The relationship between synaptogenesis and expression of voltage-dependent currents in cerebellar granule cells in situ.

In this work we consider the ontogenetic changes of membrane currents and their relationship with synaptogenesis in cerebellar granule cells. Recordings were performed in whole-cell patch-clamp configuration from cerebellar slices obtained from 4 to 31-day-old rats. Granule cells in the external granular layer, and non-connected granule cells in the internal granular layer expressed outward currents, and inconstantly also small Ca2+ currents, but no fast Na+ currents. Most connected granule cells expressed Ca2+ and Na+ currents. These data indicate that Ca2+ and Na+ current development occurs after synapse formation, while outward (K+) currents begin their development before. Mixed NMDA/non-NMDA synaptic currents were observed at all stages, while synaptic currents with a prominent NMDA component were observed exclusively at immature stages. At P4, ie 1-2 days after the arrival of the first granule cells in the internal granular layer, some granule cells already expressed mature synaptic and voltage-dependent currents, suggesting that establishment of mossy fibre synapses and development of membrane properties takes just 1-2 days to complete. Starting at P4, the probability of activating mossy fibre currents, and sizeable Ca2+ and Na+ currents increased at a similar rate, attaining a plateau level around P20. Average amplitude of Na+ and outward currents decreased until P10 and then increased attaining plateau soon beyond P20. Average amplitude of Ca2+ currents increased monotonically. The time courses of probability and average current amplitude curves are likely explained by changes in the rate of accumulation of migrating granule cells in the internal granular layer, and by changes in granule cell membrane surface extension. These data suggest a relevant role for the process of synapse formation in inducing the expression of new channels in the developing granule cells, which may involve Ca2+ influx through the NMDA channel.