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Fig. 3. Effect of blockade of NO production on cell proliferation. (A) Cell proliferation was measured on cerebellar homogenates from rat pups treated with L-NAME (60 mg/kg/day) or with ODQ (5 mg/kg/day) for various neonatal intervals (P1-P2, P1-P3, P3-P5, P5-P7), while control pups received vehicle. Two hours prior to sacrifice, pups were s.c. injected with [³H]thymidine (5 µCi/g body weight). The % of [³H]thymidine incorporation into DNA was expressed as dpm/µg protein. Values are the mean ± s.e.m. of four different experiments ^*P<0.05, ^**P<0.01 compared with the corresponding controls of the same age (dotted line) (Bonferroni's test after ANOVA). (B) To assess apoptosis in the developing cerebellum, cerebellar homogenates from pups of various postnatal ages (3, 5, 7 and 11 days) were monitored, using a sandwich ELISA method, for histone-associated DNA fragments in the cytoplasm. L-NAME-treated pups received the usual drug dosage at the following intervals: P1-P3, P3-P5, P5-P7, P9-P11. Values are the mean ± s.e.m. of three independent experiments (Bonferroni's test after ANOVA). (C,D) BrdU-labelled cells in the cerebellum of 3-day-old pups after BrdU administration at P3, in a control rat (C) and a rat treated with L-NAME from P1 to P3 (D). Bar, 100 µm. (E) Density of BrdU-labelled cells (number of cells/mm²) in the external granular layer (EGL) and internal granular layer (IGL) of the cerebellum of control and L-NAME P1-P3 treated 3-day-old rat pups. Values are the mean ± s.e.m. of three experiments. ^*P<0.01, compared with the control (Student's t-test).

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