Mechanism of Action of Bacillus thuringiensis Insecticidal Delta-endotoxin on Insect Cells in Vitro

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Cultured insect cells, TN-368 from the cabbage looper, swelled and burst upon treatment with the enzyme-digested delta-endotoxin of Bacillus thuringiensis var. aizawai. The cytotoxic sweelling was depended upon the amount of the delta-endotoxin added and the concentration of NaCl or KCl in the isotonic solution. The concentration of Na+ in the swollen cells approximately doubled in isotonic NaCl, while that of K+ decreased to 10% of the original cellular concentration. The cell swelling was inhibited by tetrodotoxin and also by ouabain in only KCl isotonic solution. On the other hand, 4-aminopyridine stimulated the swelling in the isotonic KCl solution, These results indicate that the delta-endotoxin induces the stimulation of Na+ influx and K+ efflux in the isotonic NaCl solution, and also stimulates the Na+, K+-ATPase in the isotonic KCl solution. The cytotoxic swelling was also blocked by cAMP, AMP, ATP, GTP, and NAD, but not by adenosine and GMP. These results suggest the participation of nucleotide derivatives in the action of delta-endotoxin.