The human primary motor cortex (M1) undergoes structural and functional change throughout life by a process known as neuroplasticity. Techniques which artificially induce neuroplastic changes are seen as potential adjunct therapies for neurological conditions reliant on neuroplasticity for recovery of function. Unfortunately, the reported improvements in function when these techniques have been used in combination with regular rehabilitation have so far been inconsistent. One reason attributed to this is the large variability in effectiveness of these techniques in inducing neuroplastic change. This thesis has investigated factors influencing the effectiveness and reproducibility of neuroplasticity induction in human M1 using several experimental paradigms. The effectiveness and reproducibility of inducing neuroplasticity in human M1 using two variants of a paired associative stimulation (PAS) protocol was investigated in the first set of experiments (Chapter 2). Both protocols repeatedly paired a peripheral electrical stimulus to the median nerve of the left wrist with single-pulse transcranial magnetic stimulation (TMS) delivered 25 ms later to the contralateral M1. Neuroplastic changes were quantified by comparing the amplitude of the muscle evoked potential (MEP) recorded in abductor pollicis brevis (APB) muscle by suprathreshold TMS prior to and following PAS. With both protocols, neuroplasticity induction was more effective, and the responses across sessions more reproducible, if the experiments were performed in the afternoon compared to the morning. Subsequent experiments confirmed the time of day modulation of PAS-induced neuroplasticity by repeatedly testing twenty-five subjects on two separate occasions, once in the morning (8 am), and once in the evening (8 pm) (Chapter 3). Time of day was also shown to modulate GABAergic inhibition in M1. In a further set of experiments, a double-blind, placebo-controlled study demonstrated that artificially elevated circulating cortisol levels (with a single oral dose of hydrocortisone) inhibits PAS-induced neuroplasticity in the evening (8 pm), indicating that the time of day modulation of neuroplasticity induction with PAS is due, at least in part, to differences in circulating cortisol levels (Chapter 3). The cortical circuits that are modulated by PAS have also been shown to be important in motor learning. Therefore, the final set of experiments, described in Chapter 4, investigated whether motor-training-related changes in motor performance (and cortical excitability) following a ballistic motor training task are also modulated by time of day. Twenty-two subjects repeatedly abducted their left thumb with maximal acceleration for thirty minutes during two experimental sessions (morning (8 am) and evening (8 pm)) on separate occasions. Motor training improved motor performance, and increased cortical excitability, however these changes were independent of time of day. It may be that the motor training task and/or outcome measures used were not sufficiently sensitive to detect a subtle time of day effect of motor training on motor performance. Alternatively, the normally functioning motor system may be able to compensate for changes in cortical excitability to maintain optimal motor performance. These findings have important implications for therapies reliant on neuroplasticity for recovery of function, and indicate that rehabilitation may be most effective when circulating cortisol levels are low. / Thesis (Ph.D.) - University of Adelaide, School of Molecular and Biomedical Science, 2009

