Intrinsic brainstem circuits comprised of Chx10-expressing neurons contribute to reticulospinal output in mice

Glutamatergic reticulospinal neurons in the gigantocellular reticular nucleus (GRN) of the medullary reticular formation can function as command neurons, transmitting motor commands to spinal cord circuits to instruct movement. Recent advances in our understanding of this neuron-dense region have been facilitated by the discovery of expression of the transcriptional regulator, Chx10, in excitatory reticulospinal neurons. Here, we address the capacity of local circuitry in the GRN to contribute to reticulospinal output. We define two sub-populations of Chx10-expressing neurons in this region, based on distinct electrophysiological properties and somata size (small and large), and show that these populations correspond to local interneurons and reticulospinal neurons, respectively. Using focal release of caged glutamate combined with patch clamp recordings, we demonstrated that Chx10 neurons form microcircuits in which the Chx10 local interneurons project to and facilitate the firing of Chx10 reticulospinal neurons. We discuss the implications of these microcircuits in terms of movement selection.

Distributed and diverse hindbrain neuronal activity contributes to sensory processing and motor control in the Xenopus laevis tadpole

Locomotion is a key feature of healthy animals, which depends on their ability to move -or not to move- for their survival. The hatchling Xenopus laevis tadpole responds to trunk skin stimulation by swimming away, and its developing nervous system is simple enough to make it an ideal model organism to study the control of locomotion. This vertebrate embryo relies on excitatory cells in the skin to detect the sensory stimulus, which is quickly sent to the brain via ascending sensory pathway neurons. When the stimulation is strong enough, descending reticulospinal neurons are activated in the hindbrain and the spinal cord, after a long and variable delay. The activation of reticulospinal neurons indicates the initiation of swimming and sustains the rhythmic firing of CPG (central pattern generator) neurons, among which are motor neurons. The tadpole is then able to rhythmically contract trunk muscles, allowing the undulatory movement of swimming. However, how the tadpole developing brain exerts descending control over reticulospinal neurons, and thus over the spinal CPG centers, is not fully understood yet. In this work, we recorded extracellular activity in the hindbrain of the tadpole to identify firing units that are involved in the long and variable delay to swim initiation following trunk skin stimulation. We isolated firing units that mediate distinct motor output. We subsequently grouped them in populations based on their firing patterns in response to skin stimulation and motor output. We propose a novel neural circuitry for sensory processing and descending motor control exerted by the hindbrain of the hatchling tadpole, which could account for the long and variable delay to reticulospinal neuron activation, and thus swim initiation.

Local brainstem circuits contribute to reticulospinal output in the mouse

ABSTRACTGlutamatergic reticulospinal neurons in the gigantocellular reticular nucleus (GRN) of the medullary reticular formation can function as command neurons, transmitting motor commands to spinal cord circuits. Recent advances in our understanding of this neuron-dense region have been facilitated by the discovery of expression of the transcriptional regulator, Chx10, in excitatory reticulospinal neurons. Here, we address the capacity of local circuitry in the GRN to contribute to reticulospinal output. We define two sub-populations of Chx10-expressing neurons in this region, based on distinct electrophysiological properties and somata size (small and large), and show that these correspond to local interneurons and reticulospinal neurons, respectively. Using focal release of caged-glutamate combined with patch clamp recordings, we demonstrated that Chx10 neurons form microcircuits in which the Chx10 interneurons project to and facilitate the firing of Chx10 reticulospinal neurons. We discuss the implications of these microcircuits in terms of movement selection.SIGNIFICANCE STATEMENTReticulospinal neurons in the medullary reticular formation play a key role in movement. The transcriptional regulator Chx10 defines a population of glutamatergic neurons in this region, a proportion of which have been shown to be involved in stopping, steering, and modulating locomotion. While it has been shown that these neurons integrate descending inputs, we asked whether local processing also ultimately contributes to reticulospinal outputs. Here, we define Chx10-expressing medullary reticular formation interneurons and reticulospinal neurons, and demonstrate how the former modulate the output of the latter. The results shed light on the internal organization and microcircuit formation of reticular formation neurons.

