Funktionelle Charakterisierung von TRPV3 in kutaner Schmerzwahrnehmung
Zusammenfassung der Projektergebnisse
TRPV3, a member of the transient receptor potential (TRP) channel family, has been implicated as cellular sensor for thermal and chemical stimuli in the skin. A contribution of TRPV3 to cutaneous nociception has been suggested. Recent findings link TRPV3 mediated signaling to skin disease phenotypes supporting a primary function of the channel in skin homeostasis and suggesting that TRPV3 contribution to pain transduction may only be a consequence of such. Understanding its physiological function and potential contribution to cutaneous nociception requires knowledge about its activation/modulation and precise localization within the skin. A high-throughput screen designed to unravel endogenous activators of TRPV3 failed. Different strategies applied to acquire unambiguous expression data were inconclusive, but initiated a study on MscL, a bacterial mechanosensitive channel. MscL has one of the largest pores in nature and functions as a safety valve in bacteria releasing solutes during hypoosmotic downshock. The finding that MscL functionally expresses in mammalian cell membranes supported the idea to apply the channel as a tool for fast, timed and controllable uptake of molecules to cells. Charge-induced activation was adopted as a method to activate MscL, and fluorescently-labeled model cargo used to assess delivery efficacy of different sized molecules. Phalloidin, a cell-impermeable bi-cyclic peptide toxin from Amanita phalloides and specific marker for actin filaments, was successfully delivered supporting the use of MscL as a tool in cell biology for the delivery of bioactive molecules.
Projektbezogene Publikationen (Auswahl)
- IUPHAR Database, Transient Receptor Potential Channel: TRPV3
Julia F. Doerner & David E. Clapham
- (2012). Controlled delivery of bioactive molecules into live cells using the bacterial mechanosensitive channel MscL. Nature Commun 3, 990
Doerner JF, Febvay S, & Clapham DE