The human primary motor cortex (M1) undergoes structural and functional change throughout life by a process known as neuroplasticity. Techniques which artificially induce neuroplastic changes are seen as potential adjunct therapies for neurological conditions reliant on neuroplasticity for recovery of function. Unfortunately, the reported improvements in function when these techniques have been used in combination with regular rehabilitation have so far been inconsistent. One reason attributed to this is the large variability in effectiveness of these techniques in inducing neuroplastic change. This thesis has investigated factors influencing the effectiveness and reproducibility of neuroplasticity induction in human M1 using several experimental paradigms. The effectiveness and reproducibility of inducing neuroplasticity in human M1 using two variants of a paired associative stimulation (PAS) protocol was investigated in the first set of experiments (Chapter 2). Both protocols repeatedly paired a peripheral electrical stimulus to the median nerve of the left wrist with single-pulse transcranial magnetic stimulation (TMS) delivered 25 ms later to the contralateral M1. Neuroplastic changes were quantified by comparing the amplitude of the muscle evoked potential (MEP) recorded in abductor pollicis brevis (APB) muscle by suprathreshold TMS prior to and following PAS. With both protocols, neuroplasticity induction was more effective, and the responses across sessions more reproducible, if the experiments were performed in the afternoon compared to the morning. Subsequent experiments confirmed the time of day modulation of PAS-induced neuroplasticity by repeatedly testing twenty-five subjects on two separate occasions, once in the morning (8 am), and once in the evening (8 pm) (Chapter 3). Time of day was also shown to modulate GABAergic inhibition in M1. In a further set of experiments, a double-blind, placebo-controlled study demonstrated that artificially elevated circulating cortisol levels (with a single oral dose of hydrocortisone) inhibits PAS-induced neuroplasticity in the evening (8 pm), indicating that the time of day modulation of neuroplasticity induction with PAS is due, at least in part, to differences in circulating cortisol levels (Chapter 3). The cortical circuits that are modulated by PAS have also been shown to be important in motor learning. Therefore, the final set of experiments, described in Chapter 4, investigated whether motor-training-related changes in motor performance (and cortical excitability) following a ballistic motor training task are also modulated by time of day. Twenty-two subjects repeatedly abducted their left thumb with maximal acceleration for thirty minutes during two experimental sessions (morning (8 am) and evening (8 pm)) on separate occasions. Motor training improved motor performance, and increased cortical excitability, however these changes were independent of time of day. It may be that the motor training task and/or outcome measures used were not sufficiently sensitive to detect a subtle time of day effect of motor training on motor performance. Alternatively, the normally functioning motor system may be able to compensate for changes in cortical excitability to maintain optimal motor performance. These findings have important implications for therapies reliant on neuroplasticity for recovery of function, and indicate that rehabilitation may be most effective when circulating cortisol levels are low. / Thesis (Ph.D.) - University of Adelaide, School of Molecular and Biomedical Science, 2009

A Neuronavegação é uma técnica de visualização computacional da localização de instrumentos em relação às estruturas neuronais. A estimulação magnética transcraniana (EMT) é uma ferramenta para estimulação cerebral não-invasiva, que tem sido utilizada em aplicações clínicas, para o tratamento de algumas patologias, e também em pesquisas. Entretanto, a EMT é uma técnica altamente dependente de parâmetros como o posicionamento e orientação da bobina de estimulação em relação às estruturas neuronais. Para auxiliar no posicionamento da bobina, uma combinação entre neuronavegação e EMT é utilizada, chamada de EMTnavegada (EMTn). Essa técnica permite o monitoramento em tempo real da bobina de EMT em relação às neuroimagens. Porém, a utilização da EMTn ainda é pouco explorada, tanto na pesquisa quanto no ambiente clínico, devido ao alto custo, exigência da imagem de ressonância magnética, complexidade e baixa portabilidade dos sistemas de EMTn comerciais. O neuronavegador de código aberto e livre, InVesalius Navigator, vem sendo desenvolvido para ajudar a suprir essa necessidade. Assim, o objetivo desta dissertação foi desenvolver ferramentas para o sistema de neuronavegação InVesalius Navigator. As funcionalidades adicionadas foram: i) suporte para três tipos de rastreadores espaciais; ii) sincronização da EMT com o neuronavegador; iii) guia para o reposicionamento da bobina. Além disso, com intuito de contornar a necessidade de utilizar a imagem de ressonância magnética foram realizados estudos para a substituição por uma imagem padrão. Na parte de desenvolvimento, experimentos de caracterização foram realizados para validação das ferramentas. O sistema de neuronavegação apresentou-se intuitivo e de fácil portabilidade. Além disso, a precisão obtida foi semelhante à de sistemas comerciais. Os erros de localização foram inferiores a 3 mm, considerados aceitáveis para aplicações clínicas. Na segunda parte, procedimentos que não exigem extrema precisão, como a localização e digitalização do hotspot, a variabilidade foi considerada aceitável. Portanto, a utilização da imagem média mostrou-se uma possível alternativa para as imagens de ressonância magnética. / Neuronavigation is a computer image-guided technique to locate surgical instruments related to brain structures. The transcranial magnetic stimulation (TMS) is a non-invasive brain stimulation method, it has been used for clinical purposes, treating neurological disorders, and also for research purpose, studying cortical brain function. However, the use of TMS is highly dependent on coil position and orientation related to brain structures. The navigated TMS (nTMS) is a combined technique of neuronavigation system and TMS, this technique allows tracking TMS coil by image guidance. Yet, nTMS is not widely used, either in research and in the clinical environment, due to the high cost, magnetic resonance imaging requirement, complexity, and low portability of commercial TMS systems. Thus, the aim of this dissertation was to develop tools for the neuronavigator system InVesalius Navigator, such as: i) support for three types of spatial trackers; ii) synchronization of the TMS with the neuronavigator; iii) guide for coil repositioning. In addition, in order to overcome the magnetic resonance imaging requirement, studies were made to replace it with a standard brain image. In the development part, characterization experiments were done to validate the new functionalities. Therefore, the accuracy obtained was similar to commercial systems. Localization errors were less than 3 mm considered acceptable for clinical applications. In the second part, for procedures that do not require extreme accuracy, such as the location and scanning of the hotspot, the variability was considered acceptable. Therefore, the use of the standard brain image was a possible alternative for magnetic resonance imaging.

