L-glutamate (Glu) is the predominant excitatory neurotransmitter inthe mammalian central nervous system (CNS) and is associated with a widevariety of functions including motor behavior and sensory perception. Whilemicrodialysis methods have been used to record tonic levels of Glu, little isknown about the more rapid changes in Glu signals that may occur in awakeanimals. We have previously reported acute recording methods using anenzyme-based microelectrode array (MEA) with fast temporal resolution (800msec), that is minimally invasive and is capable of detecting low levels of Glu (andlt;0.2 ??M) in anesthetized animals with little interference from other analytes. Wehave made a series of modifications to the MEA design to allow for reliablemeasures in the brain of awake behaving rats. In these studies, wecharacterized the effects of chronic implantation of the MEA into the striatum andprefrontal cortex (PFC) of Fischer 344 and Long Evans rats. We measuredresting levels of Glu and local application of Glu for 7 days without a significantloss of sensitivity and determined that Glu measures due to exogenous Gluvaried between rat strain and brain region. In addition, we determined theviability of the recordings in the brains of awake animals. We performed studiesof tail-pinch induced stress which caused an increase in Glu in the striatum andPFC of Long Evans and Fischer 344 rats. Histological data show that chronicimplantation of our MEAs caused minimal injury to the CNS. Taken together, ourdata support that chronic recordings of tonic and phasic Glu can be carried out inawake rats reliably for 7 days in vivo allowing for longer term studies of Gluregulation in behaving rats.

<p>In recent years, large-scale brain recordings in nonhuman primates have been a driving force for both fundamental neuroscience and the field of brain-machine interfaces (BMIs). This required monkey implants connected to external amplifiers and computers with ever increasing number of cables. As shown with our recent demonstration of 2,000 neurons recorded in one monkey, a tethered recording system begins to get bulky and complex, particularly for our BMI and neurophysiological research. To address this problem, we developed a multichannel wireless recording framework. The system was been tested in freely moving rhesus monkey by integrating wireless neural recordings with external computers performing BMI decoding, behavioral manipulanda and optical tracking. This technology can be applied to primate behavior research and, in the near future, wireless, fully implantable human neuroprosthetics, which is of great significance to those suffering from locomotor deficiencies, such as those brought on by spinal cord injury and stroke. Aided with these advances, I was able to study monkeys in unrestrained locomotion while their cortical activity was continuously monitored. I also explored unrestrained behaviors and how they showed distinct transitions in neural dynamics as monkeys engaged in different behavioral activities or learned new motor skills, such as bipedal walking. I was able to decode them many of these behavioral states from cortical activity with neural classifiers. Lastly, monkeys were able to perform BMI tasks continuously for many hours, allowing us to prove the relevance of unrestrained noise in BMI performance. Lastly, I present my role in developing two brain actuated movement platforms, a robotic exoskeleton under the guise of the WalkAgain project, and a microelectrode BMI enabled wheelchair. This body of work should assist those on the path to the next generation of clinical neuroprostheses and neural communication systems.</p> / Dissertation

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Animals must sense their surroundings to update their internal representations of the external environment, and exploratory behaviours such as sensory sampling are influenced by past experiences. This thesis investigates how voluntary sensory sampling activities undergo learning-dependent changes. Studies of freely behaving animals impose two major challenges: 1) the accuracy of biological measurements is compromised by movement-induced artifacts, and 2) large degrees of freedom in unrestrained behaviours confound well-controlled studies. Pulse-type weakly electric fish (WEF) are an ideal choice to study adaptive sensory sampling from unrestrained animals, since they generate readily observable and quantifiable sensory capture events expressed by discrete pulses of electric organ discharges (EODs). To study the voluntarily movements and sensory sampling while animals navigated in darkness, we developed three novel experimental techniques to track movements and detect sensory sampling from a freely behaving WEF: 1) an EOD detector to remotely and accurately measure the sensory sampling rate, 2) an electrical tracking method to track multiple WEF using their own EODs, and 3) visual tracking algorithm for robust body tracking through water under infrared illumination. These techniques were successfully applied to reveal novel sensory sampling behaviours in freely exploring Gymnotus sp. Cortical activity precedes self-initiated movements by several seconds in mammals; this observation has led into inquiries on the nature of volition. Here we demonstrate the sensory sampling enhancement also precedes self-initiated movement by a few seconds in Gymnotus sp. Next, we tested whether these animals can be trained to learn a location of food using electrically detectable landmarks and, if so, whether they can use their past experiences to optimize their sensory sampling. We found that animals revisited the missing food location with high spatial accuracy, and they intensified their sensory sampling near the expected food location by increasing the number of EOD pulses per unit distance travelled.

