Name: quantitative analysis of chromatin proteomes in disease
Uploaded: 2012-12-28T12:00:00
Description: Watch this Scientific Journal Video about Quantitative Analysis of Chromatin Proteomes in Disease at JoVE.com

प्रयोगशाला Vondriska NIH और UCLA में Laubisch बंदोबस्ती के राष्ट्रीय हृदय, फेफड़े और रक्त संस्थान से अनुदान द्वारा समर्थित है. EM जेनिफर एस UCLA में फिजियोलॉजी में Buchwald ग्रेजुएट फैलोशिप के प्राप्तकर्ता है, कोर्ट एक अमेरिकन हार्ट एसोसिएशन के पूर्व डॉक्टरेट फैलोशिप के प्राप्तकर्ता है, सांसद एक NIH रुथ Kirschstein पोस्ट डॉक्टरेट फैलोशिप के प्राप्तकर्ता है, और एस एफ के प्राप्तकर्ता है एक NIH K99 पुरस्कार से सम्मानित किया गया.

प्रयोगात्मक कार्यप्रवाह सात प्रमुख कदम (1 चित्रा) शामिल हैं. किसी भी नमूने है कि मास स्पेक्ट्रोमीटर पर चला जाएगा जुड़े काम के लिए एक प्रयोगशाला कोट, दस्ताने और बाल शुद्ध experimenter पहनते हैं और धूल और क?...

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पहले परमाणु अलगाव के लिए दो मुख्य तरीकों की समीक्षा की गई है: एक 27 Behrens एक गैर जलीय विलायक में lyophilized ऊतक homogenizing की तकनीक और दूसरा, एक संशोधन जिनमें से हम यहाँ का उपयोग एक जलीय sucrose / नमक के घोल में ऊतक homogenizing की ह?...

<table border="1"><tbody><tr><td> अभिकर्मक के नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> Dulbeco संशोधित ईगल मध्यम </td><td> Invitrogen </td><td> 11965 </td></tr><tr><td> Protease गोली </td><td> Roche </td><td> 04 159 001 693 </td></tr><tr><td> 100 सुक्ष्ममापी झरनी </td><td> BD फाल्कन </td><td> 352360 </td></tr><tr><td> Ultracut ultramicrotome </td><td> रीचर्ट </td><td></td></tr><tr><td> 100CX ट्रांसमिशन इलेक्ट्रॉन माइक्रोस्कोप </td><td> JEOL अमरीका, इंक, </td><td></td></tr><tr><td> ओरियल </td><td> BioRad </td><td> 161-0496 </td></tr><tr><td> Histone H2A एंटीबॉडी </td><td> सांता क्रुज़ </td><td> सुप्रीम कोर्ट-8648 </td></tr><tr><td> Nucleoporin p62 एंटीबॉडी </td><td> बी.डी. बायोसाइंसेज </td><td> 610498 </td></tr><tr><td> Adenine Nucleoज्वार ट्रांसपोर्टर एंटीबॉडी </td><td> सांता क्रुज़ </td><td> सुप्रीम कोर्ट-9299 </td></tr><tr><td> बीप प्रतिरक्षी </td><td> सांता क्रुज़ </td><td> SC-1050 </td></tr><tr><td> ट्यूबिलिन एंटीबॉडी </td><td> सिग्मा </td><td> T1568 </td></tr><tr><td> Histone H3 एंटीबॉडी </td><td> Abcam </td><td> ab1791 </td></tr><tr><td> Fibrillarin एंटीबॉडी </td><td> सेल संकेतन </td><td> C12C3 </td></tr><tr><td> SNRP70 एंटीबॉडी </td><td> Abcam </td><td> ab51266 </td></tr><tr><td> E2F-1 एंटीबॉडी </td><td> थर्मो फिशर </td><td> MS-879 </td></tr><tr><td> Retinoblastoma एंटीबॉडी </td><td> बी.डी. बायोसाइंसेज </td><td> 554136 </td></tr><tr><td> Hypoxia inducible कारक 1 एंटीबॉडी </td><td> Novus बायोलॉजिकल </td><td> NB100-469 </td></tr><tr><td> बीसीए प्रोटीन परख </td><td> थर्मो वैज्ञानिक </td><td> 23227 </td></tr><tr><td> रिवर्स चरण स्तंभ </td><td> नई Objective </td><td> PFC7515-B14-10 </td></tr><tr><td> BioWorks ब्राउज़र </td><td> थर्मो वैज्ञानिक </td><td></td></tr></tbody></table>

