Primary cell culture for evaluation of botulinum neurotoxin antagonists.
The actions of botulinum neurotoxin (BoNT) were studied on evoked release of the neurotransmitter glycine in primary mouse spinal cord cells. 3[H]-glycine was taken up by cells in physiological solution and released by depolarization with 56 mM K+ in the presence of 2 mM Ca2+. Release of 3[H]-glycine was found to be inhibited by BoNT serotypes A, B and E with similar potency ratios to those observed in the acutely isolated mouse diaphragm muscle. When spinal cord cultures were exposed to BoNT/A for 24 h, inhibition of 3[H]-glycine release was detected at toxin concentrations as low as 10(-14) M, and complete inhibition was observed at concentration >or=10(-12) M. Preincubation of BoNT/A with polyclonal equine antiserum led to antagonism of toxin-induced inhibition of 3[H]-glycine release in spinal cord cells and to protection of mice from the lethal effects of BoNT/A. It is concluded that spinal cord neurons are a useful model for studying botulinum intoxication and for evaluating BoNT antagonists.
Sheridan
Robert E
RE
Neurotoxicology Branch, Pharmacology Division, US Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, MD 21010-5400, USA.
Smith
Theresa J
TJ
Adler
Michael
M
D000818:Animals; D001905:Botulinum Toxins / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D002478:Cells, Cultured; D005333:Fetus; D005998:Glycine / Q000378:metabolism; D007106:Immune Sera / Q000494:pharmacology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology* / Q000196:embryology
D007106:Immune Sera; D001905:Botulinum Toxins; D005998:Glycine