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Related Experiment Videos

Ionic permeabilities induced by Bacillus thuringiensis in Sf9 cells

V Vachon1, M J Paradis, M Marsolais

  • 1Groupe de recherche en transport membranaire, Université de Montréal, Quebec, Canada.

The Journal of Membrane Biology
|November 1, 1995
PubMed
Summary
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Bacillus thuringiensis insecticidal toxins, like CryIC, rapidly increase cell membrane permeability in fall armyworm cells. This leads to the efflux of ions such as potassium (K+) and sodium (Na+), altering intracellular pH.

Area of Science:

  • Molecular Biology
  • Insect Toxicology
  • Cell Physiology

Background:

  • Bacillus thuringiensis (Bt) produces insecticidal toxins.
  • Sf9 cells from Spodoptera frugiperda (fall armyworm) are a model system for lepidopteran research.
  • Intracellular pH (pHi) regulation is crucial for cell function.

Purpose of the Study:

  • To investigate the effect of Bt insecticidal toxins on monovalent cation content and pHi in Sf9 cells.
  • To determine the specificity of Bt toxins on ion permeability and pHi regulation.

Main Methods:

  • Utilized fluorescent indicators PBFI and BCECF to monitor K+, Na+, and pHi in Sf9 cells.
  • Exposed Sf9 cells to specific Bt toxins (CryIC, CryIIIA, CryIA variants) under varying medium conditions.
  • Analyzed changes in ion flux and pHi using fluorescence ratio measurements.

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Main Results:

  • CryIC toxin induced rapid efflux of K+ and Na+ from Sf9 cells.
  • CryIC toxin caused Sf9 cells' pHi to rapidly equilibrate with the external solution, dependent on time and concentration.
  • Other Bt toxins (CryIIIA, CryIA variants) showed no significant effect on pHi in Sf9 cells.

Conclusions:

  • Bt endotoxins, specifically CryIC, act by increasing the permeability of the cytoplasmic membrane to H+, K+, and Na+ in susceptible insect cells.
  • This ion permeability increase disrupts normal cellular ion balance and pHi regulation, contributing to insecticidal activity.
  • Bt toxin activity is specific, with CryIC targeting lepidopteran cells effectively.