transduction pathways activated in response to transforming growth factor-β (TGF-β)-related ligands, regulating cell growth and differentiation. The roles played by SMAD4 inactivation in tumors highlighted it as a tumor-suppressor gene. Herein, we report that loss of SMAD4 expression in vascular endothelial cells promotes ovarian cancer invasion. SiRNA transfer of this gene in the HUVEC reduced SMAD4 protein expression and function. Although it reduced the vessel endothelial cell tubule formation in vitro and in vivo, it did not affect the tumor growth significantly in vivo...
May 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/28391344/is-there-a-role-for-tnf%C3%AE-blockade-in-anca-associated-vasculitis-and-glomerulonephritis
#7
Stephen P McAdoo, Charles D Pusey
Tumour necrosis factor alpha (TNFα) is a cytokine that is pivotal in the inflammatory response. Blockade of TNFα has been shown to be effective in a number of human autoimmune diseases, including rheumatoid arthritis, raising the question of whether this approach may be effective in inflammatory kidney disease, such as ANCA-associated vasculitis (AAV). In AAV, there is considerable evidence for the role of TNFα in the pathophysiology of disease, including increased expression of TNFα mRNA in leucocytes and in renal tissue...
January 1, 2017: Nephrology, Dialysis, Transplantation
https://www.readbyqxmd.com/read/28385784/the-endothelin-b-receptor-plays-a-crucial-role-for-the-adhesion-of-neutrophils-to-the-endothelium-in-sickle-cell-disease
#8
Bérengère Koehl, Pierre Nivoit, Wassim El Nemer, Olivia Lenoir, Patricia Hermand, Catia Pereira, Valentine Brousse, Léa Guyonnet, Giulia Ghinatti, Malika Benkerrou, Yves Colin, Caroline Le Van Kim, Pierre-Louis Tharaux
Although the primary origin of sickle cell disease is a hemoglobin disorder, several cell types contribute considerably to the physiopathology of the disease. The adhesion of neutrophils to activated endothelium is critical in sickle cell disease pathophysiology and the targeting of neutrophils and their interactions with endothelium represent important opportunities for new therapeutics. We focused on endothelin-1, a mediator involved in neutrophil activation and recruitment in tissues, and we investigated the involvement of the endothelin receptors in interaction of neutrophils with endothelial cells...
April 6, 2017: Haematologica
https://www.readbyqxmd.com/read/28385296/17%C3%AE-estradiol-prevents-mesenteric-injury-induced-by-occlusion-of-the-proximal-descending-aorta-in-male-rats
#9
Paulo Thales Rocha de Sousa, Ana Cristina Breithaupt-Faloppa, Cristiano de Jesus Correia, Raif Restivo Simão, Sueli Gomes Ferreira, Alfredo Inácio Fiorelli, Luiz Felipe Pinho Moreira, Paulina Sannomiya
OBJECTIVE: In surgical aortic repair or cardiac surgery with aorta occlusion, the occurrence of mesenteric ischemia and bowel injury has been associated with higher short-term mortality. The vascular protection of estrogens has been investigated and is mainly mediated by increasing the availability of nitric oxide (NO). Therefore, this study investigated the role of 17β-estradiol on visceral ischemia-reperfusion (I/R) injury after descending aorta occlusion in male rats. METHODS: Mesenteric ischemia was induced in male Wistar rats by placing a 2F Fogarty arterial embolectomy catheter (Edwards Lifesciences, Irvine, Calif) in the descending aorta, which remained occluded for 15 minutes, followed by reperfusion for up to 2 hours...
April 3, 2017: Journal of Vascular Surgery
https://www.readbyqxmd.com/read/28382036/in-vivo-imaging-sheds-light-on-immune-cell-migration-and-function-in-cancer
#10
REVIEW
Tommaso Torcellan, Jessica Stolp, Tatyana Chtanova
There is ample evidence for both beneficial and harmful involvement of the immune system in tumor development and spread. Immune cell recruitment to tumors is essential not only for the success of anticancer immune therapies but also for tumor-induced immune suppression. Now that immune-based therapies are playing an increasingly important role in treatment of solid tumors such as metastatic melanomas, precise analysis of the in vivo contributions of different leukocyte subsets in tumor immunity has become an even greater priority...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28378876/potentials-and-limitations-of-integra%C3%A2-flowable-wound-matrix-seeded-with-adipose-tissue-derived-microvascular-fragments
#11
T Später, F S Frueh, M D Menger, M W Laschke
Adipose tissue-derived microvascular fragments (ad-MVF) represent promising vascularisation units for bioengineered Integra® matrix wound dressing (MWD). However, due to the sheet-like structure with small pore sizes, the seeding of this matrix with ad-MVF is mainly limited to its surface. Integra® flowable wound matrix (FWM) may be suitable to achieve a more homogeneous distribution and, thus, improved vascularisation, because this gel-like matrix allows for the direct admixture of ad-MVF during sample preparation...
