R. Cahan et al., BIOCHEMICAL AND MORPHOLOGICAL-CHANGES IN RAT MUSCLE CULTURES CAUSED BY 28,000-MOL-WT TOXIN OF BACILLUS-THURINGIENSIS-ISRAELENSIS, Toxicon, 32(9), 1994, pp. 1125-1136
The 28,000 mol. wt protein of Bacillus thuringiensis israelensis showe
d a high degree of toxicity to rat muscle in culture. Application of 1
mu g/ml to the culture medium completely inhibited cell fusion. Rever
sibility of this effect was demonstrated by replacement of the culture
medium with fresh medium, and the consequence was that cell fusion wa
s resumed. When differentiated myotubes were treated with 1 mu g/ml of
the toxin, the spontaneous contractile activity was abolished within
20 min. Cytotoxic effects were observed 1 hr after treatment was initi
ated, as manifested by creatine kinase (CK) release to the medium. Two
hours after toxin was applied to the muscle culture, the myotubes wer
e deteriorated whereas the mononucleated cells were not affected. Six
or 7-day-old cultures which were treated by 1 mu g/ml of 28,00 mol. wt
toxin revealed a change in the levels of Na+ and K+ within the fibres
as analysed by X-ray microanalysis (XRMA). Preincubation of the toxin
for 20 min with phospholipids before application to the cells reduced
the cytotoxic effect. Phosphatidylinositol and phosphatidylserine wer
e the most efficient inhibitors, whereas phosphatidylcholine, sphingom
yelin and phosphatidylethanolamine were less effective in protecting c
ultures from the cytotoxic effects of the 28,000 mol. wt protein.