The organization ofponto-neocerebellar projections in the cat 4 has been recently re-examined with tracer techniquesT,11,17. Within the framework of the topographical organization, considerable overlap was found between sets of neurons which project to different subdivisions of the cerebellar hemisphere and neocerebellar vermis (lobulus Vlla, b4,11,17. In addition, projections from each half of pontine nuclear complex (PN) to lateral parts of crus I-II were found to be relatively more bilateral than hitherto described 17. In the present study, combinations of two fluorescent retrograde tracers1, 2 13,14,19,z0 were used to determine whether this bilaterality 17 could be accounted for by divergent axon collaterals given off by individual pontine neurons. In preliminary experiments the best results were obtained with 1 .5~ (w/v) bisbenzimide (Bb; Hoechst 33258) in distilled water, 10~ (w/v) Evans blue (EB; Merck 3169) in 1 700 poly-L-ornithine in distilled water or 5 ~ (w/v) 'Granular blue' (GB; compound 184/134)* in distilled water with survival times of 8-10 11 (Bb), 3 days (EB) or 9-10 days (GB). Therefore, in one group of 7 cats, 3 were injected with 10 ~ EB (0.8-1.6 /A) combined with 5 ~ GB (1.2-2.6 #1) and the other 3 cats with EB combined with 1.5 ~ Bb (0.6-1.2 #1). The tracers were injected into the lateral crus II, one on each side. In one experiment the cerebellum was transected at the midline 47 days before the EB and GB injections. In a second group of 3 cats, GB was injected into the vermal lobule VIIa,b and EB into one lateral crus II. At the various survival times, the animals were deeply anesthetized with Nembutal and perfused with 10~ formalin. The brain stem and cerebellum were impregnated with 30 ~ sucrose in 0. 2 M sodium cacodylate buffer, at 4 °C and then frozen serial sections, 25 #m frontal for the brain stem and 50 #m horizontal for the cerebellum, were cut. The sections were mounted on slides without coverslips,
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