Morphine regulates DNA synthesis in rat cerebellar neuroblasts in vitro

The effects of morphine on DNA synthesis by external granular layer (EGL) neuroblasts was examined in whole-mount organotypic cultures isolated from 10-day-old rat cerebella using bromodeoxyuridine (BrdU). After 24 h in vitro, explants were treated for 24 h with 10 nM, 1 or 100 microM morphine, morphine plus 30 nM, 3 or 300 microM of the opiate antagonist naloxone, respectively, or those concentrations of naloxone alone. BrdU was added during the last 4 h of drug treatment. EGL neuroblasts were unambiguously identified by size and morphology, location and by protein kinase C II immunocytochemistry. The proportion of EGL neuroblasts incorporating BrdU was significantly reduced in the presence of 1 microM morphine, while 100 microM morphine had little additional effect. The concentration of morphine predicted to cause a half-maximal reduction in BrdU labeling index was 22.5 nM. Morphine's ability to reduce BrdU incorporation by EGL neuroblasts was concentration dependent and was prevented by concomitant treatment with naloxone, implicating the involvement of opioid receptors. The results suggest that morphine can directly regulate the growth of the developing cerebellum by inhibiting neuroblast proliferation within the EGL.