On the origin of ultraslow spontaneous Na+ fluctuations in neurons of the neonatal forebrain

Abstract

Spontaneous neuronal and astrocytic activity in the neonate forebrain is believed to drive the maturation of individual cells and their integration into complex brain-region-specific networks. The previously reported forms include bursts of electrical activity and oscillations in intracellular Ca²⁺ concentration. Here, we use ratiometric Na⁺ imaging to demonstrate spontaneous fluctuations in the intracellular Na⁺ concentration of CA1 pyramidal neurons and astrocytes in tissue slices obtained from the hippocampus of mice at postnatal days 2-4 (P2-4). These occur at very low frequency (∼2/h), can last minutes with amplitudes up to several millimolar, and mostly disappear after the first postnatal week. To further investigate their mechanisms, we model a network consisting of pyramidal neurons and interneurons. Experimentally observed Na⁺ fluctuations are mimicked when GABAergic inhibition in the simulated network is made depolarizing. Both our experiments and computational model show that blocking voltage-gated Na⁺ channels or GABAergic signaling significantly diminish the neuronal Na⁺ fluctuations. On the other hand, blocking a variety of other ion channels, receptors, or transporters including glutamatergic pathways does not have significant effects. Our model also shows that the amplitude and duration of Na⁺ fluctuations decrease as we increase the strength of glial K⁺ uptake. Furthermore, neurons with smaller somatic volumes exhibit fluctuations with higher frequency and amplitude. As opposed to this, larger extracellular to intracellular volume ratio observed in neonatal brain exerts a dampening effect. Finally, our model predicts that these periods of spontaneous Na⁺ influx leave neonatal neuronal networks more vulnerable to seizure-like states when compared with mature brain.NEW & NOTEWORTHY Spontaneous activity in the neonate forebrain plays a key role in cell maturation and brain development. We report spontaneous, ultraslow, asynchronous fluctuations in the intracellular Na⁺ concentration of neurons and astrocytes. We show that this activity is not correlated with the previously reported synchronous neuronal population bursting or Ca²⁺ oscillations, both of which occur at much faster timescales. Furthermore, extracellular K⁺ concentration remains nearly constant. The spontaneous Na⁺ fluctuations disappear after the first postnatal week.

Keywords: depolarizing GABA; hyperactivity; ion dynamics; neonatal brain; spontaneous Na+ fluctuations.