title: The rat Digit Abduction Score (DAS) assay: a physiological model for assessing botulinum neurotoxin-induced skeletal muscle paralysis.
abstract: Botulinum neurotoxins (BoNT) are approved for a number of therapeutic indications, including blepharospasm, cervical dystonia and hyperhidrosis, and have also shown efficacy in a variety of pain disorders. The potency of any given BoNT preparation can be routinely assessed by using the Digit Abduction Score (DAS) assay, which measures the local muscle weakening efficacy of BoNT following injection into mouse hindlimb muscle. While most studies have employed mice to assess BoNT efficacy in the DAS, few have utilized rats. In this study, we applied the DAS assay to a rat model and compared the potency of IM-BOTOX(®) (onabotulinumtoxinA) injections between two separate hind limb muscles, gastrocnemius and tibialis anterior (TA). The results demonstrated that the DAS assay can be performed on rats with similar criteria and parameters as for mice. Moreover in the rat, BoNT can be injected into either the gastrocnemius or TA muscle to elicit similar DAS scoring responses. Interestingly, onabotulinumtoxinA potency in the rat DAS was ∼3-fold higher following TA injections than gastrocnemius injections. Additionally, our data showed that the durational kinetics of onabotulinumtoxinA in the rat DAS are approximately twice as long as in the mouse DAS. These results position the rat DAS as a more flexible model for examining the mechanisms of BoNT diffusion and muscle paralysis, while mouse DAS can be used for physiological screening of BoNT because of the potential for higher throughput. Overall, these data confirm the utility of the DAS assay for characterizing the physiological potency of BoNT and related compounds.
authors.affiliation: Allergan, Inc., LLC, 2525 Dupont Drive, Irvine, CA 92612-1599, USA. broide_ron@allergan.com
authors.forename: Ron S
authors.initials: RS
authors.lastname: Broide
authors.forename: John
authors.initials: J
authors.lastname: Rubino
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Nicholson
authors.forename: Maria C
authors.initials: MC
authors.lastname: Ardila
authors.forename: Meenakshi S
authors.initials: MS
authors.lastname: Brown
authors.forename: K Roger
authors.initials: KR
authors.lastname: Aoki
authors.forename: Joseph
authors.initials: J
authors.lastname: Francis
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D019274:Botulinum Toxins, Type A / Q000493:pharmacokinetics / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005260:Female; D006614:Hindlimb / Q000187:drug effects; D007273:Injections, Intramuscular; D008297:Male; D051379:Mice; D023421:Models, Animal; D018482:Muscle, Skeletal / Q000187:drug effects*; D019148:Neuromuscular Blockade; D009498:Neurotoxins / Q000493:pharmacokinetics / Q000633:toxicity; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D009498:Neurotoxins; D019274:Botulinum Toxins, Type A
keywords: BoNT; Botulinum neurotoxin; DAS; Digit Abduction Score; Gastrocnemius muscle; LABORAS; Laboratory Animal Behavior Observation, Registration and Analysis System; OnabotulinumtoxinA; TA; Tibialis anterior muscle; botulinum neurotoxin; tibialis anterior