quantitative analysis of chromatin proteomes in disease

In the nucleus reside the proteomes whose functions are most intimately linked with gene regulation. Adult mammalian cardiomyocyte nuclei are unique due to the high percentage of binucleated cells,1 the predominantly heterochromatic state of the DNA, and the non-dividing nature of the cardiomyocyte which renders adult nuclei in a permanent state of interphase.2 Transcriptional regulation during development and disease have been well studied in this organ,3-5 but what remains relatively unexplored is the role played by the nuclear proteins responsible for DNA packaging and expression, and how these proteins control changes in transcriptional programs that occur during disease.6 In the developed world, heart disease is the number one cause of mortality for both men and women.7 Insight on how nuclear proteins cooperate to regulate the progression of this disease is critical for advancing the current treatment options.
Mass spectrometry is the ideal tool for addressing these questions as it allows for an unbiased annotation of the nuclear proteome and relative quantification for how the abundance of these proteins changes with disease. While there have been several proteomic studies for mammalian nuclear protein complexes,8-13 until recently14 there has been only one study examining the cardiac nuclear proteome, and it considered the entire nucleus, rather than exploring the proteome at the level of nuclear sub compartments.15 In large part, this shortage of work is due to the difficulty of isolating cardiac nuclei. Cardiac nuclei occur within a rigid and dense actin-myosin apparatus to which they are connected via multiple extensions from the endoplasmic reticulum, to the extent that myocyte contraction alters their overall shape.16 Additionally, cardiomyocytes are 40% mitochondria by volume17 which necessitates enrichment of the nucleus apart from the other organelles. Here we describe a protocol for cardiac nuclear enrichment and further fractionation into biologically-relevant compartments. Furthermore, we detail methods for label-free quantitative mass spectrometric dissection of these fractions-techniques amenable to in vivo experimentation in various animal models and organ systems where metabolic labeling is not feasible.

चित्रा 4 रिश्तेदार मात्रा का ठहराव के लिए इस प्रपत्र की उपयोगिता पर प्रकाश डाला गया. बाएं पैनल में दिखाया गया व्यक्ति monoisotopic पेप्टाइड अलग चूहों से मढ़ा चोटियों, है जो प्रोटीन HMGB1 डेटाबेस खोज के माध्?...

Watch this Scientific Journal Video about Quantitative Analysis of Chromatin Proteomes in Disease at JoVE.com

Quantitative Analysis of Chromatin Proteomes in Disease

मास स्पेक्ट्रोमेट्री अग्रिम में प्रोटीन अभिव्यक्ति और ऊतकों की एक मेजबान में संशोधन के उच्च throughput विश्लेषण की अनुमति दी है. Subcellular fractionation और रोग मॉडल, मात्रात्मक जन स्पेक्ट्रोमेट्री और जैव सूचना विज्ञान जैविक प्रणालियों में नए गुण प्रकट कर सकते हैं के साथ संयुक्त है. यहाँ वर्णित विधि हृदय रोग की सेटिंग में chromatin जुड़े प्रोटीन का विश्लेषण करता है और आसानी से दूसरे के लिए लागू है<em&gt; Vivo में</em&gt; मानव रोग के मॉडल.

