Beta-bungarotoxin is a potent inducer of apoptosis in cultured rat neurons by receptor-mediated internalization

Eur J Neurosci. 2001 Sep;14(5):821-8. doi: 10.1046/j.0953-816x.2001.01699.x.

Abstract

The neurotoxic phospholipase A(2), beta-bungarotoxin (beta-BuTx), is a component of the snake venom from the Taiwanese banded krait Bungarus multicinctus. beta-BuTx affects presynaptic nerve terminal function of the neuromuscular junction and induces widespread neuronal cell death throughout the mammalian and avian CNS. To analyse the initial events of beta-BuTx-mediated cell death, the toxin was applied to cultured rat hippocampal neurons where it induced neuronal cell death in a concentration-dependent manner (EC(50) approximately equal to 5 x 10(-13) M) within 24 h. Fluorescence labelled beta-BuTx was completely incorporated by neurons within < 10 min. Binding and uptake of beta-BuTx, as well as induction of cell death, were efficiently antagonized by preincubation with dendrotoxin I, a blocker of voltage-gated potassium channels devoid of phospholipase activity. Binding of beta-BuTx was selective for neurofilament-positive cells. As evident from intense annexin-V and TUNEL stainings, application of beta-BuTx induced apoptotic cell death exclusively in neurons, leaving astrocytes unaffected. No evidence was obtained for any contribution of either caspases or calpains to beta-BuTx-induced apoptosis, consistent with the inability of the inhibitors Z-Asp-DCB and calpeptin, respectively, to protect neurons from beta-BuTx-induced cell death. These observations indicate that induction of cell death by beta-BuTx comprises several successive phases: (i) binding to neuronal potassium channels is the initial event, followed by (ii) internalization and (iii) induction of apoptotic cell death via a caspase-independent pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Annexin A5 / analysis
  • Apoptosis / drug effects*
  • Bungarotoxins / pharmacokinetics*
  • Calpain / antagonists & inhibitors
  • Calpain / metabolism
  • Caspase Inhibitors
  • Caspases / metabolism
  • Cells, Cultured
  • Cysteine Proteinase Inhibitors / pharmacology
  • Dipeptides / pharmacology
  • Dose-Response Relationship, Drug
  • Elapid Venoms / pharmacology
  • Female
  • Hippocampus / cytology
  • In Situ Nick-End Labeling
  • Neurons / chemistry
  • Neurons / cytology*
  • Neurons / metabolism
  • Neurotoxins / pharmacology
  • Phospholipases A / metabolism
  • Pregnancy
  • Rats
  • Rats, Wistar
  • Receptors, Cell Surface / metabolism*
  • Signal Transduction / physiology

Substances

  • Annexin A5
  • Bungarotoxins
  • Caspase Inhibitors
  • Cysteine Proteinase Inhibitors
  • Dipeptides
  • Elapid Venoms
  • Neurotoxins
  • Receptors, Cell Surface
  • dendrotoxin I
  • calpeptin
  • Phospholipases A
  • Calpain
  • Caspases