Ligand-induced dynamic membrane changes and cell deletion conferred by vanilloid receptor 1

J Biol Chem. 2001 Apr 6;276(14):11021-30. doi: 10.1074/jbc.M008392200. Epub 2000 Dec 21.

Abstract

The real time dynamics of vanilloid-induced cytotoxicity and the specific deletion of nociceptive neurons expressing the wild-type vanilloid receptor (VR1) were investigated. VR1 was C-terminally tagged with either the 27-kDa enhanced green fluorescent protein (eGFP) or a 12-amino acid epsilon-epitope. Upon exposure to resiniferatoxin, VR1eGFP- or VR1epsilon-expressing cells exhibited pharmacological responses similar to those of cells expressing the untagged VR1. Within seconds of vanilloid exposure, the intracellular free calcium ([Ca(2+)](i)) was elevated in cells expressing VR1. A functional pool of VR1 also was localized to the endoplasmic reticulum that, in the absence of extracellular calcium, also was capable of releasing calcium upon agonist treatment. Confocal imaging disclosed that resiniferatoxin treatment induced vesiculation of the mitochondria and the endoplasmic reticulum ( approximately 1 min), nuclear membrane disruption (5-10 min), and cell lysis (1-2 h). Nociceptive primary sensory neurons endogenously express VR1, and resiniferatoxin treatment induced a sudden increase in [Ca(2+)](i) and mitochondrial disruption which was cell-selective, as glia and non-VR1-expressing neurons were unaffected. Early hallmarks of cytotoxicity were followed by specific deletion of VR1-expressing cells. These data demonstrate that vanilloids disrupt vital organelles within the cell body and, if administered to sensory ganglia, may be employed to rapidly and selectively delete nociceptive neurons.

MeSH terms

  • Animals
  • Biological Transport
  • COS Cells
  • Capsaicin / pharmacology
  • Green Fluorescent Proteins
  • Ligands
  • Luminescent Proteins
  • Microscopy, Confocal
  • Receptors, Drug / agonists
  • Receptors, Drug / metabolism*
  • Signal Transduction*
  • TRPV Cation Channels

Substances

  • Ligands
  • Luminescent Proteins
  • Receptors, Drug
  • TRPV Cation Channels
  • TRPV1 receptor
  • Green Fluorescent Proteins
  • Capsaicin