Neuronal hyperactivity sustains the basal dendrites of immature dentate granule cells: time-lapse confocal analysis...

Dendritic morphogenesis is an essential process for the establishment of proper neural circuitry. In the epileptic hippocampus, mature dentate granule cells (GCs) possess basal dendrites (BDs), which is abnormal and is assumed to contribute to seizure progression. However, there is a lack of direct time-lapse evidence showing that neuronal hyperactivity regulates the dendritic development of GCs. In the present study, we carried out time-lapse confocal analysis of the dendritic morphogenesis of GCs in hippocampal slice cultures that were prepared from postnatal 6-day-old (P6) rats. By electroporating membrane-targeted green fluorescent protein at 5 days in vitro (DIV), we found that most of the Prox1-positive and calbindin-negative immature GCs possessed several BDs and filopodia-rich apical dendrites at 7 DIV. BDs were gradually eliminated from 7 to 9 DIV, and they completely vanished at 14 DIV in all the GCs examined. However, most BDs failed to retract from 7 to 9 DIV, when the GABA(A) receptor antagonist picrotoxin was chronically applied to induce epileptic conditions in the cultures. These effects were blocked by coapplying tetrodotoxin, a sodium channel blocker, thus convincing us that neuronal hyperactivity contributes to the maintenance of BDs. Further, in the picrotoxin-treated cultures, most of the GCs persistently exhibited several BDs even after 14 DIV. In contrast, neither the progressive pruning of the filopodia nor the branch dynamics of the apical dendrites during the culture periods was affected by picrotoxin. These results, for the first time, provide us with direct evidence that neuronal hyperactivity contributes to the stability of pre-existing BDs.