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Functional Analysis of LINC Complexes in the Skin

  • Iakowos Karakesisoglou
  • Carmen Mroß
  • Angelika A. NoegelEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1840)

Abstract

The genome in eukaryotic cells is encased by two intricate and interconnected concentric membranes, which together with the underlying nuclear lamina form the nuclear envelope (NE). Two fundamental macromolecular structures are embedded within the nuclear envelope: the nuclear pore (NPC) and the LINC complex. The former perforates the nucleus controlling biomolecule trafficking between the nucleoplasm and the cytoplasm, while the latter integrates the nucleus via the cytoskeleton to the extracellular matrix. LINC complex structural and functional integrity is of utmost importance for various fundamental cellular functions. Mechanical forces are relayed into the nuclear interior via the LINC complex, which controls lamina organization, chromosome dynamics, and genome organization and stability. Thus, LINC constituents play pivotal roles in cellular architecture including organelle positioning, cell movement, tissue assembly, organ homeostasis, and organismal aging. The LINC complex oligomeric core contains several multi-isomeric, multifunctional, and often tissue-specific proteins. Therefore, for a proper functional analysis, genetic mouse models are an invaluable resource. Herein, we focus on the LINC complex roles in the skin and describe methods that enable the successful isolation of primary embryonic fibroblast and newborn skin cells, which can be then investigated functionally in vitro.

Key words

Nesprin SUN Mouse embryonic fibroblasts Primary keratinocytes Primary fibroblasts Directed cell migration 

Notes

Acknowledgments

This work was supported by Cologne Cluster of Excellence on Cellular Stress Responses in Aging-Associated Diseases (CECAD), the International Graduate School in Development Health and Disease (IGSDHD), and the BBSRC (Biotechnology and Biological Sciences Research Council).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Iakowos Karakesisoglou
    • 1
  • Carmen Mroß
    • 2
    • 3
  • Angelika A. Noegel
    • 2
    • 3
    Email author
  1. 1.Department of BiosciencesUniversity of DurhamDurhamUK
  2. 2.Institute of Biochemistry I, Medical FacultyUniversity Hospital CologneKölnGermany
  3. 3.Center for Molecular Medicine Cologne (CMMC) and Cologne Cluster of Excellence on Cellular Stress Responses in Aging-Associated Diseases (CECAD)University of CologneKölnGermany

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