Quantitative determination of biological activity of botulinum toxins utilizing compound muscle action potentials (CMAP), and comparison of neuromuscular transmission blockage and muscle flaccidity among toxins.
The biological activity of various types of botulinum toxin has been evaluated using the mouse intraperitoneal LD(50) test (ip LD(50)). This method requires a large number of mice to precisely determine toxin activity, and so has posed a problem with regard to animal welfare. We have used a direct measure of neuromuscular transmission, the compound muscle action potential (CMAP), to evaluate the effect of different types of botulinum neurotoxin (NTX), and we compared the effects of these toxins to evaluate muscle relaxation by employing the digit abduction scoring (DAS) assay. This method can be used to measure a broad range of toxin activities the day after administration. Types A, C, C/D, and E NTX reduced the CMAP amplitude one day after administration at below 1 ip LD(50), an effect that cannot be detected using the mouse ip LD(50) assay. The method is useful not only for measuring toxin activity, but also for evaluating the characteristics of different types of NTX. The rat CMAP test is straightforward, highly reproducible, and can directly determine the efficacy of toxin preparations through their inhibition of neuromuscular transmission. Thus, this method may be suitable for pharmacology studies and the quality control of toxin preparations.
The Chemo-Sero-Therapeutic Research Institute (KAKETSUKEN), 1-6-1 Okubo, Kumamoto-shi, Kumamoto 860-8568, Japan. torii-ya@kaketsuken.or.jp
Yasushi
Y
Torii
Yoshitaka
Y
Goto
Motohide
M
Takahashi
Setsuji
S
Ishida
Tetsuhiro
T
Harakawa
Takashi
T
Sakamoto
Ryuji
R
Kaji
Shunji
S
Kozaki
Akihiro
A
Ginnaga
D000200:Action Potentials / Q000187:drug effects*; D000818:Animals; D001905:Botulinum Toxins / Q000737:chemistry / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005260:Female; D007928:Lethal Dose 50; D051379:Mice; D008813:Mice, Inbred ICR; D009125:Muscle Relaxants, Central / Q000494:pharmacology; D018482:Muscle, Skeletal / Q000187:drug effects*; D009466:Neuromuscular Blocking Agents / Q000633:toxicity*; D009435:Synaptic Transmission / Q000187:drug effects*
D009125:Muscle Relaxants, Central; D009466:Neuromuscular Blocking Agents; D001905:Botulinum Toxins