The LINC Complex: Methods and Protocols

✍ Scribed by Gregg G. Gundersen, Howard J. Worman

Publisher: Springer New York;Humana Press
Year: 2018
Tongue: English
Leaves: 332
Series: Methods in Molecular Biology 1840
Edition: 1st ed.
Category: Library

No coin nor oath required. For personal study only.

✦ Synopsis

This volume describes research methodologies and approaches used to study the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex and its cellular functions. Chapters detail structural and biochemical analysis of LINC complexes, mechanical aspects of the LINC complex, analysis of the LINC complex in model systems and development, and LINC complex in mammalian tissue, organs, and disease. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, The LINC Complex: Methods and Protocols aims to ensure successful results in the further study of this vital field.

✦ Table of Contents

Front Matter ....Pages i-xiv
Front Matter ....Pages 1-1
EM Tomography of Meiotic LINC Complexes (Marie-Christin Spindler, Frederik Helmprobst, Christian Stigloher, Ricardo Benavente)....Pages 3-15
Recombinant Purification of the Periplasmic Portion of the LINC Complex (Victor E. Cruz, Thomas U. Schwartz)....Pages 17-23
Analysis of High Molecular Weight Isoforms of Nesprin-1 and Nesprin-2 with Vertical Agarose Gel Electrophoresis (Chloe Potter, Didier Hodzic)....Pages 25-33
Analysis of Nesprin-2 Interaction with Its Binding Partners and Actin (Susumu Antoku, Gregg G. Gundersen)....Pages 35-43
Interactions of Nesprin-4-Containing LINC Complexes in Outer Hair Cells Explored by BioID (Brian Burke)....Pages 45-56
Front Matter ....Pages 57-57
Using Nesprin Tension Sensors to Measure Force on the LINC Complex (Paul T. Arsenovic, Daniel E. Conway)....Pages 59-71
Analyzing Mechanotransduction Through the LINC Complex in Isolated Nuclei (Nejma Belaadi, Angélique Millon-Frémillon, Julien Aureille, Christophe Guilluy)....Pages 73-80
Direct Force Probe for Nuclear Mechanics (Vincent J. Tocco, Srujana Neelam, Qiao Zhang, Richard B. Dickinson, Tanmay P. Lele)....Pages 81-90
Centrifugal Displacement of Nuclei in Adherent Cells to Study LINC Complex-Dependent Mechanisms of Homeostatic Nuclear Positioning (Ruijun Zhu, Gregg G. Gundersen)....Pages 91-100
Assembly and Use of a Microfluidic Device to Study Cell Migration in Confined Environments (Jeremy Keys, Aaron Windsor, Jan Lammerding)....Pages 101-118
Front Matter ....Pages 119-119
Investigating LINC Complex Protein Homo-oligomerization in the Nuclear Envelopes of Living Cells Using Fluorescence Fluctuation Spectroscopy (Jared Hennen, Isaac Angert, Kwang-Ho Hur, G. W. Gant Luxton, Joachim D. Mueller)....Pages 121-135
Functional Analysis of the Yeast LINC Complex Using Fluctuation Spectroscopy and Super-Resolution Imaging (Jay R. Unruh, Brian D. Slaughter, Sue L. Jaspersen)....Pages 137-161
Genetic Analysis of Nuclear Migration and Anchorage to Study LINC Complexes During Development of Caenorhabditis elegans (Heidi N. Fridolfsson, Leslie A. Herrera, James N. Brandt, Natalie E. Cain, Greg J. Hermann, Daniel A. Starr)....Pages 163-180
High-Resolution Imaging Methods to Analyze LINC Complex Function During Drosophila Muscle Development (Alexander L. Auld, Mary Ann Collins, Torrey R. Mandigo, Eric S. Folker)....Pages 181-203
Computational Methods for Studying the Plant Nucleus (Axel Poulet, Xiao Zhou, Kentaro Tamura, Iris Meier, Christophe Tatout, Katja Graumann et al.)....Pages 205-219
Investigation of Nuclear Periphery Protein Interactions in Plants Using the Membrane Yeast Two-Hybrid (MbY2H) System (Maxime Voisin, Emmanuel Vanrobays, Christophe Tatout)....Pages 221-235
Immunolabeling Protocols for Studying Meiosis in Plant Mutants Defective for Nuclear Envelope Components (Javier Varas, Mónica Pradillo)....Pages 237-247
Front Matter ....Pages 249-249
Generation and Analysis of Striated Muscle Selective LINC Complex Protein Mutant Mice (Matthew J. Stroud, Xi Fang, Jennifer Veevers, Ju Chen)....Pages 251-281
An In Vitro System to Measure the Positioning, Stiffness, and Rupture of the Nucleus in Skeletal Muscle (William Roman, Mafalda R. Pimentel, Edgar R. Gomes)....Pages 283-293
Functional Analysis of LINC Complexes in the Skin (Iakowos Karakesisoglou, Carmen Mroß, Angelika A. Noegel)....Pages 295-306
Detection of SUN1 Splicing Variants at the mRNA and Protein Levels in Cancer (Ayaka Matsumoto, Nariaki Matsuura, Miki Hieda)....Pages 307-319
Next-Generation Sequencing and Mutational Analysis: Implications for Genes Encoding LINC Complex Proteins (Peter L. Nagy, Howard J. Worman)....Pages 321-336
Back Matter ....Pages 337-338

✦ Subjects

Life Sciences; Cell Biology

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