April 5, 2017: European Cells & Materials
https://www.readbyqxmd.com/read/28377486/p2x7-deficiency-blocks-lesional-inflammasome-activity-and-ameliorates-atherosclerosis-in-mice
#12
Peter Stachon, Adrian Heidenreich, Julian Merz, Ingo Hilgendorf, Dennis Wolf, Florian Willecke, Sunaina von Garlen, Philipp Albrecht, Carmen Härdtner, Nicolas Ehrat, Natalie Hoppe, Jochen Reinöhl, Constantin von Zur Mühlen, Christoph Bode, Marco Idzko, Andreas Zirlik
Background -Extracellular ATP binds as a danger signal to purinergic receptor P2X7, promotes inflammasome assembly, and IL-1β expression. We hypothesized a functional role of the signal axis ATP - P2X7 in inflammasome activation and the chronic inflammation driving atherosclerosis. Methods -P2X7-competent and -deficient macrophages were isolated and stimulated with LPS, ATP, or both. To assess whether P2X7 may have a role in atherosclerosis, P2X7 expression was analyzed in aortic arches from LDLR(-/-) mice consuming high cholesterol diet (HCD) or chow diet...
April 4, 2017: Circulation
https://www.readbyqxmd.com/read/28369007/drug-induced-defibrinogenation-as-new-treatment-approach-of-acute-hearing-loss-in-an-animal-model-for-inner-ear-vascular-impairment
#13
Bernhard G Weiss, Mattis Bertlich, Stephan A Bettag, Hendrik Desinger, Friedrich Ihler, Martin Canis
OBJECTIVE: Disturbance of cochlear microcirculation is considered to be the final common pathway of various inner ear diseases. Hyperfibrinogenemia causing increased plasma viscosity is a known risk factor for sudden sensorineural hearing loss and may lead to a critical reduction of cochlear blood flow. The aim of this study was to evaluate the effect of a substantial reduction of plasma fibrinogen levels by drug-induced defibrinogenation for the treatment of acute hearing loss in vivo...
March 31, 2017: Otology & Neurotology
https://www.readbyqxmd.com/read/28366932/intravital-microscopy-in-historic-and-contemporary-immunology
#14
REVIEW
Judith Secklehner, Cristina Lo Celso, Leo M Carlin
In this review, we discuss intravital microscopy of immune cells, starting from its historic origins to current applications in diverse organs. It is clear from a quantitative review of the literature that intravital microscopy is a key tool in both historic and contemporary immunological research, providing unique advances in our understanding of immune responses. We have chosen to focus this review on how intravital microscopy methodologies are used to image specific organs or systems and we present recent descriptions of fundamental immunological processes that could not have been achieved by other methods...
April 3, 2017: Immunology and Cell Biology
https://www.readbyqxmd.com/read/28363496/therapeutic-progenitor-cell-application-for-tissue-regeneration-analyzing-the-impact-of-toll-like-receptor-signaling-on-c-kit-cell-migration-following-ischemia-reperfusion-injury-in-vivo
#15
Peter Donndorf, Saifullah Abubaker, Brigitte Vollmar, Christian Rimmbach, Gustav Steinhoff, Alexander Kaminski
OBJECTIVES: Toll-like-receptor (TLR) mediated immune response has been shown to regulate myocardial damage following cardiac ischemia-reperfusion (IR). It has not been described conclusively so far whether migration of therapeutically applied progenitor cells following an IR event depends on TLR-signaling. METHODS: In vivo migratory capacity murine c-kit(+) cells following IR injury was quantified by intravital fluorescence microscopy, utilizing the mouse cremaster muscle model and analyzing early (rolling) and late (adhesion) c-kit(+) cell interaction with the local endothelium...
