Spontaneous skin erosions and reduced skin and corneal wound healing characterize CLIC4(NULL) mice

Am J Pathol. 2012 Jul;181(1):74-84. doi: 10.1016/j.ajpath.2012.03.025. Epub 2012 May 18.


Cutaneous wound healing is a complex process involving blood clotting, inflammation, migration of keratinocytes, angiogenesis, and, ultimately, tissue remodeling and wound closure. Many of these processes involve transforming growth factor-β (TGF-β) signaling, and mice lacking components of the TGF-β signaling pathway are defective in wound healing. We show herein that CLIC4, an integral component of the TGF-β pathway, is highly up-regulated in skin wounds. We genetically deleted murine CLIC4 and generated a colony on a C57Bl/6 background. CLIC4(NULL) mice were viable and fertile but had smaller litters than did wild-type mice. After 6 months of age, up to 40% of null mice developed spontaneous skin erosions. Reepithelialization of induced full-thickness skin wounds and superficial corneal wounds was delayed in CLIC4(NULL) mice, resolution of inflammation was delayed, and expression of β4 integrin and p21 was reduced in lysates of constitutive and wounded CLIC4(NULL) skin. The induced level of phosphorylated Smad2 in response to TGF-β was reduced in cultured CLIC4(NULL) keratinocytes relative to in wild-type cells, and CLIC4(NULL) keratinocytes migrated slower than did wild-type keratinocytes and did not increase migration in response to TGF-β. CLIC4(NULL) keratinocytes were also less adherent on plates coated with matrix secreted by wild-type keratinocytes. These results indicate that CLIC4 participates in skin healing and corneal wound reepithelialization through enhancement of epithelial migration by a mechanism that may involve a compromised TGF-β pathway.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Animals
  • Cell Adhesion / physiology
  • Cell Movement / physiology
  • Cells, Cultured
  • Chloride Channels / deficiency
  • Chloride Channels / physiology*
  • Cornea / pathology
  • Cornea / physiology
  • Corneal Injuries*
  • Dose-Response Relationship, Drug
  • Keratinocytes / drug effects
  • Keratinocytes / metabolism
  • Keratinocytes / physiology
  • Mice
  • Mice, Knockout
  • Microscopy, Confocal
  • Mitochondrial Proteins / deficiency
  • Mitochondrial Proteins / physiology*
  • Proteins / metabolism
  • Signal Transduction / physiology
  • Skin / injuries*
  • Skin / metabolism
  • Skin / pathology
  • Skin Ulcer / pathology
  • Skin Ulcer / physiopathology*
  • Time Factors
  • Transforming Growth Factor beta / administration & dosage
  • Transforming Growth Factor beta / pharmacology
  • Wound Healing / physiology*


  • CLIC protein, mouse
  • Chloride Channels
  • Mitochondrial Proteins
  • Proteins
  • Transforming Growth Factor beta