Name: visualization of vascular ca2+ signaling triggered by paracrine derived ros
Uploaded: 2011-12-21T13:00:00
Description: Watch this Scientific Journal Video about Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS at JoVE.com

यह काम राष्ट्रीय संस्थानों स्वास्थ्य अनुदान के एम.एम. (HL086699 R01, HL086699 01A2S1, 1S10RR027327-01) द्वारा समर्थित किया गया. हमारे लेख आंशिक रूप से कार्ल Zeiss MicroImaging LLC द्वारा समर्थित है.

प्रतिक्रियाशील ऑक्सीजन प्रजातियों रहते ऑक्सीकरण संवेदनशील रंजक (1 और 2) का उपयोग कर या confocal माइक्रोस्कोपी द्वारा प्लाज्मिड सेंसर (3 एवं 4) का उपयोग कोशिकाओं में मापा जा सकता है है. 1. साइटोसोलिक ROS ?...

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विधि यहाँ वर्णित कोशिकाओं रहने वाले या तो ऑक्सीकरण संवेदनशील रंजक या प्लाज्मिड सेंसर का उपयोग करने में प्रतिक्रियाशील ऑक्सीजन प्रजातियों के तेजी से और मात्रात्मक माप की अनुमति देता है. TLRs के agonists (टोल ?...

<table border="1"><tbody><tr><td> अभिकर्मक </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> Attofluor सेल कक्ष </td><td> Invitrogen </td><td> A7816 </td></tr><tr><td> Antimycin एक </td><td> सिग्मा Aldrich </td><td> A8674 </td></tr><tr><td> DMEM कम ग्लूकोज मध्यम </td><td> Invitrogen </td><td> 10567-014 </td></tr><tr><td> Endothelial वृद्धि कारक पूरक (ECGs) </td><td> Upstate </td><td> 02-102 </td></tr><tr><td> भ्रूण गोजातीय सीरम </td><td> Invitrogen </td><td> 12662011 </td></tr><tr><td> G418 </td><td> Invitrogen </td><td> 10131-027 </td></tr><tr><td> जेलाटीन </td><td> सिग्मा Aldrich </td><td> G1393 </td></tr><tr><td> एच 2 DCFDA </td><td> Invitrogen </td><td> D-399 </td></tr><tr><td> LPS </td><td> सिग्मा Aldrich </td><td> L4516 </td></tr><tr><td> LTA </td><td> सिग्मा Aldrich </td><td> L2515 </td></tr><tr><td> MitoSOX लाल </td><td> Invitrogen </td><td> M36008 </td></tr><tr><td> Opti-सदस्य मैं सीरम मध्यम कम </td><td> Invitrogen </td><td> 51985091 </td></tr><tr><td> पेन / Strep (10x) </td><td> Invitrogen </td><td> 15140163 </td></tr><tr><td> pHyPer - cyto </td><td> Evrogen </td><td> FP941 </td></tr><tr><td> pHyPer - dMito </td><td> Evrogen </td><td> FP942 </td></tr><tr><td> पोलीफोनिक (मैं: सी) </td><td> सिग्मा Aldrich </td><td> P0913 </td></tr><tr><td> चश्मे सॉफ्टवेयर 5.0 </td><td> GraphPad सॉफ्टवेयर इंक, </td><td></td></tr><tr><td> SigmaPlot 11.0 </td><td> Systat सॉफ्टवेयर इंक, </td><td></td></tr><tr><td> Trypsin EDTA (10x) </td><td> Invitrogen </td><td> 15400054 </td></tr><tr><td> टी 25 बोतल </td><td> कोर्निंग </td><td> 430639 </td></tr><tr><td> टी 75 बोतल </td><td> बी.डी. फाल्कन </td><td> 353136 </td></tr><tr><td> टी 96 में अच्छी तरह सेसी अच्छी तरह से थाली माइक्रो </td><td> बी.डी. फाल्कन </td><td> 353072 </td></tr><tr><td> ज़ेन 2009 सॉफ्टवेयर </td><td> कार्ल Zeiss 510 मेटा confocal माइक्रोस्कोपी </td><td></td></tr></tbody></table>

