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Intravital microscopy

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https://www.readbyqxmd.com/read/28811547/up-regulation-of-nadph-oxidase-mediated-redox-signaling-contributes-to-the-loss-of-barrier-function-in-krit1-deficient-endothelium
#1
Luca Goitre, Peter V DiStefano, Andrea Moglia, Nicholas Nobiletti, Eva Baldini, Lorenza Trabalzini, Julie Keubel, Eliana Trapani, Vladimir V Shuvaev, Vladimir R Muzykantov, Ingrid H Sarelius, Saverio Francesco Retta, Angela J Glading
The intracellular scaffold KRIT1/CCM1 is an established regulator of vascular barrier function. Loss of KRIT1 leads to decreased microvessel barrier function and to the development of the vascular disorder Cerebral Cavernous Malformation (CCM). However, how loss of KRIT1 causes the subsequent deficit in barrier function remains undefined. Previous studies have shown that loss of KRIT1 increases the production of reactive oxygen species (ROS) and exacerbates vascular permeability triggered by several inflammatory stimuli, but not TNF-α...
August 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28810287/seeding-density-is-a-crucial-determinant-for-the-in-vivo-vascularisation-capacity-of-adipose-tissue-derived-microvascular-fragments
#2
T Später, C Körbel, F S Frueh, R M Nickels, M D Menger, M W Laschke
Adipose tissue-derived microvascular fragments (ad-MVF) represent effective vascularisation units for the seeding of dermal substitutes. However, particularly in case of extensive skin defects, the required amounts of donor fat tissue for the harvesting of ad-MVF may not always be available. Therefore, we herein determined the lowest ad-MVF density needed to induce a sufficient vascularisation and incorporation of seeded implants. Collagen-glycosaminoglycan matrices (Integra®; diameter: 4 mm) were seeded with 15,000 (HD), 10,000 (MD) and 5,000 (LD) ad-MVF and implanted into full-thickness skin defects within mouse dorsal skinfold chambers, to analyse their in vivo vascularisation and incorporation...
August 15, 2017: European Cells & Materials
https://www.readbyqxmd.com/read/28806702/hmgb1-redox-during-sepsis
#3
Wasan Abdulmahdi, Devika Patel, May M Rabadi, Tala Azar, Edson Jules, Mark Lipphardt, Rameen Hashemiyoon, Brian B Ratliff
During sepsis, the alarmin HMGB1 is released from tissues and promotes systemic inflammation that results in multi-organ damage, with the kidney particularly susceptible to injury. The severity of inflammation and pro-damage signaling mediated by HMGB1 appears to be dependent on the alarmin's redox state. Therefore, we examined HMGB1 redox in kidney cells during sepsis. Using intravital microscopy, CellROX labeling of kidneys in live mice indicated increased ROS generation in the kidney perivascular endothelium and tubules during lipopolysaccharide (LPS)-induced sepsis...
August 4, 2017: Redox Biology
https://www.readbyqxmd.com/read/28802564/interactions-between-platelets-and-inflammatory-monocytes-affect-sickness-behavior-in-mice-with-liver-inflammation
#4
Charlotte D'Mello, Wagdi Almishri, Hongqun Liu, Mark Gordon Swain
BACKGROUND & AIMS: Patients with inflammatory liver disease commonly develop debilitating symptoms, called sickness behaviors, which arise via changes in brain function. Monocytes that produce tumor necrosis factor (TNF) interact with cerebral endothelial cells to activate microglial cells and promote sickness behavior. Platelets regulate inflammation, and aggregates of monocytes and platelets are increased in the circulation of patients with liver disease. We investigated the role of platelets in inducing inflammatory features of circulating monocytes and promoting sickness behaviors in mice with cholestatic liver injury...
August 9, 2017: Gastroenterology
https://www.readbyqxmd.com/read/28800593/in-vivo-functional-and-morphological-characterization-of-bone-and-striated-muscle-microcirculation-in-nsg-mice
#5
Haider Mussawy, Lennart Viezens, Gerrit Hauenherm, Malte Schroeder, Christian Schaefer
Organ-specific microcirculation plays a central role in tumor growth, tumor cell homing, tissue engineering, and wound healing. Mouse models are widely used to study these processes; however, these mouse strains often possess unique microhemodynamic parameters, making it difficult to directly compare experiments. The full functional characterization of bone and striated muscle microcirculatory parameters in non-obese diabetic-severe combined immunodeficiency/y-chain; NOD-Prkds IL2rg (NSG) mice has not yet been reported...
