TRPV4 Stimulation Level Regulates Ca2+-Dependent Control of Human Corneal Endothelial Cell Viability and Survival

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

Abstract

The functional contribution of transient receptor potential vanilloid 4 (TRPV4) expression in maintaining human corneal endothelial cells (HCEC) homeostasis is unclear. Accordingly, we determined the effects of TRPV4 gene and protein overexpression on responses modulating the viability and survival of HCEC. Q-PCR, Western blot, FACS analyses and fluorescence single-cell calcium imaging confirmed TRPV4 gene and protein overexpression in lentivirally transduced 12V4 cells derived from their parent HCEC-12 line. Although TRPV4 overexpression did not alter the baseline transendothelial electrical resistance (TEER), its cellular capacitance (Ccl) was larger than that in its parent. Scanning electron microscopy revealed that only the 12V4 cells developed densely packed villus-like protrusions. Stimulation of TRPV4 activity with GSK1016790A (GSK101, 10 µmol/L) induced larger Ca2+ transients in the 12V4 cells than those in the parental HCEC-12. One to ten nmol/L GSK101 decreased 12V4 viability, increased cell death rates and reduced the TEER, whereas 1 µmol/L GSK101 was required to induce similar effects in the HCEC-12. However, the TRPV4 channel blocker RN1734 (1 to 30 µmol/L) failed to alter HCEC-12 and 12V4 morphology, cell viability and metabolic activity. Taken together, TRPV4 overexpression altered both the HCEC morphology and markedly lowered the GSK101 dosages required to stimulate its channel activity.

Details

OriginalspracheEnglisch
Aufsatznummer281
Seitenumfang19
FachzeitschriftMembranes
Jahrgang12
Ausgabenummer3
PublikationsstatusVeröffentlicht - März 2022
Peer-Review-StatusJa

Externe IDs

Mendeley 20c8bf28-01cd-30e3-b47c-a188816ae6b3
ORCID /0000-0001-5624-1717/work/142239044
ORCID /0000-0003-3205-1876/work/142256468
ORCID /0000-0002-0320-4223/work/150884947

Schlagworte

Fächergruppen, Lehr- und Forschungsbereiche, Fachgebiete nach Destatis

Schlagwörter

  • Cell surface differentiation, Cell survival, Human corneal endothelial cells, Intracellular Ca signaling, Transepithelial electrical resistance, Transient receptor potential vanilloid subtype 4, Intracellular Ca2+ signaling

Bibliotheksschlagworte