visualization of vascular ca2+ signaling triggered by paracrine derived ros

Oxidative stress has been implicated in a number of pathologic conditions including ischemia/reperfusion damage and sepsis. The concept of oxidative stress refers to the aberrant formation of ROS (reactive oxygen species), which include O2&bull;-, H2O2, and hydroxyl radicals. Reactive oxygen species influences a multitude of cellular processes including signal transduction, cell proliferation and cell death1-6. ROS have the potential to damage vascular and organ cells directly, and can initiate secondary chemical reactions and genetic alterations that ultimately result in an amplification of the initial ROS-mediated tissue damage. A key component of the amplification cascade that exacerbates irreversible tissue damage is the recruitment and activation of circulating inflammatory cells. During inflammation, inflammatory cells produce cytokines such as tumor necrosis factor-&alpha; (TNF&alpha;) and IL-1 that activate endothelial cells (EC) and epithelial cells and further augment the inflammatory response7. Vascular endothelial dysfunction is an established feature of acute inflammation. Macrophages contribute to endothelial dysfunction during inflammation by mechanisms that remain unclear. Activation of macrophages results in the extracellular release of O2&bull;- and various pro-inflammatory cytokines, which triggers pathologic signaling in adjacent cells8. NADPH oxidases are the major and primary source of ROS in most of the cell types. Recently, it is shown by us and others9,10 that ROS produced by NADPH oxidases induce the mitochondrial ROS production during many pathophysiological conditions. Hence measuring the mitochondrial ROS production is equally important in addition to measuring cytosolic ROS. Macrophages produce ROS by the flavoprotein enzyme NADPH oxidase which plays a primary role in inflammation. Once activated, phagocytic NADPH oxidase produces copious amounts of O2&bull;- that are important in the host defense mechanism11,12. Although paracrine-derived O2&bull;- plays an important role in the pathogenesis of vascular diseases, visualization of paracrine ROS-induced intracellular signaling including Ca2+ mobilization is still hypothesis. We have developed a model in which activated macrophages are used as a source of O2&bull;- to transduce a signal to adjacent endothelial cells. Using this model we demonstrate that macrophage-derived O2&bull;- lead to calcium signaling in adjacent endothelial cells.

Watch this Scientific Journal Video about Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS at JoVE.com

Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS

Paracrine व्युत्पन्न endothelial सीए 2 + संकेत के ROS प्रेरण visualizing में अंतर्दृष्टि लाभ के लिए एक कुशल विधि वर्णित है. इस विधि paracrine व्युत्पन्न ROS मापने संवहनी endothelial कोशिकाओं में एक सह संस्कृति मॉडल में सीए 2 + जुटाना ट्रिगर का लाभ लेता है.

संवहनी Ca के विज़ुअलाइज़ेशन 2 + संकेतन Paracrine व्युत्पन्न ROS द्वारा ट्रिगर

Oxidative stress has been implicated in a number of pathologic conditions including ischemia/reperfusion damage and sepsis. The concept of oxidative ...

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Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS

Techniques for Imaging Ca2+ Signaling in Human Sperm

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Ca इमेजिंग तकनीक के लिए 2 + मानव शुक्राणु में संकेतन

Fluorescence microscopy of cells loaded with fluorescent, Ca2+-sensitive dyes is used for measurement of spatial and temporal aspects of Ca2+ signaling in ...

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डिसी परफ़्यूज़न में संवहनी विज़ुअलाइज़ेशन के लिए एक विधि के रूप में<em&gt; अंबिस्टोम मैक्सिकनम</em

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उच्च वोल्टेज क्रायो-इलेक्ट्रॉन टोमोग्राफी द्वारा Cyanobacteria में डीएनए संपीड़न के दृश्य

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रेटिना क्रायो-वर्गों, पूरे mounts, और Hypotonic Microvascular Pericytes के Immunohistochemical दृश्य के लिए अलग Vasculature तैयारी

Retinal pericytes play an important role in many diseases of the eye. Immunohistochemical staining techniques of retinal vessels and microvascular ...

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कोशिकाओं के बीच Paracrine सिगनल का अध्ययन करने के लिए एक समीपस्थ संस्कृति पद्धति

Intercellular interactions play an important role in many biological processes, including tumor progression, immune responses, angiogenesis, and ...

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Spatio-लौकिक मानचित्रण और कण विश्लेषण तकनीक के अनुप्रयोगों Intracellular Ca के लिए2 + सीटू में संकेतन

Ca2+ imaging of isolated cells or specific types of cells within intact tissues often reveals complex patterns of Ca2+ signaling. This activity requires ...