Thesis (Ph. D.)--University of Oregon, 2004. / Typescript. Includes vita and abstract. Includes bibliographical references (leaves 83-91). Also available for download via the World Wide Web; free to University of Oregon users.

Thesis (Ph.D.)--University of Oregon, 2005. / Typescript. Includes vita and abstract. Includes bibliographical references (leaves 96-103). Also available for download via the World Wide Web; free to University of Oregon users.

published_or_final_version / abstract / Medicine / Master / Master of Research in Medicine

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.

"April 2003" / Bibliography: leaves 124-144. / xvii, 144 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, School of Molecular and Biomedical Sciences, Discipline of Physiology, 2003

All motor behaviours are expressed via the activation of alpha motoneurones, the final common path of the central nervous system. The corticospinal tract conveys neural information from the motor cortex to motoneurones. This thesis focuses on the corticospinal control of human motoneurones during voluntary contraction and fatigue. First, output of motoneurones to corticospinal inputs is described for a wide range of contraction strengths. Results show that motoneurones become less responsive during strong contractions whereas motor cortical output cells are not limited in the same way. Comparison of motoneurone output to different strength corticospinal inputs and of different motoneurone pools demonstrates an important role for motor unit firing rates in determining the excitability of motoneurones during strong contractions. Next, the reflex actions of group III and IV muscle afferents on motoneurones are investigated. These studies address a long and ongoing debate about the role of these afferents to the slowing of motor unit firing rates during sustained contractions. It was believed that these afferents inhibit motoneurones and contribute to fatigue. However, findings demonstrate that human motoneurones innervating extensor and flexor muscles are not uniformly affected by fatigue-sensitive afferents. Thus afferent inputs from homonymous and antagonist muscles depress extensor motoneurones but facilitate flexor motoneurones. When group III and IV muscle afferents are activated by hypertonic saline, motoneurones of both extensors and flexors are facilitated. This demonstrates parallel excitatory and inhibitory pathways from group III and IV muscle afferents to extensor motoneurones, which are activated under different conditions. Furthermore, the excitation is more pronounced for high-threshold motoneurones. In addition to the effects mediated at motoneurones, activity in group III and IV afferents inhibits motor cortical cells. The final studies investigate changes in the cervical propriospinal pathway with fatigue. This pathway transmits part of the voluntary drive to motoneurones, in parallel with the direct corticospinal pathway. The studies demonstrate that during fatigue, there are coordinated changes in the excitation mediated via this pathway to motoneurones of both fatigued and non-fatigued muscles of the upper limb. In summary, this thesis demonstrates novel aspects of the corticospinal control of motoneurones during voluntary contraction and fatigue.

Amyotrophic Lateral Sclerosis (ALS) is a fatal disease characterized by the progressive degeneration of motor neurons. Although there has been some progress in the identification of genes linked to inherited cases of ALS, the etiology of this disease remains largely unknown. Clinical progression of motor neuron diseases is associated with the degeneration of the axon preceding cell death. Elucidating novel mechanisms important for motor axon maintenance will help gain greater insight into disease pathogenesis. Here, I report that mutations in ras-opposite (rop), which encodes the Drosophila homologue of mammalian Sec1/Munc18, cause progressive degeneration of motor axons while sensory axons are largely unaffected. While mutations in mammalian munc18-1 have been linked to degeneration of the spinal cord, the mechanisms by which this occurs are unknown. Using Drosophila, I found that RNAi-induced knockdown of rop leads to severe motor deficits in adult flies. In addition, I discovered that motor axon degeneration in rop mutants could be delayed by overexpression of the neuronal maintenance factor Nmnat. Interestingly, I found that Rop is localized with Nmnat at the neuromuscular junction and that Rop physically interacts with Nmnat in vivo. These data indicate a novel role for Rop in motor axon maintenance and provide insight into the pathogenesis of neurodegenerative diseases targeting motor neurons, such as ALS.

Thesis (Ph. D.)--University of Hong Kong, 2001. / Includes bibliographical references (leaves 123-155).

Many recent studies on the segmental motor control system have employed spike-triggered-averaging (STA) and other forms of cross-correlation to either attribute CNS, reflex, or direct motor effects to the impulses of a single (reference) neuronal spike train or to explore conditions under which pairs of neural units show temporal correlations in their discharge. Our experience with these techniques suggested the need for a control procedure that tests for synchrony between the reference and other spike trains such as to: (1) either preclude that the observed effects are due to spike trains other than or in addition to the reference train; or (2) give insight into the conditions leading to correlated discharge between two units. A motor unit synchronization test based on analysis of EMG waveforms has already been described. We have modified this test for the detection of synchrony between either afferent or efferent signals by analysis of averaged muscle nerve signals rather than EMG waveforms. Our procedure involves use of a multi-unit muscle nerve recording that serves as the input to a signal averager triggered by a spike train from either: (1) a motor unit's EMG; (2) a dorsal root filament or ganglion cell; or (3) a ramdom trigger source. With appropriate delay of the muscle nerve signal input, the non-rectified average of the trigger signal's waveform is compared to the rectified average which contains this waveform together with contributions of all other active unitary events. Additionally, the rectified average is compared to a "randomly" triggered average of the same input signal. On the basis of these recordings, it can be determined, within certain boundary conditions, whether or not any other unitary events are in synchrony with the reference event. Such synchronization is expressed quantitatively in the form of a synchronization index (SI). We evaluated the efficacy of the SI by electronic simulation procedures and by comparing its use to that of a cross-correlation procedure that tests for synchrony on the basis of crosscorrelograms computed between two simultaneously recorded spindle afferent spike trains during brief stretch of a passive muscle at progressively increasing amplitudes (5 - 100um). These experiments revealed that the SI is a sensitive test of afferent synchrony in the passive muscle provided the spike trains of interest have a signal-to-noise (S/N) ratio > 0.2 in the muscle nerve recording and that it is recognized that the detectable degree of synchronization of a non-reference event is a function of its S/N ratio. For tests on the active muscle, the force levels must remain low. Otherwise increased neuronal activity in the muscle nerve recording decreases the S/N ratio of individual spike trains. Thus, despite restrictive (but predictable) boundary conditions, the SI test can contribute importantly to select conclusions drawn from cross-correlation studies.

No description available.