Les techniques optogénétiques ont laissé entrevoir un potentiel exceptionnel dans l’étude des mécanismes gouvernant l’intégration de l’information dans le système nerveux. Afin d’établir les relations existant entre des séquences d’activité neuronale définies et le comportement, les techniques optiques doivent permettre d’appréhender des groupes de neurones avec une résolution cellulaire chez l’animal éveillé et mobile. Jusqu’alors, l’activité neuronale chez l’animal vigile non contraint n’a été contrôlée que via l’illumination en champs large. Dans ce travail, nous démontrons une résolution proche de la cellule unique pour la photoactivation, chez l’animal éveillé et mobile, à l’aide d’un fibroscope. Les motifs de photoactivation, produits par holographie, sont transmis jusqu’à la souris par un guide d’image couplé à un micro-objectif. Une imagerie de fluorescence permet de localiser les cellules d’intérêt et d’enregistrer l’activité neuronale, par épifluorescence, illumination structurée, ou encore microscopie confocale multi-point sans balayage. Le fibroscope est testé chez l’animal anesthésié et chez l’animal éveillé et mobile, dont les interneurones de la couche moléculaire du cervelet expriment les protéines ChR2-tdTomato et GCaMP. Nous avons généré des signaux calciques somatiques en ciblant les corps cellulaires avec des spots de photoactivation de 5µm de diamètre. Chez l’animal anesthésié, nous avons démontré que la photoactivation pouvait être réalisée avec une résolution latérale de 10µm et une résolution axiale de 40µm, en considérant la demi-largeur à mi-hauteur de la courbe de résolution. Nous avons montré qu’un ou plusieurs soma pouvaient être ciblés sélectivement. Chez l’animal éveillé et mobile, le champs de vue est resté stable au cours des acquisitions. Nous avons trouvé une résolution latérale pour la photoactivation égale à 10µm, démontrant une résolution de photoactivation proche de la cellule unique chez l’animal vigile non contraint. / Optogenetics has shown great potential to study the mechanisms governing information integration in the brain. To link specific spatiotemporal activity patterns and behaviours, optical methods should provide simultaneous access to a group of neurons with single cell resolution in freely behaving animals. So far, however, optogenetic control of neural activity in freely behaving rodents has been performed with widefield illumination only. Here, we demonstrate photoactivation with near-cellular resolution in freely behaving mice using a fiberscope. Photoactivation patterns, produced with computer-generated holography, were transmitted to the mouse using a fiber bundle coupled to a micro-objective. Fluorescence imaging allowed locating cells and recording neuronal activity, via either epifluorescence, structured illumination, or scanless multi-point confocal microscopy. The fiberscope was tested both in anesthetized and freely-behaving mice co-expressing ChR2-tdTomato and GCaMP proteins in cerebellar molecular layer interneurons. By targeting an interneuron soma with a 5µm diameter photoactivation spot, we could elicit a calcium transient. In anesthetized animals, we demonstrated that photoactivation could be performed with 10µm and 40µm lateral and axial resolution, half-width at half maximum, respectively. We showed that either a single or multiple somata could be selectively targeted. In awake unrestrained animals, the field of view remained stable during our acquisitions. We found that photoactivation lateral resolution remained equal to 10µm, demonstrating photoactivation with near-cellular resolution in freely behaving mice.

This research builds on previous studies that have documented evidence of Professor Anna Craft’s concept of ‘Possibility Thinking’ (PT) as at the heart of creativity which involves children transitioning from ‘what is this?’ to ‘what can I or we do with this?’ as well as imagining ‘as if’ they were in a different role. My thesis titled “Communicating Possibilities” examines English nursery children's emergent creativity, exploring the three to four-year-old child as an artistic communicator and possibility thinker through a case study approach situated in one primary school in South West England. Three main research questions were posed concerning the ‘what, how, and why’ of creativity when children communicated through art; as well as exploring the nurturing role of others, and identity manifest through voice and learning experience. This doctoral study is essentially interpretivist in nature seeking to explain how people make sense of their social worlds, and is an exploration framed by culturally negotiated, shared meanings, and complex social relations. Data was collected over one school year, in three nine-week research phases by the following ethnographic methods: naturalistic observations; researcher diary; children’s creative journals; and practitioner interviews. These methods were repeated for each phase. Inductive and deductive data analysis was conducted. Undertaken over time as the project unfolded, a grounded theory approach was applied in total to 27 episodes. Micro event analysis of creative behaviours in action and narrative discourses of two kinds: peer-to-peer, and child-to-adult (teacher, early years practitioner, and my researcher dialogue) revealed four broad critical themes: Observing and documenting children’s creativity; What children can do together- recognising differences; Pedagogy of possibilities- developing a role; and The value of artistic communication in the nursery classroom. Each is discussed in terms of the key implications these themes hold for theory, policy, and early years practice.