रोग में Chromatin Proteomes के मात्रात्मक विश्लेषण

In the nucleus reside the proteomes whose functions are most intimately linked with gene regulation. Adult mammalian cardiomyocyte nuclei are unique due ...

Research

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Medicine

Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model

कैंसर मेटास्टेसिस के मात्रात्मक विश्लेषण एक एवियन भ्रूण मॉडल का उपयोग

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex ...

A Simple Method of Mouse Lung Intubation

माउस फेफड़े इंटुबैषेण का एक सरल तरीका

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Murine महाधमनी साइनस में atherosclerotic घावों की मात्रात्मक विश्लेषण और विशेषता

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Quantitative Magnetic Resonance Imaging of Skeletal Muscle Disease

कंकाल की मांसपेशी रोग के मात्रात्मक चुंबकीय अनुनाद इमेजिंग

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of ...

Quantitative Mass Spectrometric Profiling of Cancer-cell Proteomes Derived From Liquid and Solid Tumors

तरल और ठोस ट्यूमर से निकाली गई कैंसर सेल Proteomes के मात्रात्मक मास Spectrometric रूपरेखा

In-depth analyses of cancer cell proteomes are needed to elucidate oncogenic pathomechanisms, as well as to identify potential drug targets and diagnostic ...

Quantitative 3D In Silico Modeling (q3DISM) of Cerebral Amyloid-beta Phagocytosis in Rodent Models of Alzheimer's Disease

Quantitative 3D In Silico Modeling q3DISM of Cerebral Amyloid-beta Phagocytosis in Rodent Models of Alzheimer's Disease

मात्रात्मक 3 डी<em&gt; सिलिको</em&gt; मॉडलिंग अल्जाइमर रोग के कृंतक मॉडल में मस्तिष्क amyloid बीटा phagocytosis की (q3DISM)

Neuroinflammation is now recognized as a major etiological factor in neurodegenerative disease. Mononuclear phagocytes are innate immune cells responsible ...

Application of Granger Causality Analysis of the Directed Functional Connection in Alzheimer's Disease and Mild Cognitive Impairment

ग्रेंजर भिन्न विश्लेषण के निर्देश कार्यात्मक कनेक्शन में अल्जाइमर रोग और हल्के संज्ञानात्मक हानि के आवेदन

Impaired functional connectivity in the Default Mode Network (DMN) may be involved in the progression of Alzheimer&#39;s Disease (AD). The Posterior ...

In Vivo Morphometric Analysis of Human Cranial Nerves Using Magnetic Resonance Imaging in Meni&#232;re's Disease Ears and Normal Hearing Ears

In Vivo Morphometric Analysis of Human Cranial Nerves Using Magnetic Resonance Imaging in MeniÃƒÆ’Ã†'Ãƒâ€šÃ‚Â¨re's Disease Ears and Normal Hearing Ears

Vivo में मानव कपाल नसों के Morphometric विश्लेषण Menière's रोग कान और सामांय सुनने के कान में चुंबकीय अनुनाद इमेजिंग का उपयोग

Analysis of neural structures in Meni&#232;re's Disease (MD) is of importance, since a loss of such structures has previously been proposed for this ...

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&#8212;Application in Premanifest Huntington's Disease

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRIÃƒÆ’Ã‚Â¢ÃƒÂ¢Ã¢â‚¬Å¡Ã‚Â¬ÃƒÂ¢Ã¢â€šÂ¬Ã‚ÂApplication in Premanifest Huntington's Disease

पूरे मस्तिष्क विभाजन और संरचनात्मक मस्तिष्क एमआरआई के परिवर्तन बिंदु विश्लेषण-प्रकट Huntington की बीमारी में आवेदन

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy ...

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of ÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â²-aminopropionitrile-induced Aortic Aneurysm and Dissection

मात्रात्मक माइक्रो-सीटी विश्लेषण के एक माउस मॉडल में Aortopathy β-aminopropionitrile-प्रेरित महाधमनी धमनीविस्फार और विच्छेदन

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...