Reciprocal influences of nigral cells and striatal patch neurons in dissociated co-cultures.

ABSTRACT

Our previous work has shown that the functional efficacy of nigral tissue transplants into dopamine (DA)-depleted rats is increased when embryonic striatal tissue is included (Costantini et al. Exp Neurol 127219-231, 1994). To examine further the influence of striatal patch neurons in this regard, we employed co-cultures of dissociated nigral and striatal cells taken from embryos at different ages. Striatal patch neurons were labeled by in vivo bromodeoxyuridine (BrdU) on embryonic day (E)13 and E14. The percentage of striatal cells that were BrdU labeled was greater in E14 striatal cultures (51.0%) compared with E16 (33.9%) and E20 (3.5%) striatal cultures at 1 day in vitro (DIV). The proportion of surviving BrdU-labeled cells in striatal cultures decreased over time. The inclusion of E14 nigral cells attenuated this decline. Similarly, the number of dopaminergic [tyrosine hydroxylase (TH)-immunoreactive] neurons in pure nigral cultures decreased with time in vitro (8.2% at 1 DIV to 3.5% at 12-15 DIV). The inclusion of E14 striatal tissue increased the number of TH-immunoreactive neurons at all time points, whereas E16 and E20 striatal tissue was somewhat less effective. Thus, the survival of nigral DA neurons and striatal patch neurons in culture appears to be enhanced in the presence of the other. These reciprocal influences on neuronal survival may be relevant to the in vivo development of the nigrostriatal system as well as the enhanced function of cells in co-transplants.