Survival promotion of rat cerebellar granule neurons by co-culture with pontine explant

Cultured cerebellar granule neuron (CGN) of the rat is the most frequently used model system for analysis of activity-dependent neuronal survival. CGNs do not survive longer than 2 weeks in a standard culture medium unless KCl (or other excitants such as glutamate) is added. It is assumed that KCl represents synaptic activity, but no tests have been made on whether the survival of CGNs really depends on the synaptic input. Here we co-cultured CGNs with an explant of the pons including the basilar pontine nucleus (BPN), which is one of the input sources of CGNs in vivo, to confirm if synaptic input is really a determinant for the survival of these cells. In this co-culture system, the viability of CGNs was significantly increased without the addition of KCl. The survival promotion was confined to the population of CGNs having contact with neurites of BPN and was cancelled by an application of tetrodotoxin or antagonists of glutamate receptors, indicating that the survival depended on synaptic activity. Explants of other glutamatergic tissues including the hippocampus failed to promote the survival, although neurites grew out from these explants as vigorously as from the BPN explants. Calcium and FM1-43 imaging examinations revealed that the CGNs had formed functional synapses with the BPN explant but not with the hippocampal explant. These results, confirming the assumption that synaptic activity determines neuronal survival, provide evidence for presynaptic contribution to the survival. [Copyright &y& Elsevier]