Determining the mechanical properties of plectin in mouse myoblasts and keratinocytes

Navid Bonakdar, Achim Schilling, Marina Spörrer, Pablo Lennert, Astrid Mainka, Lilli Winter, Gernot Walko, Gerhard Wiche, Ben Fabry, Wolfgang H Goldmann

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Plectin is the prototype of an intermediate filament (IF)-based cytolinker protein. It affects cells mechanically by interlinking and anchoring cytoskeletal filaments and acts as scaffolding and docking platform for signaling proteins to control cytoskeleton dynamics. The most common disease caused by mutations in the human plectin gene, epidermolysis bullosa simplex with muscular dystrophy (EBS-MD), is characterized by severe skin blistering and progressive muscular dystrophy. Therefore, we compared the biomechanical properties and the response to mechanical stress of murine plectin-deficient myoblasts and keratinocytes with wild-type cells. Using a cell stretching device, plectin-deficient myoblasts exhibited lower mechanical vulnerability upon external stress compared to wild-type cells, which we attributed to lower cellular pre-stress. Contrary to myoblasts, wild-type and plectin-deficient keratinocytes showed no significant differences. In magnetic tweezer measurements using fibronectin-coated paramagnetic beads, the stiffness of keratinocytes was higher than of myoblasts. Interestingly, cell stiffness, adhesion strength, and cytoskeletal dynamics were strikingly altered in plectin-deficient compared to wild-type myoblasts, whereas smaller differences were observed between plectin-deficient and wild-type keratinocytes, indicating that plectin might be more important for stabilizing cytoskeletal structures in myoblasts than in keratinocytes. Traction forces strongly correlated with the stiffness of plectin-deficient and wild-type myoblasts and keratinocytes. Contrary to that cell motility was comparable in plectin-deficient and wild-type myoblasts, but was significantly increased in plectin-deficient compared to wild-type keratinocytes. Thus, we postulate that the lack of plectin has divergent implications on biomechanical properties depending on the respective cell type.

Original languageEnglish
Pages (from-to)331-337
Number of pages7
JournalExperimental Cell Research
Volume331
Issue number2
DOIs
Publication statusPublished - 15 Feb 2015

Keywords

  • Animals
  • Biomechanical Phenomena
  • Cell Adhesion/genetics
  • Cell Line
  • Cell Movement
  • Keratinocytes/physiology
  • Magnetics
  • Mice
  • Myoblasts/physiology
  • Plectin/genetics
  • Stress, Mechanical
  • Stress, Physiological/genetics

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