2017: PloS One
https://www.readbyqxmd.com/read/28798204/imaging-igf-i-uptake-in-growth-plate-cartilage-using-in-vivo-multiphoton-microscopy
#6
Maria A Serrat, Gabriela Ion
Bones elongate through endochondral ossification in cartilaginous growth plates located at ends of primary long bones. Linear growth ensues from a cascade of biochemical signals initiated by actions of systemic and local regulators on growth plate chondrocytes. Although cellular processes are well defined, there is a fundamental gap in understanding how growth regulators are physically transported from surrounding blood vessels into and through dense, avascular cartilage matrix. Intravital imaging using in vivo multiphoton microscopy is one promising strategy to overcome this barrier by quantitatively tracking molecular delivery to cartilage from the vasculature in real time...
August 10, 2017: Journal of Applied Physiology
https://www.readbyqxmd.com/read/28798084/a-small-population-of-liver-endothelial-cells-undergoes-endothelial-to-mesenchymal-transition-in-response-to-chronic-liver-injury
#7
Jordi Ribera, Montse Pauta, Pedro Melgar-Lesmes, Bernat Cordoba, Anna Bosch, Maria Calvo, Daniel Rodrigo-Torres, Pau Sancho-Bru, Aurea Mira, Wladimiro Jimenez, Manuel Morales-Ruiz
BACKGROUND AND AIMS: Rising evidence points to endothelial-to-mesenchymal transition (EndMT) as a significant source of the mesenchymal cell population in fibrotic diseases. In this context, we hypothesized that liver endothelial cells undergo EndMT during fibrosis progression. METHODS: Cirrhosis in mice was induced by CCl4 A transgenic mouse expressing a red fluorescent protein reporter under the control of Tie2 promoter (Tie2-tdTomato) was used to trace the acquisition of EndMT...
August 10, 2017: American Journal of Physiology. Gastrointestinal and Liver Physiology
https://www.readbyqxmd.com/read/28793186/an-engineered-optogenetic-switch-for-spatiotemporal-control-of-gene-expression-cell-differentiation-and-tissue-morphogenesis
#8
Lauren Polstein, Mark Juhas, Gabi Hanna, Nenad Bursac, Charles A Gersbach
The precise spatial and temporal control of gene expression, cell differentiation, and tissue morphogenesis has widespread application in regenerative medicine and the study of tissue development. In this work, we applied optogenetics to control cell differentiation and new tissue formation. Specifically, we engineered an optogenetic "on" switch that provides permanent transgene expression following a transient dose of blue light illumination. To demonstrate its utility in controlling cell differentiation and reprogramming, we incorporated an engineered form of the master myogenic factor MyoD into this system in multipotent cells...
August 9, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28792428/influences-of-macrohemodynamic-conditions-on-systemic-microhemodynamic-changes-in-burns
#9
Jochen-Frederick Hernekamp, Florian Neubrech, Tomke Cordts, Volker J Schmidt, Ulrich Kneser, Thomas Kremer
PURPOSE: The clinical course after major burns is characterized by microcirculatory changes and consecutive capillary leakage. However, current clinical monitoring does not properly assess microcirculation, whereas macrohemodynamic changes are continuously evaluated. Here, we assess if macrohemodynamic and microhemodynamic parameters after burn trauma are correlated in a rat model. METHODS: Burn plasma harvested from donor rats 4 hours after thermal injury (30% total body surface area, 100 °C water, 12 seconds) was administered intravenously to healthy animals during 2 hours of intravital microscopy (burn group [BG])...
November 2016: Annals of Plastic Surgery
https://www.readbyqxmd.com/read/28790339/preventing-inflammation-inhibits-biopsy-mediated-changes-in-tumor-cell-behavior
#10
Maria Alieva, Andreia S Margarido, Tamara Wieles, Erik R Abels, Burcin Colak, Carla Boquetale, Herke Jan Noordmans, Tom J Snijders, Marike L Broekman, Jacco van Rheenen
Although biopsies and tumor resection are prognostically beneficial for glioblastomas (GBM), potential negative effects have also been suggested. Here, using retrospective study of patients and intravital imaging of mice, we identify some of these negative aspects, including stimulation of proliferation and migration of non-resected tumor cells, and provide a strategy to prevent these adverse effects. By repeated high-resolution intravital microscopy, we show that biopsy-like injury in GBM induces migration and proliferation of tumor cells through chemokine (C-C motif) ligand 2 (CCL-2)-dependent recruitment of macrophages...
