Loss of calpains-1 and -2 prevents repair of plasma membrane scrape injuries, but not small pores, and induces a severe muscular dystrophy

Am J Physiol Cell Physiol. 2020 Jun 1;318(6):C1226-C1237. doi: 10.1152/ajpcell.00408.2019. Epub 2020 Apr 29.

Abstract

The ubiquitous calpains, calpain-1 and -2, play important roles in Ca2+-dependent membrane repair. Mechanically active tissues like skeletal muscle are particularly reliant on mechanisms to repair and remodel membrane injury, such as those caused by eccentric damage. We demonstrate that calpain-1 and -2 are master effectors of Ca2+-dependent repair of mechanical plasma membrane scrape injuries, although they are dispensable for repair/removal of small wounds caused by pore-forming agents. Using CRISPR gene-edited human embryonic kidney 293 (HEK293) cell lines, we established that loss of both calpains-1 and -2 (CAPNS1-/-) virtually ablates Ca2+-dependent repair of mechanical scrape injuries but does not affect injury or recovery from perforation by streptolysin-O or saponin. In contrast, cells with targeted knockout of either calpain-1 (CAPN1-/-) or -2 (CAPN2-/-) show near-normal repair of mechanical injuries, inferring that both calpain-1 and calpain-2 are equally capable of conducting the cascade of proteolytic cleavage events to reseal a membrane injury, including that of the known membrane repair agent dysferlin. A severe muscular dystrophy in a murine model with skeletal muscle knockout of Capns1 highlights vital roles for calpain-1 and/or -2 for health and viability of skeletal muscles not compensated for by calpain-3 (CAPN3). We propose that the dystrophic phenotype relates to loss of maintenance of plasma membrane/cytoskeletal networks by calpains-1 and -2 in response to directed and dysfunctional Ca2+-signaling, pathways hyperstimulated in the context of membrane injury. With CAPN1 variants associated with spastic paraplegia, a severe dystrophy observed with muscle-specific loss of calpain-1 and -2 activity identifies CAPN2 and CAPNS1 as plausible candidate neuromuscular disease genes.

Keywords: Ca2+; calpain; cytoskeletal remodeling; membrane injury; membrane repair; muscular dystrophy; pore-forming toxins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / pharmacology
  • Calcium Signaling
  • Calpain / deficiency*
  • Calpain / genetics
  • Cell Membrane / drug effects
  • Cell Membrane / enzymology*
  • Cell Membrane / pathology
  • Disease Models, Animal
  • Dysferlin / deficiency
  • Dysferlin / genetics
  • Female
  • HEK293 Cells
  • Humans
  • Male
  • Mice, Knockout
  • Muscle, Skeletal / enzymology*
  • Muscle, Skeletal / pathology
  • Muscular Dystrophies, Limb-Girdle / enzymology*
  • Muscular Dystrophies, Limb-Girdle / genetics
  • Muscular Dystrophies, Limb-Girdle / pathology
  • Muscular Dystrophy, Animal / enzymology*
  • Muscular Dystrophy, Animal / genetics
  • Muscular Dystrophy, Animal / pathology
  • Saponins / pharmacology
  • Severity of Illness Index
  • Streptolysins / pharmacology

Substances

  • Bacterial Proteins
  • Dysf protein, mouse
  • Dysferlin
  • Saponins
  • Streptolysins
  • streptolysin O
  • Calpain
  • Capns1 protein, mouse
  • CAPN1 protein, human
  • CAPNS1 protein, human
  • Capn1 protein, mouse
  • CAPN2 protein, human
  • Capn2 protein, mouse