Fatigue is defined as an exercise-induced decrease in maximal voluntary force produced by a muscle. Fatigue may arise from central and/or peripheral mechanisms. Supraspinal fatigue (a component of central fatigue) is defined as a suboptimal output from the motor cortex and measured using transcranial magnetic stimulation (TMS). Reductions in O2 supply (hypoxia) exacerbate fatigue and as the severity of hypoxia increases, central mechanisms of fatigue are thought to contribute more to exercise intolerance. In study 1, the feasibility of TMS to measure cortical voluntary activation and supraspinal fatigue of human knee-extensors was determined. TMS produced reliable measurements of cortical voluntary activation within- and between-days, and enabled the assessment of supraspinal fatigue. In study 2, the mechanisms of fatigue during single-limb exercise in normoxia (arterial O2 saturation [SaO2] ~98%), and mild to severe hypoxia (SaO2 93-80%) were determined. Hypoxia did not alter neuromuscular function or cortical voluntary activation of the knee-extensors at rest, despite large reductions in cerebral oxygenation. Maximal force declined by ~30% after single-limb exercise in all conditions, despite reduced exercise time in severe-hypoxia compared to normoxia (15.9 ± 5.4 vs. 24.7 ± 5.5 min; p < 0.05). Peripheral mechanisms of fatigue contributed more to the reduction in force generating capacity of the knee-extensors following single-limb exercise in normoxia and mild- to moderate-hypoxia, whereas supraspinal fatigue played a greater role in severe-hypoxia. In study 3, the effect of constant-load cycling exercise to the limit of tolerance in hypoxia (SaO2 ~80%) and normoxia was investigated. Time to the limit of tolerance was significantly shorter in hypoxia compared to normoxia (3.6 ± 1.3 vs. 8.1 ± 2.9 min; p < 0.001). The reductions in maximal voluntary force and knee-extensor twitch force at task-failure were not different in hypoxia compared to normoxia. However, the level of supraspinal fatigue was exacerbated in hypoxia, and occurred in parallel with reductions in cerebral oxygenation and O2 delivery. Supraspinal fatigue contributes to the decrease in whole-body exercise tolerance in hypoxia, presumably as a consequence of inadequate O2 delivery to the brain.