August 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28789683/voluntary-running-exercise-protects-against-sepsis-induced-early-inflammatory-and-pro-coagulant-responses-in-aged-mice
#11
Karel Tyml, Scott Swarbreck, Cynthia Pape, Dan Secor, James Koropatnick, Qingping Feng, Ruud A W Veldhuizen, Sean E Gill
BACKGROUND: Despite many animal studies and clinical trials, mortality in sepsis remains high. This may be due to the fact that most experimental studies of sepsis employ young animals, whereas the majority of septic patients are elderly (60 - 70 years). The objective of the present study was to examine the sepsis-induced inflammatory and pro-coagulant responses in aged mice. Since running exercise protects against a variety of diseases, we also examined the effect of voluntary running on septic responses in aged mice...
August 8, 2017: Critical Care: the Official Journal of the Critical Care Forum
https://www.readbyqxmd.com/read/28780320/non-invasive-intravital-imaging-of-head-and-neck-squamous-cell-carcinomas-in-live-mice
#12
Panomwat Amornphimoltham, Jamie Thompson, Nicolas Melis, Roberto Weigert
Head and neck squamous cell carcinoma is one of the most common cancers with a 50% 5-year survival rate. Understanding the mechanisms that control development, progression, and spreading of the tumor to distal sites is of paramount importance to develop effective therapies. Here, we describe a minimally invasive procedure, which enables performing intravital microscopy of the mouse tongue in models for oral cancer and provides structural and dynamic information of the tumors at cellular and subcellular level...
August 2, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28765364/filoquant-reveals-increased-filopodia-density-during-breast-cancer-progression
#13
Guillaume Jacquemet, Ilkka Paatero, Alexandre F Carisey, Artur Padzik, Jordan S Orange, Hellyeh Hamidi, Johanna Ivaska
Defective filopodia formation is linked to pathologies such as cancer, wherein actively protruding filopodia, at the invasive front, accompany cancer cell dissemination. Despite wide biological significance, delineating filopodia function in complex systems remains challenging and is particularly hindered by lack of compatible methods to quantify filopodia properties. Here, we present FiloQuant, a freely available ImageJ plugin, to detect filopodia-like protrusions in both fixed- and live-cell microscopy data...
August 1, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28762564/capsaicin-induced-rapid-neutrophil-leukocyte-activation-in-the-rat-urinary-bladder-microcirculatory-bed
#14
Péter Járomi, Dénes Garab, Petra Hartmann, Dóra Bodnár, Sándor Nyíri, Péter Sántha, Mihály Boros, Gábor Jancsó, Andrea Szabó
AIMS: This study was initiated to investigate the involvement of neutrophil leukocyte activation in neurogenic inflammation, a process also involved in human urinary pathologies, elicited in the rat urinary bladder by the local administration of capsaicin, the archetypal TRPV1 agonist. The contribution of afferent nerves and sensory neuropeptides to leukocyte activation in the urinary bladder microcirculatory bed was examined. METHODS: Following a 15-min topical application of capsaicin (50 μM), leukocyte-endothelial interactions were examined for an observation period of 45 min with intravital microscopy...
August 1, 2017: Neurourology and Urodynamics
https://www.readbyqxmd.com/read/28745543/enhanced-adhesion-of-blood-platelets-to-intact-endothelium-of-mesenteric-vascular-bed-in-mice-with-streptozotocin-induced-diabetes-is-mediated-by-an-up-regulated-endothelial-surface-deposition-of-vwf-in-vivo-study
#15
Tomasz Przygodzki, Marcin Talar, Hassan Kassassir, Lukasz Mateuszuk, Jacek Musial, Cezary Watala
Numerous in vitro experiments have confirmed that a dysfunctional endothelium is characterized by, inter alia, a higher affinity for binding of platelets and leukocytes. However, there is still no direct evidence for greater interaction between platelets and intact endothelium in in vivo animal models of diabetes. Therefore, the present study examines the pro-adhesive properties of endothelium change in vivo as an effect of streptozotocin (STZ)-induced diabetes and the role of two key platelet receptors: GPIb-IX-V and GPIIb/IIIa...
