Name: biophysical assays to probe the mechanical properties of the interphase cell nucleus: substrate strain application and microneedle manipulation
Uploaded: 2011-09-14T12:00:00
Description: Watch this Scientific Journal Video about Biophysical Assays to Probe the Mechanical Properties of the Interphase Cell Nucleus: Substrate Strain Application and Microneedle Manipulation at JoVE.com

यह काम (HL082792 R01 और NS059348 R01) स्वास्थ्य और ब्रिघम और महिला अस्पताल कार्डियोवास्कुलर नेतृत्व समूह पुरस्कार के राष्ट्रीय संस्थानों द्वारा समर्थित किया गया.

1. सब्सट्रेट तनाव आवेदन सामान्यीकृत परमाणु तनाव की माप पारदर्शी, सेल संस्कृति की सतह के रूप में लोचदार सिलिकॉन झिल्ली के साथ तनाव व्यंजन की तैयारी भी शामिल है, व्यंजन पर चढ़ाना कोशिकाओं, और पह...

<ol>
	<li>Friedl, P., Wolf, K., Lammerding, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+mechanics+during+cell+migration.">Nuclear mechanics during cell migration.</a> Curr Opin Cell Biol. 23, 55-64 (2011).</li><li>Mejat, A., Misteli, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LINC+complexes+in+health+and+disease.">LINC complexes in health and disease.</a> Nucleus. 1, 40-52 (2010).</li><li>Worman, H. J., Fong, L. G., Muchir, A., Young, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laminopathies+and+the+long+strange+trip+from+basic+cell+biology+to+therapy.">Laminopathies and the long strange trip from basic cell biology to therapy.</a> J Clin Invest. 119, 1825-1836 (2009).</li><li>Caille, N., Tardy, Y., Meister, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+strain+field+in+endothelial+cells+subjected+to+uniaxial+deformation+of+their+substrate.">Assessment of strain field in endothelial cells subjected to uniaxial deformation of their substrate.</a> Ann Biomed Eng. 26, 409-416 (1998).</li><li>Lammerding, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lamins+A+and+C+but+not+lamin+B1+regulate+nuclear+mechanics.">Lamins A and C but not lamin B1 regulate nuclear mechanics.</a> J Biol Chem. 281, 25768-25780 (2006).</li><li>Lammerding, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abnormal+nuclear+shape+and+impaired+mechanotransduction+in+emerin-deficient+cells.">Abnormal nuclear shape and impaired mechanotransduction in emerin-deficient cells.</a> J Cell Biol. 170, 781-791 (2005).</li><li>Lammerding, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lamin+A/C+deficiency+causes+defective+nuclear+mechanics+and+mechanotransduction.">Lamin A/C deficiency causes defective nuclear mechanics and mechanotransduction.</a> J Clin Invest. 113, 370-378 (2004).</li><li>Verstraeten, V. L., Ji, J. Y., Cummings, K. S., Lee, R. T., Lammerding, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+mechanosensitivity+and+nuclear+stiffness+in+Hutchinson-Gilford+progeria+cells:+effects+of+farnesyltransferase+inhibitors.">Increased mechanosensitivity and nuclear stiffness in Hutchinson-Gilford progeria cells: effects of farnesyltransferase inhibitors.</a> Aging Cell. 7, 383-393 (2008).</li><li>Brooks, S. V., Zerba, E., Faulkner, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injury+to+muscle+fibres+after+single+stretches+of+passive+and+maximally+stimulated+muscles+in+mice.">Injury to muscle fibres after single stretches of passive and maximally stimulated muscles in mice.</a> J Physiol. 488, 459-469 (1995).</li><li>Maniotis, A. J., Chen, C. S., Ingber, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Demonstration+of+mechanical+connections+between+integrins,+cytoskeletal+filaments,+and+nucleoplasm+that+stabilize+nuclear+structure.">Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure.</a> Proc Natl Acad Sci. 94, 849-854 (1997).</li></ol>

सब्सट्रेट तनाव परख तनाव आवेदन सफलतापूर्वक किया गया है हमें और अन्य समूहों द्वारा उपयोग के लिए कोशिकाओं में प्रेरित परमाणु विरूपण यांत्रिक तनाव के अधीन अध्ययन और परमाणु कठोरता विशिष्ट परमा...

