1. Cellular responses to GABA(A) receptor activation were studied in develo
ping cerebellar Purkinje neurones (PNs) in brain slices obtained from 2- to
22-day-old rats. Two-photon fluorescence imaging of fura-2-loaded cells an
d perforated-patch recordings were used to monitor intracellular Ca2+ trans
ients and to estimate the reversal potential of GABA-induced currents, resp
ectively.
2. During the 1st postnatal week, focal application of GABA or the GABA(A)
receptor agonist muscimol evoked transient increases in [Ca2+](i) in immatu
re PNs. These Ca2+ transients were reversibly abolished by the GABAA recept
or antagonist bicuculline and by Ni2+, a blocker of voltage-activated Ca2channels.
3. Perforated-patch recordings were used to measure the reversal potential
of GABA-evoked currents (E-GABA) at different stages of development. It was
found that E-GABA was about -44 mV at postnatal day 3 (P3), it shifted to
gradually more negative values during the 1st week and finally equilibrated
at -87 mV at around the end of the 2nd postnatal week. This transition was
well described by a sigmoidal function. The largest change in EGABA was -7
mV day(-1), which occurred at around P6.
4. The transition in GABA-mediated signalling occurs during a period in whi
ch striking changes in PN morphology and synaptic connectivity are known to
take place. Since such changes were shown to be Ca2+ dependent, we propose
that GABA-evoked Ca2+ signalling is one of the critical determinants for t
he normal development of cerebellar PNs.