in-vivo-visualization-spontaneous-activity-neonatal-mouse-sensory

In Vivo Visualization of Spontaneous Activity in Neonatal Mouse Sensory Cortex at a Single-Neuron Resolution

The mammalian brain undergoes dynamic developmental changes at both the cellular and circuit levels throughout prenatal and postnatal periods. Following ...

Kumamoto University

Cedars-Sinai Medical Center

The cerebral cortex has complex yet perfectly wired neuronal circuits that are important for high-level brain functions such as perception and cognition. The rodent's somatosensory system is widely used for understanding the mechanisms of circuit formation during early developmental periods. In this review, we summarize the developmental processes of circuit formation in layer 4 of the somatosensory cortex, and we describe the molecules involved in layer 4 circuit formation and neuronal activity-dependent mechanisms of circuit formation. We also introduce the dynamic mechanisms of circuit formation in layer 4 revealed by intravital two-photon imaging technologies, which include time-lapse imaging of neuronal morphology and calcium imaging of neuronal activity in newborn mice.

Elucidating mechanisms of neuronal circuit formation in layer 4 of the somatosensory cortex via intravital imaging.

Correlated spontaneous activity plays critical role in the organization of neocortical circuits during development. However, cortical mechanisms regulating activity correlation are still elusive. In this study, using two-photon calcium imaging of the barrel cortex layer 4 (L4) in living neonatal mice, we found that NMDA receptors (NMDARs) in L4 neurons are important for enhancement of spontaneous activity correlation. Disruption of GluN1 (), an obligatory NMDAR subunit, in a sparse population of L4 neurons reduced activity correlation between GluN1 knock-out (GluN1KO) neuron pairs within a barrel. This reduction in activity correlation was even detected in L4 neuron pairs in neighboring barrels and most evident when either or both of neurons are located on the barrel edge. Our results provide evidence for the involvement of L4 neuron NMDARs in spatial organization of the spontaneous firing activity of L4 neurons in the neonatal barrel cortex. Precise wiring of the thalamocortical circuits is necessary for proper sensory information processing, and thalamus-derived correlated spontaneous activity is important for thalamocortical circuit formation. The molecular mechanisms involved in the correlated activity transfer from the thalamus to the neocortex are largely unknown.  two-photon calcium imaging of the neonatal barrel cortex revealed that correlated spontaneous activity between layer four neurons is reduced by mosaic knock-out (KO) of the NMDA receptor (NMDAR) obligatory subunit GluN1. Our results suggest that the function of NMDARs in layer four neurons is necessary for the communication between presynaptic and postsynaptic partners during thalamocortical circuit formation.

NMDA Receptor Enhances Correlation of Spontaneous Activity in Neonatal Barrel Cortex.

The cerebral cortex comprises a complex and exquisite network of neuronal circuits that is formed during development. To explore the molecular mechanisms involved in cortical circuit formation, the tactile somatosensory pathway that connects the whiskers and cortex of rodents is a useful model. Here, we analyzed the roles of Ras GTPase-activating proteins (RasGAPs) in the circuit formation in the somatosensory cortex layer 4 (L4). We suppressed the function of RasGAPs in L4 neurons using Supernova RNAi, a plasmid vector-based sparse cell gene knockdown (KD) system. The results showed disrupted dendritic pattern formation of L4 spiny stellate neurons on the barrel edge by RasGAP KD. Furthermore, the number of presynaptic boutons on L4 neurons was reduced by RasGAP KD. These results demonstrate the essential roles of RasGAPs in circuit formation in the cerebral cortex and imply that developmental changes in dendrites and synapses in RasGAP KD neurons may be related to cognitive disabilities in RasGAP-deficient individuals, such as patients with neurofibromatosis type 1.

Madhura S. Rao

Laboratory of Multi-Dimensional Imaging,
International Research Center for Medical Sciences (IRCMS),
Graduate School of Medical Sciences,
Laboratory of Multi-Dimensional Imaging, International Research Center for Medical Sciences (IRCMS)

Kumamoto University

In Vivo Visualization of Spontaneous Activity in Neonatal Mouse Sensory Cortex at a Single-Neuron Resolution