biostudies-literatureKim CSNational Institute of General Medical SciencesDivision of Chemical, Bioengineering, Environmental, and Transport SystemsNIGMS NIH HHSe10138https://www.ebi.ac.uk/biostudies/studies/S-EPMC6764804biostudies-literatureUnknown4(3)Re-epithelialization is a critical step in wound healing and results from the collective migration of keratinocytes. Previous work demonstrated that immobilized, but not soluble, epidermal growth factor (EGF) resulted in leader cell-specific activation of phospholipase C gamma 1 (PLCγ1) in HaCaT keratinocytes, and that this PLCγ1 activation was necessary to drive persistent cell migration. To determine the mechanism responsible for wound edge-localized PLCγ1 activation, we examined differences in cell area, cell-cell interactions, and EGF receptor (EGFR) localization between wound edge and bulk cells treated with vehicle, soluble EGF, or immobilized EGF. Our results support a multistep mechanism where EGFR translocation from the lateral membrane to the basolateral/basal membrane allows clustering in response to immobilized EGF. This analysis of factors regulating PLCγ1 activation is a crucial step toward developing therapies or wound dressings capable of modulating this signal and, consequently, cell migration.Bioengineering & translational medicineLeader cell PLCγ1 activation during keratinocyte collective migration is induced by EGFR localization and clustering.PMC6764804CBET‐1401584R01GM099031R01 GM099031Yang XKim CSMasters KSKreeger PKJacobsen SfalseLeader cell PLCγ1 activation during keratinocyte collective migration is induced by EGFR localization and clustering.Re-epithelialization is a critical step in wound healing and results from the collective migration of keratinocytes. Previous work demonstrated that immobilized, but not soluble, epidermal growth factor (EGF) resulted in leader cell-specific activation of phospholipase C gamma 1 (PLCγ1) in HaCaT keratinocytes, and that this PLCγ1 activation was necessary to drive persistent cell migration. To determine the mechanism responsible for wound edge-localized PLCγ1 activation, we examined differences in cell area, cell-cell interactions, and EGF receptor (EGFR) localization between wound edge and bulk cells treated with vehicle, soluble EGF, or immobilized EGF. Our results support a multistep mechanism where EGFR translocation from the lateral membrane to the basolateral/basal membrane allows clustering in response to immobilized EGF. This analysis of factors regulating PLCγ1 activation is a crucial step toward developing therapies or wound dressings capable of modulating this signal and, consequently, cell migration.2019-01-01T00:00:00Z2019 Sep2025-04-21T23:23:36.989Z2019-10-11T07:12:13ZS-EPMC67648043157279610.1002/btm2.10138