The Lamprey’s Dilemma

This chapter focuses on the neural network, demonstrating how the principles described in the previous chapter are implemented in vertebrates, taking as a blueprint the oldest one: the lamprey. The reticulospinal neurons belong to the reticular formation located in the brainstem of the lamprey. These reticulospinal neurons act as the effector system. Apart from the peripheral input that comes back from the spinal cord, the reticular formation receives, among other things, input from the diencephalon and specifically the thalamus. This structure allows interfacing between sensory stimuli (visual, auditory, and olfactory) and the motor system. The other very important targets of the thalamus in the lamprey are the basal ganglia. The chapter then goes on to explain the diencephalic and telencephalic loops.

Modeling of lamprey reticulospinal neurons: multiple distinct parameter sets yield realistic simulations

A computer model of lamprey reticulospinal neurons with a default parameter set produced simulations of action potentials and repetitive firing that scored favorably compared with the properties of these neurons. A dual-annealing search algorithm explored ~50 million parameter sets and identified 4,302 distinct viable parameter sets within parameter space that yielded higher/much higher scores than the default parameter set. In addition, 5- and 2-conductance grid searches identified significant correlations between maximum conductances for pairs of ion channels.

The serotonin reuptake blocker citalopram destabilizes fictive locomotor activity in salamander axial circuits through 5-HT1A receptors

Serotoninergic (5-HT) neurons are powerful modulators of spinal locomotor circuits. Most studies about 5-HT modulation focused on the effect of exogenous 5-HT and these studies provided key information about the cellular mechanisms involved. Less is known about the effects of increased release of endogenous 5-HT with selective serotonin reuptake inhibitors. Such molecules were shown to destabilize the locomotor output of spinal limb networks through 5-HT1A receptors. However, in tetrapods little is known about the effects of increased 5-HT release on the locomotor output of axial networks, which are coordinated with limb circuits during locomotion from basal vertebrates to mammals. Here, we examined the effect of citalopram on fictive locomotion generated in axial segments of isolated spinal cords in salamanders, a tetrapod where raphe 5-HT reticulospinal neurons and intraspinal 5-HT neurons are present as in other vertebrates. Using electrophysiological recordings of ventral roots, we show that fictive locomotion generated by bath-applied glutamatergic agonists is destabilized by citalopram. Citalopram-induced destabilization was prevented by a 5-HT1A receptor antagonist, whereas a 5-HT1A receptor agonist destabilized fictive locomotion. Using immunofluorescence experiments, we found 5-HT-positive fibers and varicosities in proximity with motoneurons and glutamatergic interneurons that are likely involved in rhythmogenesis. Our results show that increasing 5-HT release has a deleterious effect on axial locomotor activity through 5-HT1A receptors. This is consistent with studies in limb networks of turtle and mouse, suggesting that this part of the complex 5-HT modulation of spinal locomotor circuits is common to limb and axial networks in limbed vertebrates.

Brainstem Neurons that Command Left/Right Locomotor Asymmetries

Descending command neurons instruct spinal networks to execute basic locomotor functions, such as which gait and what speed. The command functions for gait and speed are symmetric, implying that a separate unknown system directs asymmetric movements—the ability to move left or right. Here we report the discovery that Chx10-lineage reticulospinal neurons act to control the direction of locomotor movements in mammals. Chx10 neurons exhibit ipsilateral projection, and can decrease spinal limb-based locomotor activity ipsilaterally. This circuit mechanism acts as the basis for left or right locomotor movements in freely moving animals: selective unilateral activation of Chx10 neurons causes ipsilateral movements whereas inhibition causes contralateral movements. Spontaneous forward locomotion is thus transformed into an ipsilateral movement by braking locomotion on the ipsilateral side. We identify sensorimotor brain regions that project onto Chx10 reticulospinal neurons, and demonstrate that their unilateral activation can impart left/right directional commands. Together these data identify the descending motor system which commands left/right locomotor asymmetries in mammals.

Spinal Circuit for Escape in Goldfish and Zebrafish

Escape or startle responses are vital to organisms. In fishes, escape behavior is a rapid bend of the body and tail away from a potential threat that occurs within milliseconds after a stimulus. When properly executed, it is a fast, powerful body bend to only one side that takes precedence over any other movements. The behavior is initiated by the firing of one of a bilateral pair of hindbrain reticulospinal neurons (RSNs) called Mauthner cells (M-cells). The output of each cell occurs via an axon that crosses in the brain and extends the length of the spinal cord on the opposite side of the body. The circuit of the M-cell in spinal cord is based upon studies of goldfish and zebrafish. This circuit, repeated along the spinal cord, has several features that are well matched to the behavioral demands of escape movements.