As a result of his investigation of the peripheral action of tetanus toxin, Harvey (1939) suggested that the condition of 'local tetanus' is caused by a disturbance of the normal processes of cholinergic transmission at the neuromuscular junction of skeletal muscle. The, effect of this toxin on smooth muscles innervated by cholinergic fibres does not appear to have been studied hitherto. We have begun such an investigation by observing the effects on the muscles of the iris that result from the injection of a small quantity of tetanus toxin into the anterior chamber of the eye. This has proved a satisfactory method of localizing the effect of the toxin, and has shown that this substance exerts a paralysing action on the cholinergic fibres to the sphincter pupillae.
MATERIALS AND METHODSDried preparations of ammonium sulphate-precipitated toxin were made from cultures of the CN655 strain of Cl. tetani by the method described by Ipsen (1941 a). The powdered toxin was kept in a vacuum desiccator, and fresh solutions of it in sterile 0*9 % NaCl were made shortly before each inoculation. No solution was more than i hr. old at the time that it was injected. The toxicity of the dried preparation was repeatedly tested by intravenous injection into mice and showed little deterioration over the period of the experiments. The LD50 of the dried toxin, calculated from Ipsen's (1941 b) Table 83, was 0 075 pg. The powder contained a small amount of (NH4)280 as an impurity.Before inoculation, the rabbits were lightly anaesthetized with 'veterinary nembutal' (0.75-1.5 c.c. intravenously), and, to ensure local insensibility, 2 drops of 5% cocaine-HCl were instilled into the conjunctival sac. When the cornea became anaesthetic, the eyeball was immobilized as much as possible by pressing on it from below with one finger placed on the medial half of the lower eyelid-the pressure being applied dorso-laterally towards the roof of the orbit. The carefully sharpened needle (size 26) of a tuberculin syringe was then introduced into the anterior chamber through the cornea just in front of the sclero-corneal junction at the inner or outer canthus (see Fig. 2, 2), and 0-05 c.c. of the solution was injected. The amount oftoxin varied from 0X25 to 200,ug., but in most of the experiments a standard dose of 25,ug. was adopted.In control experiments, the same volume of one of the following solutions was injected into the opposite eye: (1) toxin solution of the same concentration, in which the toxin had been neutralized by the previous addition of an amount of antitoxin (Wellcome tetanus antitoxin globulins: