Universität zu Köln

Jahnel, Oliver, Hoffmann, Bernd ORCID: 0000-0002-3803-8835, Merkel, Rudolf ORCID: 0000-0003-3178-3282, Bossinger, Olaf and Leube, Rudolf E. (2016). Mechanical Probing of the Intermediate Filament-Rich Caenorhabditis Elegans Intestine. In: Methods in Enzymology, S. 681 - 707. SAN DIEGO: ELSEVIER ACADEMIC PRESS INC. ISBN 978-0-12-803491-0; 978-0-12-803470-5

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Abstract

It is commonly accepted that intermediate filaments have an important mechanical function. This function relies not only on intrinsic material properties but is also determined by dynamic interactions with other cytoskeletal filament systems, distinct cell adhesion sites, and cellular organelles which are fine-tuned by multiple signaling pathways. While aspects of these properties and processes can be studied in vitro, their full complexity can only be understood in a viable tissue context. Yet, suitable and easily accessible model systems for monitoring tissue mechanics at high precision are rare. We show that the dissected intestine of the genetic model organism Caenorhabditis elegans fulfills this requirement. The 20 intestinal cells, which are arranged in an invariant fashion, are characterized by a dense subapical mesh of intermediate filaments that are attached to the C. elegans apical junction. We present procedures to visualize details of the characteristic intermediate filament-junctional complex arrangement in living animals. We then report on methods to prepare intestines with a fully intact intermediate filament cytoskeleton and detail procedures to assess their viability. A dual micropipette assay is described to measure mechanical properties of the dissected intestine while monitoring the spatial arrangement of the intermediate filament system. Advantages of this approach are (i) the high reproducibility of measurements because of the uniform architecture of the intestine and (ii) the high degree of accessibility allowing not only mechanical manipulation of an intact tissue but also control of culture medium composition and addition of drugs as well as visualization of cell structures. With this method, examination of worms carrying mutations in the intermediate filament system, its interacting partners and its regulators will become feasible.

Item Type:	Book Section, Proceedings Item or annotation in a legal commentary
Creators:	Creators Email ORCID ORCID Put Code Jahnel, Oliver UNSPECIFIED UNSPECIFIED UNSPECIFIED Hoffmann, Bernd UNSPECIFIED orcid.org/0000-0002-3803-8835 UNSPECIFIED Merkel, Rudolf UNSPECIFIED orcid.org/0000-0003-3178-3282 UNSPECIFIED Bossinger, Olaf UNSPECIFIED UNSPECIFIED UNSPECIFIED Leube, Rudolf E. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-292786
DOI:	10.1016/bs.mie.2015.08.030
Title of Book:	Methods in Enzymology
Series Name:	Methods Enzymol.
Volume:	568
Page Range:	S. 681 - 707
Date:	2016
Publisher:	ELSEVIER ACADEMIC PRESS INC
Place of Publication:	SAN DIEGO
ISSN:	0076-6879
ISBN:	978-0-12-803491-0; 978-0-12-803470-5
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language VISCOELASTIC PROPERTIES; ESSENTIAL ROLES; CELL STIFFNESS; TERMINAL WEB; KERATINS; NETWORK; ORGANOGENESIS; FLUORESCENCE; ORGANIZATION; INTEGRITY Multiple languages Biochemical Research Methods; Biochemistry & Molecular Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/29278