L'empathie nous permet de comprendre et de réagir aux sensations des autres individus. Regarder une situation douloureuse peut induire des comportements de type prosociaux orientés vers les autres ou des réponses d'évitement comme celles enregistrées en réponse à une menace. Le but principal de cette thèse était d'étudier les comportements d'approche/évitement et freezing qui se produisent soit en observant la douleur des autres, soit pendant l'anticipation de la douleur. Deux tâches manipulant la prise de perspective ont permis d'enregistrer des cotations supérieures concernant le niveau de douleur, des temps de réaction inférieurs (expérience 1) et des index de réponses d'évitement plus grands (expérience 2) pour la perspective consistant à imaginer que le sujet représenté dans une condition douloureuse était la personne la plus aimée. Dans la troisième expérience, nous avons analysé le comportement du freezing au niveau du système corticospinal du participant : un effet du freezing spécifique fut rapporté uniquement lorsque de la présentation des stimuli douloureux en perspective du première personne. Dans une quatrième expérience, l'effet du freezing, normalement présent en réponse aux stimuli douloureux fut aussi rapporté dans le cadre de l'anticipation de la douleur pour soi-même. Nos études suggèrent que ce sont principalement les mécanismes cognitifs de prise de perspective qui modulent la réponse empathique et que la perspective de la personne la plus aimée induise la réponse empathique la plus forte. Au contraire les réponses du freezing des modulations corticospinales sont principalement observées lorsque le sujet adopte une perspective en première personne / Empathy allows us to understand and react to other people feelings. Regarding empathy for pain, a witness looking at a painful situation may react to other-oriented and prosocial-altruistic behaviors or self-oriented withdrawal responses. The main aim of this thesis was to study approach/avoidance and freezing behavioral manifestations that co-occurring along with both others’ pain observation and during the anticipation of pain. In two perspective-taking tasks, we investigated the influence of the type of relationship between the witness and the target in pain. Results showed that higher pain ratings, lower reactions times (experiment 1) and greater withdrawal avoidance postural responses (experiment 2) were attributed when participants adopted their most loved person perspective. In experiment 3, we analyzed the freezing behavior in the observer’s corticospinal system while subject was observing painful stimuli in first-and third-person perspectives. Results showed the pain-specific freezing effect only pertained to the first-person perspective condition. An empathy for pain interpretation suggests empathy might represent the anticipation of painful stimulation in oneself. In experiment 4 results, we found that the freezing effect present during a painful electrical stimulation was also present in the anticipation of pain. In conclusion, our studies suggest that cognitive perspective-taking mechanisms mainly modulate the empathic response and the most loved person perspective seems to be prevalent. In addition, more basic pain-specific corticospinal modulations are mainly present in the first-person perspective and it seems to not be referred to the empathy components

Transcranial magnetic stimulation (TMS) is a non-invasive technique that can be used to assess the integrity neuronal circuits in the motor cortex, both at the intrahemispheric and interhemispheric level. In the present study, TMS was used to examine age-related modulation of corticospinal function. Participants underwent hand function testing to examine possible links between TMS measures and manual ability. Participants consisted of healthy young (n=13) and senior (n=17) right-handed individuals. Hand function testing consisted of a battery of tests administered bilaterally to assess each participant’s dexterity, strength, movement speed and reaction time. The following TMS measures were assessed bilaterally: resting motor threshold, recruitment curve and silent periods of the contralateral and ipsilateral hand. Both young and senior subjects showed significant intermanual differences in most behavioral measures, favoring their dominant right hand. There was an age-related difference in TMS measures indicating a decline in intrahemispheric excitability and interhemispheric inhibition. A general trend linking specific TMS measures in the active state with age-related changes in hand function on the dominant hand was found. Our results suggest that TMS markers of corticospinal excitability can be used to predict declining hand function with age and thus could provide an early diagnosis of pathological aging.

