Locomotion, including running, walking, and swimming, is a complex behavior enabling animals to interact with the environment. Vertebrate locomotion depends upon sets of interneurons in the spinal cord, known as the central pattern generator (CPG). The CPG performs multiple roles: pattern formation (left-right alternation and flexor-extensor alternation) and rhythm generation (the onset and frequency of locomotion). Many studies have begun to unravel the organization of the neuronal circuits underlying left-right and flexor-extensor alternation. However, despite pharmacologic, lesion, and optogenetic studies suggesting that the rhythm generating neurons are ispilaterally-projecting glutamatergic neurons, the precise cellular identification of rhythm generating neurons remains largely unknown.
Traditionally, CPG networks (both pattern formation and rhythm generation) are thought to reside upstream of motor neurons, which serve as the output of the spinal cord. Recently however, it has been discovered that direct stimulation of lumbar motor neurons using the intact ex vivo neonate mouse spinal cord preparation can activate CPG networks to produce locomotor-like behavior. Furthermore, depressing motor neuron discharge decreases locomotor frequency, whereas increasing motor neuron discharge accelerates locomotor frequency, suggesting that motor neurons provide ongoing feedback to the CPG. However, the circuit mechanisms through which motor neurons can influence activity in the CPG in mammals remain unknown.
Here, I used motor neurons as a means of accessing CPG interneurons by asking how motor neuron activation might induce locomotor-like activity. Through intracellular recording and morphological assays, I discovered that ventral spinocerebellar tract (VSCT) neurons are activated monosynaptically following motor neuron axon stimulation through chemical and electrical synapses. A subset of VSCT neurons were located close to or within the motor neuron nucleus. VSCT neurons were found to be excitatory, have descending spinal axon collaterals, and influence motor neuron output, suggesting that VSCT neurons are positioned advantageously to initiate and maintain locomotor-like rhythmogenesis. Intracellular recording from VSCT neurons revealed that they exhibit rhythmic activity during locomotor-like activity. VSCT neurons were found to contain the rhythmogenic pacemaker Ih current and to be connected to other VSCT neurons, at least through gap junctions. Optogenetic and chemogenetic manipulation of VSCT neuron activity provided evidence that VSCT neurons are both necessary and sufficient for the production of locomotor-like activity. Silencing VSCT neurons prevented the induction of such activity, whereas activation of VSCT neurons was capable of inducing locomotor-like activity. The production of locomotor-like activity by VSCT neuron photoactivation was dependent upon both electrical communication through gap junctions as well as the pacemaker Ih current.
The evidence presented in this thesis suggests that VSCT neurons are critical components for rhythm generation in the mammalian CPG and are key mediators of locomotor activity.
| Identifer | oai:union.ndltd.org:columbia.edu/oai:academiccommons.columbia.edu:10.7916/d8-yths-d668 |
| Date | January 2019 |
| Creators | Chalif, Joshua |
| Source Sets | Columbia University |
| Language | English |
| Detected Language | English |
| Type | Theses |
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