<table border="1"><tbody><tr><td> अभिकर्मक के नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> Fibronectin </td><td> Millipore </td><td> FC010 </td></tr><tr><td> MitoTracker रेड एफएम और ग्रीन एफएम </td><td> Invitrogen </td><td> M22425 और एम - +७५१४ </td></tr><tr><td> 33342 Hoechst </td><td> Invitrogen </td><td> H3570 </td></tr><tr><td> हांक buffered नमक खारा </td><td> Invitrogen </td><td> 14185 </td></tr><tr><td> Phenol मुक्त, DMEM </td><td> Invitrogen </td><td> 21063 </td></tr><tr><td> भ्रूण गोजातीय सीरम </td><td> Aleken बायोलॉजिकल </td><td> FBSS500 </td></tr><tr><td> पेनिसिलीन / स्ट्रेप्टोमाइसिन </td><td> सिग्मा </td><td> P0781-100ml </td></tr><tr><td> फिलामेंट के साथ Borosilicate ग्लास </td><td> Sutter साधन </td><td> BF100 78-10 </td></tr><tr><td> / चमक गैर प्रबलित सिलिकॉन कनवास, 0.005 चमक &quot; </td><td> विशेषता विनिर्माण इंक </td><td></td></tr><tr><td> Dulbecco फास्फेट खारा buffered </td><td> Invitrogen </td><td> 14200 </td></tr><tr><td> 35 मिमी गिलास नीचे संस्कृति व्यंजन (FluoroDish) </td><td> विश्व परिशुद्धता उपकरण, कांग्रेस </td><td> FD35-100 </td></tr><tr><td> Braycote 804 वैक्यूम तेल </td><td> Spi आपूर्ति </td><td> 05133A अटल बिहारी </td></tr></tbody></table>

biophysical assays to probe the mechanical properties of the interphase cell nucleus: substrate strain application and microneedle manipulation

In most eukaryotic cells, the nucleus is the largest organelle and is typically 2 to 10 times stiffer than the surrounding cytoskeleton; consequently, the physical properties of the nucleus contribute significantly to the overall biomechanical behavior of cells under physiological and pathological conditions. For example, in migrating neutrophils and invading cancer cells, nuclear stiffness can pose a major obstacle during extravasation or passage through narrow spaces within tissues.1 On the other hand, the nucleus of cells in mechanically active tissue such as muscle requires sufficient structural support to withstand repetitive mechanical stress. Importantly, the nucleus is tightly integrated into the cellular architecture; it is physically connected to the surrounding cytoskeleton, which is a critical requirement for the intracellular movement and positioning of the nucleus, for example, in polarized cells, synaptic nuclei at neuromuscular junctions, or in migrating cells.2 Not surprisingly, mutations in nuclear envelope proteins such as lamins and nesprins, which play a critical role in determining nuclear stiffness and nucleo-cytoskeletal coupling, have been shown recently to result in a number of human diseases, including Emery-Dreifuss muscular dystrophy, limb-girdle muscular dystrophy, and dilated cardiomyopathy.3 To investigate the biophysical function of diverse nuclear envelope proteins and the effect of specific mutations, we have developed experimental methods to study the physical properties of the nucleus in single, living cells subjected to global or localized mechanical perturbation. Measuring induced nuclear deformations in response to precisely applied substrate strain application yields important information on the deformability of the nucleus and allows quantitative comparison between different mutations or cell lines deficient for specific nuclear envelope proteins. Localized cytoskeletal strain application with a microneedle is used to complement this assay and can yield additional information on intracellular force transmission between the nucleus and the cytoskeleton. Studying nuclear mechanics in intact living cells preserves the normal intracellular architecture and avoids potential artifacts that can arise when working with isolated nuclei. Furthermore, substrate strain application presents a good model for the physiological stress experienced by cells in muscle or other tissues (e.g., vascular smooth muscle cells exposed to vessel strain). Lastly, while these tools have been developed primarily to study nuclear mechanics, they can also be applied to investigate the function of cytoskeletal proteins and mechanotransduction signaling.

Watch this Scientific Journal Video about Biophysical Assays to Probe the Mechanical Properties of the Interphase Cell Nucleus: Substrate Strain Application and Microneedle Manipulation at JoVE.com

Biophysical Assays to Probe the Mechanical Properties of the Interphase Cell Nucleus: Substrate Strain Application and Microneedle Manipulation

हम दो स्वतंत्र, खुर्दबीन आधारित उपकरण का वर्तमान वैश्विक या स्थानीय तनाव आवेदन के जवाब में एकल, रहने वाले अनुयायी कोशिकाओं में प्रेरित परमाणु और cytoskeletal विरूपण को मापने के लिए. इन तकनीकों के परमाणु कठोरता (यानी, विरूपता) का निर्धारण और नाभिक और cytoskeleton के बीच intracellular बल संचरण की जांच करने के लिए उपयोग किया जाता है.

Biophysical Assays अंतराप्रावस्था सेल नाभिक के यांत्रिक गुण जांच: सब्सट्रेट तनाव अनुप्रयोग और Microneedle हेरफेर

In most eukaryotic cells, the nucleus is the largest organelle and is typically 2 to 10 times stiffer than the surrounding cytoskeleton; consequently, the ...

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