March 29, 2017: Microvascular Research
https://www.readbyqxmd.com/read/28362378/combining-intravital-fluorescent-microscopy-ivfm-with-genetic-models-to-study-engraftment-dynamics-of-hematopoietic-cells-to-bone-marrow-niches
#16
Lin Wang, Malgorzata M Kamocka, Amy Zollman, Nadia Carlesso
Increasing evidence indicates that normal hematopoiesis is regulated by distinct microenvironmental cues in the BM, which include specialized cellular niches modulating critical hematopoietic stem cell (HSC) functions(1)(,)(2). Indeed, a more detailed picture of the hematopoietic microenvironment is now emerging, in which the endosteal and the endothelial niches form functional units for the regulation of normal HSC and their progeny(3)(,)(4)(,)(5). New studies have revealed the importance of perivascular cells, adipocytes and neuronal cells in maintaining and regulating HSC function(6)(,)(7)(,)(8)...
March 21, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28362362/combined-intravital-microscopy-and-contrast-enhanced-ultrasonography-of-the-mouse-hindlimb-to-study-insulin-induced-vasodilation-and-muscle-perfusion
#17
Alexander H Turaihi, Erik M van Poelgeest, Victor W M van Hinsbergh, Erik H Serné, Yvo M Smulders, Etto C Eringa
It has been demonstrated that insulin's vascular actions contribute to regulation of insulin sensitivity. Insulin's effects on muscle perfusion regulate postprandial delivery of nutrients and hormones to insulin-sensitive tissues. We here describe a technique for combining intravital microscopy (IVM) and contrast-enhanced ultrasonography (CEUS) of the adductor compartment of the mouse hindlimb to simultaneously visualize muscle resistance arteries and perfusion of the microcirculation in vivo. Simultaneously assessing insulin's effect at multiple levels of the vascular tree is important to study relationships between insulin's multiple vasoactive effects and muscle perfusion...
March 20, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28356517/protectin-d1n-3-dpa-and-resolvin-d5n-3-dpa-are-effectors-of-intestinal-protection
#18
Thomas Gobbetti, Jesmond Dalli, Romain A Colas, Donata Federici Canova, Marius Aursnes, Delphine Bonnet, Laurent Alric, Nathalie Vergnolle, Celine Deraison, Trond V Hansen, Charles N Serhan, Mauro Perretti
The resolution of inflammation is an active process orchestrated by specialized proresolving lipid mediators (SPM) that limit the host response within the affected tissue; failure of effective resolution may lead to tissue injury. Because persistence of inflammatory signals is a main feature of chronic inflammatory conditions, including inflammatory bowel diseases (IBDs), herein we investigate expression and functions of SPM in intestinal inflammation. Targeted liquid chromatography-tandem mass spectrometry-based metabololipidomics was used to identify SPMs from n-3 polyunsaturated fatty acids in human IBD colon biopsies, quantifying a significant up-regulation of the resolvin and protectin pathway compared with normal gut tissue...
March 29, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28351758/the-mouse-cortical-meninges-are-the-site-of-immune-responses-to-many-different-pathogens-and-are-accessible-to-in-vivo-imaging
#19
REVIEW
Jonathan A Coles, Phillip J Stewart-Hutchinson, Elmarie Myburgh, James M Brewer
A wide range of viral and microbial infections are known to cause meningitis, and there is evidence that the meninges are the gateway to pathogenic invasion of the brain parenchyma. Hence observation of these regions has wide application to understanding host-pathogen interactions. Interactions between pathogens and cells of the immune response can be modified by changes in their environment, such as suppression of the flow of blood and lymph, and, particularly in the case of the meninges with their unsupported membranes, invasive dissection can alter the tissue architecture...
March 25, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28349487/t-cell-response-in-the-lung-following-influenza-virus-infection
#20
Robert A Benson, Jennifer C Lawton, Megan K L MacLeod
Methods that enable the identification of virus-specific CD4 and CD8 T cells are key to our understanding of how the adaptive immune response controls viral infection. Here we describe two distinct methods to evaluate the T cell response to influenza A virus (IAV). The number and phenotype of T cells that respond to natural IAV epitopes can be assessed by flow cytometry using MHC class I and class II tetramers. Using this system, IAV-specific T cells can be tracked in various organs within the same animal, or, in different cohorts, the response can be evaluated at several time points following infection...
2017: Methods in Molecular Biology
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