July 26, 2017: Platelets
https://www.readbyqxmd.com/read/28733090/intravital-multiphoton-microscopy-as-a-tool-for-studying-renal-physiology-and-pathophysiology
#16
Ruben M Sandoval, Bruce A Molitoris
The kidney is a complex and dynamic organ with over 40 cell types, and tremendous structural and functional diversity. Intravital multi-photon microscopy, development of fluorescent probes and innovative software, have rapidly advanced the study of intracellular and intercellular processes within the kidney. Researchers can quantify the distribution, behavior, and dynamic interactions of up to four labeled chemical probes and proteins simultaneously and repeatedly in four dimensions (time), with subcellular resolution in near real time...
July 18, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28723288/science-to-practice-can-mr-imaging-cell-tracking-of-macrophage-infiltration-be-used-as-a-predictive-imaging-biomarker-for-transplanted-stem-cell-rejection
#17
Jeff W M Bulte
Ferumoxytol-enhanced magnetic resonance (MR) imaging of donor-matched and mismatched stem cell transplants demonstrated decreased signal intensity not only for a xenogeneic mismatch in species but, surprisingly, also for a syngeneic mismatch in sex. MR imaging findings were corroborated with intravital fluorescence microscopy (IVM), where nearly 90% of all ferumoxytol-containing cells were found to be macrophages. Hence, MR imaging cell tracking of infiltrating macrophages may have predictive value in determining whether transplanted stem cell rejection will occur...
August 2017: Radiology
https://www.readbyqxmd.com/read/28720647/in-situ-targeting-of-dendritic-cells-sets-tolerogenic-environment-and-ameliorates-cd4-t-cell-response-in-the-postischemic-liver
#18
Dominik Funken, Hellen Ishikawa-Ankerhold, Bernd Uhl, Maximilian Lerchenberger, Markus Rentsch, Doris Mayr, Steffen Massberg, Jens Werner, Andrej Khandoga
CD4(+) T cells recruited to the liver play a key role in the pathogenesis of ischemia/reperfusion (I/R) injury. The mechanism of their activation during alloantigen-independent I/R is not completely understood. We hypothesized that liver-resident dendritic cells (DCs) interact with CD4(+) T cells in the postischemic liver and that modulation of DCs or T-cell-DC interactions attenuates liver inflammation. In mice, warm hepatic I/R (90/120-240 min) was induced. Tolerogenic DCs were generated in situ by pretreatment of animals with the vitamin D analog paricalcitol...
July 18, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28720149/heparanase-promotes-neuroinflammatory-response-during-subarachnoid-hemorrhage-in-rats
#19
Benjarat Changyaleket, Zhao Zhong Chong, Randal O Dull, Danop Nanegrungsunk, Haoliang Xu
BACKGROUND: Heparanase, a mammalian endo-β-D-glucoronidase that specifically degrades heparan sulfate, has been implicated in inflammation and ischemic stroke. However, the role of heparanase in neuroinflammatory response in subarachnoid hemorrhage (SAH) has not yet been investigated. This study was designed to examine the association between heparanase expression and neuroinflammation during subarachnoid hemorrhage. METHODS: Rats were subjected to SAH by endovascular perforation, and the expression of heparanase was determined by Western blot analysis and immunofluorescence in the ipsilateral brain cortex at 24 h post-SAH...
July 18, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28715512/mouse-models-of-deep-vein-thrombosis
#20
T Schönfelder, S Jäckel, P Wenzel
The pathogenesis of venous thromboembolism (VTE) is still not completely understood. Experimental animals in which human deep vein thrombosis can be modeled are useful tools to investigate the pathogenesis of VTE. Besides the availability of transgenic and genetically modified mice, the use of high frequency ultrasound and intravital microscopy plays an important role in identifying thrombotic processes in mouse models. In this article, an overview about the application of various new technologies and existing mouse models is provided, and the impact of venous side branches on deep vein thrombosis in the mouse model is discussed...
2017: Gefässchirurgie: Zeitschrift Für Vaskuläre und Endovaskuläre Chirurgie
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