Impaired epidermal wound healing in vivo upon inhibition or deletion of Rac1.
To address the functions of Rac1 in keratinocytes of the basal epidermal layer and in the outer root sheath of hair follicles, we generated transgenic mice expressing a dominant inhibitory mutant of Rac, N17Rac1, under the control of the keratin 14 promoter. These mice do not exhibit an overt skin phenotype but show protracted skin wound re-epithelialization. Investigation into the underlying mechanisms revealed that in vivo both proliferation of wound-edge keratinocytes and centripetal migration of the neo-epidermis were impaired. Similar results were obtained in mice with an epidermis-specific deletion of Rac1. Primary epidermal keratinocytes that expressed the N17Rac1 transgene were less proliferative than control cells and showed reduced ERK1/2 phosphorylation upon growth factor stimulation. Adhesion, spreading, random migration and closure of scratch wounds in vitro were significantly inhibited on collagen I and, to a lesser extent, on fibronectin. Stroboscopic analysis of cell dynamics (SACED) of N17Rac1 transgenic and control keratinocytes identified decreased lamella-protrusion persistence in connection with increased ruffle frequency as a probable mechanism for the observed impairment of keratinocyte adhesion and migration. We conclude that Rac1 is functionally required for normal epidermal wound healing and, in this context, exerts a dual function - namely the regulation of keratinocyte proliferation and migration.
Tscharntke
Michael
M
Department of Dermatology, University of Cologne, Center for Molecular Medicine, (CMMC), Joseph-Stelzmann-Strasse 9, 50924 Cologne, Germany.
Pofahl
Ruth
R
Chrostek-Grashoff
Anna
A
Smyth
Neil
N
Niessen
Carien
C
Niemann
Catherin
C
Hartwig
Benedikt
B
Herzog
Volker
V
Klein
Helmut W
HW
Krieg
Thomas
T
Brakebusch
Cord
C
Haase
Ingo
I
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D004817:Epidermis / Q000503:physiopathology*; D015603:Keratinocytes / Q000166:cytology; D014945:Wound Healing; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000502:physiology*
C107953:RAC1 protein, human; D020830:rac1 GTP-Binding Protein