Der primäre somatosensorische Kortex (S1) besteht aus sechs Schichten (L1L6).Die koordinierte Aktivität dieser sechs Schichten kortikaler Neurone ist entscheidend für die sensorische Wahrnehmung und die Steuerung willkürlichen Verhaltens. Es ist jedoch noch wenig über die synaptischen Mechanismen bekannt, die die Verarbeitung zwischen den kortikalen Schichten bei sich aktiv verhaltenden Tieren bestimmen. Ich habe einfache und doppelte in vivoGanzzellableitungen im VorderpfotenAreal von S1 in der Maus gemacht, und gezeigt, dass Pyramidalzellen in L2/3 und L5 während einer Bewegung der Vorderpfote Unterschiede in ihren intrinsischen Eigenschaften und der Dynamik ihrer Membranpotenziale zeigen. Doppelableitungen haben gezeigt, dass sensorisch und motorisch ausgelöste synaptische Eingänge zwischen den Zellschichten weitgehend korreliert waren, niederfrequente unterschwellige Potenzialschwankungen und spontane Aktionspotenziale jedoch einen schichtspezifischen Zeitverlauf zeigten. Auf einer längeren Zeitskala beobachteten wir, dass spontane Bewegungen der Vorderpfote eine Dekorrelation unterschwelliger Aktivität zwischen den Schichten auslösten. Des Weiteren zeigten L5Pyramidalzellen durch ihre Aktivität sensorisch ausgelöste und spontane Bewegungen der Vorderpfote stärker an, als L2/3Neurone. Insgesamt deuten meine Daten darauf hin, dass Unterschiede zwischen den Zellschichten beim Timing von Aktionspotenzialen, bei der unterschwelligen Synchronisierung und bei den mittleren Feuerraten sowohl von der Quelle des zu Grunde liegenden synaptischen Eingangs als auch vom resultierenden Verhalten abhängen. Außerdem konnte ich zeigen, dass Neurone im VorderpfotenAreal von S1 auf leichte Kältereizung der Vorderpfote antworten, und dass diese Antwort vom Ionenkanal transient receptor potential cation channel subfamily M member 8 (TRPM8) in primären sensorischen afferenten Neuronen vermittelt wird. / The primary somatosensory cortex (SI) is composed of six layers (L1L6). The coordination of neural activities across six layers of cortical neurons is essential for reliable sensory perception and the control of voluntary behavior. However, the synaptic neural mechanisms governing translaminar cortical processing in behaving animals are still unknown. I made in vivo single and dual whole cell recordings in mouse forepaw SI, my work revealed that L2/3 and L5 pyramidal neurons have distinct intrinsic properties and membrane potential dynamics during forepaw behavior. Dual recordings showed that sensory and movement evoked synaptic inputs were closely correlated across layers, but low frequency subthreshold fluctuations and spontaneous action potentials exhibited a laminar specific temporal profile. At longer time scales, my data showed that spontaneous forepaw movement evoked a decorrelation of subthreshold activity across layers. Furthermore, L5 pyramidal neurons signaled sensory evoked and spontaneous forepaw movements more strangely than L2/3 neurons. Overall, my work suggests that laminar differences in the timing of action potential firing, subthreshold synchrony and mean firing rates are dependent both on the origin of the underlying synaptic input and the behavioral outcome of the event. In addition, I identified that forepaw SI neurons respond to mild cooling stimulation of the forepaw and that this response is mediated by the Transient receptor potential cation channel subfamily M member 8 (TRPM8) in primary sensory afferent neurons.

The following diploma thesis deals with the analysis of two short stories by Adalbert Stifter from his collection Colorful stones - Rock Crystal and Granit. They were deliberately chosen, because in both stories children are the main protagonists. In these short stories there is always a boy and a girl, while the boy is always the one more experienced, and he protects the girl and brings her to safety. She follows him and obeys his instructions. Although the text contains many descriptions, it does not act statically because the heroes are on the run from danger, which gives the action dynamics. The text, the story, the language and the plot of the short stories as well as their common elements are analyzed. Furthermore, the theoretical concept, which is formulated in the preface to this collection, and whose major concepts are big and small, is confronted with the realization in these short stories. KEYWORDS Life, experience, ordinary, extraordinary, child, behaving, love, danger, nature, landscape, death