Early postnatal maturation of GABA(A)-mediated inhibition in the brainstem respiratory rhythm-generating network of the mouse

Abstract

It is well established that GABA(A)-mediated postsynaptic potentials are excitatory in many brain regions during embryonic and early postnatal life. The pre-Botzinger complex (PBC) in the brainstem is an essential component of the respiratory rhythm-generating network, where GABA(A)-mediated inhibition plays a critical role in generating a stable respiratory rhythm in adult animals. In the present study, using the perforated patch technique, we investigated the maturation of GABA(A) receptor-mediated effects on rhythmically active PBC neurons and on the motor output in slice preparations from P0-15 neonatal mice. The reversal potential of GABA(A) receptor-mediated current (EGABA-A) switched from depolarizing to hyperpolarizing within the first postnatal week. EGABA-A was -13.7 +/- 9.8 mV at P0, then it changed to -44.8 +/- 7.0 mV at P2 and -71.5 +/- 6.8 mV at P4. Perfusion of bicarbonate-free saline has no detectable influence on EGABA-A, indicating that a lack of Cl- extrusion during P0-3 is mainly responsible for early GABA(A)-ergic excitation. At the network level, blockade of GABA(A) receptors with bicuculline did not significantly change the frequency of rhythmic bursts recorded from hypoglossal nerve roots before P3, whereas it increased the coefficient of variation. After P3, bicuculline increased burst frequency with little effect on the coefficient of variation. Thus, chloride-mediated inhibition, which appears in PBC neurons after P3, coincides with the appearance of GABA(A)-mediated modulation of the respiratory rhythm. GABA(A) receptor-activated inhibition may therefore be necessary for frequency modulation in the respiratory network beginning on the fourth postnatal day in the mouse brainstem.