Theta-burst stimulation (TBS) is a form of repetitive transcranial magnetic stimulation (TMS) developed to induce neuroplasticity. TBS usually consists of 50 Hz bursts at 5 Hz intervals. When applied intermittently, it can lead to facilitation of motor evoked potentials (MEPs), although these effects can be variable between individuals. Here, we aimed to determine whether a version of intermittent TBS (iTBS) consisting of 30 Hz bursts at 6 Hz intervals would produce less variable modulation. Nineteen healthy adults underwent single-pulse TMS to assess corticomotor excitability at baseline as reflected in MEP amplitude. 30 Hz iTBS was then administered and MEP amplitude was reassessed at 5-, 20- and 45-mins after the iTBS protocol. Compared to baseline, MEPs were significantly facilitated up to 45-min post-iTBS and most participants exhibited the expected facilitation. These observations suggest that 30 Hz/6 Hz iTBS may provide a sound alternative to induce consistent neuromodulatory effects over the commonly used 50 Hz/5 Hz protocol.

Blindsight is a phenomenon in which patients suffering damage to the primary visual cortex (V1) perceive themselves as blind, but nonetheless seem to have some residual capacity to distinguish between visual stimuli better than chance. Blindsight can be divided into two subtypes: blindsight type I and blindsight type II. Blindsight type I is defined as visual capacity in the absence of acknowledged awareness. Patients with blindsight type II have visual capacity with some feeling or sensation in the blind field. Visual pathways bypassing V1 are assumed to be responsible for the residual capacity in blindsight. To investigate whether these pathways are present in healthy individuals we examined if it is possible to induce blindsight in healthy individuals by reviewing studies that have tried to induce blindsight with transcranial magnetic stimulation (TMS). We found that TMS might be able to induce blindsight type I of side detection. We also found that TMS might be able to induce blindsight type II of colour, orientation, and trustworthiness. Further, we found that both conscious and unconscious perception of shapes are dependent on processing in early visual cortex (EVC) in healthy individuals. We conclude that the full capacity seen in blindsight is most probable caused by neural reorganisation post trauma. The visual pathways bypassing V1 are, if present in healthy individuals, too weak to influence behaviour with the possible exception of side detection. Additionally, we conclude that the use of a binary awareness scale in blindsight studies fails to capture vaguely seen stimuli.

No / Until recently, it was widely believed that object position and object motion were represented independently in the visual cortex. However, several studies have shown that adaptation to motion produces substantial shifts in the perceived position of subsequently viewed stationary objects [[13]]. Two stages of motion adaptation have been proposed: an initial stage at the level of V1 and a secondary stage thought to be located in V5/MT [[4]]. Indeed, selective adaptation can be demonstrated at each of these levels of motion analysis [[5, 6]]. What remains unknown is which of these cortical sites are involved in modulating the positional representation of subsequently viewed objects. To answer this question directly, we disrupted cortical activity by using transcranial magnetic stimulation (TMS) immediately after motion adaptation. When TMS was delivered to V5/MT after motion adaptation, the perceived offset of the test stimulus was greatly reduced. In marked contrast, TMS of V1 had no effect on the changes that normally occur in perceived position after motion adaptation. This result demonstrates that the anatomical locus at which motion and positional information interact is area V5/MT rather than V1/V2.

Yes / Human neuropsychological and neuroimaging studies have raised the possibility that different attributes of optic flow stimuli, namely radial direction and the position of the focus of expansion (FOE), are processed within separate cortical areas. In the human brain, visual areas V5/MT+ and V3A have been proposed as integral to the analysis of these different attributes of optic flow stimuli. In order to establish direct causal relationships between neural activity in V5/MT+ and V3A and the perception of radial motion direction and FOE position, we used Transcranial Magnetic Stimulation (TMS) to disrupt cortical activity in these areas whilst participants performed behavioural tasks dependent on these different aspects of optic flow stimuli. The cortical regions of interest were identified in seven human participants using standard fMRI retinotopic mapping techniques and functional localisers. TMS to area V3A was found to disrupt FOE positional judgements, but not radial direction discrimination, whilst the application of TMS to an anterior sub-division of hV5/MT+, MST/TO-2, produced the reverse effects, disrupting radial direction discrimination but eliciting no effect on the FOE positional judgement task. This double dissociation demonstrates that FOE position and radial direction of optic flow stimuli are signalled independently by neural activity in areas hV5/MT+ and V3A. / This work was funded by the BBSRC (grant B/N003012/1).