Electroretinoraphic signals (ERGs) are affected when recorded under isoflurane anesthesia in the operating room. We explored the effect of isoflurane and propofol in ex vivo guinea pig retinal preparations using a multielectrode array to record simultaneously ERGs and retinal ganglion cell (RGC) activity. The viability and light-response characteristics of the model were documented. In the presence of isoflurane, the ERG and RGC activity was reduced in a dose-dependent manner, even at sub-clinical doses; the OFF responses were consistently more affected. Propofol had minimal effects: at subclinical doses, a small excitation was measured while a concentration a hundred times stronger than the clinical concentration was required to measure a significant decline in EGR and RGC signals. This study confirms the usefulness of the guinea pig model to study clinically relevant retinal issues and shows that propofol is a better anesthetic to use in the operating room when retinal investigations are required.

Yes / Sonic Hedgehog (SHH) is a glycoprotein associated with development that is also expressed in the adult CNS and released after brain injury. Since the SHH receptors PTCH1 (patched homolog-1) and SMO (Smoothened) are highly expressed on astrocytes, we hypothesised that SHH regulates astrocyte function. Primary mouse cortical astrocytes derived from embryonic (E15) Swiss mouse cortices, were treated with two chemically distinct agonists of the SHH pathway, which caused astrocytes to elongate and proliferate. These changes are accompanied by decreases in the major astrocyte glutamate transporter, GLT-1 and the astrocyte intermediate filament protein GFAP. Multi-site electrophysiological recordings revealed that the SHH agonist, SAG supressed neuronal firing in astrocyte-neuron co-cultures and this was abolished by the astrocyte metabolic inhibitor ethylfluoroacetate, revealing that SHH stimulation of metabolically-active astrocytes influences neuronal firing. Using 3D co-culture, MAP2 western blotting and immunohistochemistry, we show that SHH-stimulated astrocytes protect neurons from kainate induced cell death. Altogether the results show that SHH regulation of astrocyte function represents an endogenous neuroprotective mechanism. / BBSRC

No description available.

<p>The loss of the ability to walk as the result of neurological injury or disease critically impacts the mobility and everyday lifestyle of millions. The World Heath Organization (WHO) estimates that approximately 1% of the world's population needs the use of a wheelchair to assist their personal mobility. Advances in the field of brain-machine interfaces (BMIs) have recently demonstrated the feasibility of using neuroprosthetics to extract motor information from cortical ensembles for more effective control of upper-limb replacements. However, the promise of BMIs has not yet been brought to bear on the challenge of restoring the ability to walk. A future neuroprosthesis designed to restore walking would need two streams of information flowing between the user's brain and the device. First, the motor control signals would have to be extracted from the brain, allowing the robotic prosthesis to behave in the manner intended by the user. Second, and equally important would be the flow of sensory and proprioceptive information back to the user from the neuroprosthesis. Here, I contribute to the foundation of such a bi-directional brain machine interface for the restoration of walking in a series of experiments in two animal models, designed to show the feasibility of (1) extracting locomotor information from neuronal ensemble activity and (2) sending information back into the brain via cortical microstimulation. </p><p>In a set of experiments designed to investigate the extraction of locomotor parameters, I chronically recorded from ensembles of neurons in primary motor (M1) and primary somatosensory (S1) cortices in two adult female rhesus macaques as they walked bipedally, at various speeds, both forward and backward on a custom treadmill. For these experiments, rhesus monkeys were suitable because of their ability to walk bipedally in a naturalistic manner with training. I demonstrate that the kinematics of bipedal walking in rhesus macaques can be extracted from neuronal ensemble recordings, both offline and in real-time. The activity of hundreds of neurons was processed by a series of linear decoders to extract accurate predictions of leg joints in three dimensional space, as well as leg muscle electromyograms (EMGs). Using a multi-layered switching model allowed us to achieve increased extraction accuracy by segregating different behavioral modes of walking.</p><p>In a second set of experiments designed to investigate the usage of microstimulation as a potential artificial sensory channel, I instructed two adult female Aotus trivirgatus (owl monkeys) about the location of a hidden food reward using a series of cortical microstimulation patterns delivered to primary somatosensory (S1) cortex. The owl monkeys discriminated these microstimulation patterns and used them to guide reaching movements to one of two targets. Here, owl monkeys were used which were previously implanted with electrode arrays of high longevity and stability. These monkeys were previously trained on a somatosensory cued task, which allowed a quick transition to microstimulation cueing. The owl monkeys learned to interpret microstimulation patterns, and their skill and speed of learning new patterns improved over several months. Additionally, neuronal activity recorded on non-stimulated electrodes in motor (M1), premotor (PMD) and posterior parietal (PP) cortices allowed us to examine the immediate neural responses to single biphasic stimulation pulses as well as overall responses to the spatiotemporal pattern. Using this recorded neuronal activity, I showed the efficacy of several linear classification algorithms during microstimulation. </p><p>These results demonstrate that locomotor kinematic parameters can be accurately decoded from the activity of neuronal ensembles, that multichannel microstimulation is a viable information channel for sensorized prosthetics, and that the technical limitations of combining these techniques can be overcome. I propose that bi-directional BMIs integrating these techniques will one day restore the ability to walk to severely paralyzed patients.</p> / Dissertation

This dissertation presents the development of a multielectrode microcompartment platform for understanding signal transduction in the nervous system. The design and fabrication of the system and the characterization of the system for pharmacological and electrophysiological measurements of cultured neurons is presented in this work. The electrophysiological activity of cultured dorsal root ganglion (DRG) neurons and cortical neurons is shown on the MEA substrate. These recordings were measured and tied to the toxicological effects of the chemotherapeutic drug vincristine on DRGs. Conventional electrophysiological recordings (via a patch micropipette) are made routinely to record action potentials and ion channel activity in neurons. Moreover, Campenot chambers (traditional compartmented culture systems) have been used for the last thirty years to study the selective application of drugs to neurons. Both of these techniques are useful and well established; however they have their limitations. For instance, Campenot chambers cannot be used very well for small processs-producing neurons, since the barriers are difficult to tranverse. Moreover, conventional patch recordings are labor-intensive, especially when more than one microelectrode needs to be positioned. The developed system is composed of a two compartment divider, each compartment capable of housing axons or cell bodies. Underneath the divider, the substrate has 60 electrodes, arranged in several lines to accommodate several different neurite tracks. Neurons can be stimulated and their activity can be recorded in both of the compartments. The neurotoxin and chemotherapeutic drug vincristine was tested in the system on the DRGs. The drug caused length-dependent axonal degeneration in the DRGs when applied locally. Moreover, electrophysiological activity in both compartments showed that only the activity in the axonal compartment was affected, leading us to believe that the mechanism behind the degeneration is localized to the distal axon.

No description available.

Willfully controlling the focus of an extracellular stimulus remains a significant challenge in the development of neural prosthetics and therapeutic devices. In part, this challenge is due to the vast set of complex interactions between the electric fields induced by the microelectrodes and the complex morphologies and dynamics of the neural tissue. Overcoming such issues to produce methodologies for targeted neural stimulation requires a system that is capable of (1) delivering precise, localized stimuli a function of the stimulating electrodes and (2) recording the locations and magnitudes of the resulting evoked responses a function of the cell geometry and membrane dynamics. In order to improve stimulus delivery, we developed microfabrication technologies that could specify the electrode geometry and electrical properties. Specifically, we developed a closed-loop electroplating strategy to monitor and control the morphology of surface coatings during deposition, and we implemented pulse-plating techniques as a means to produce robust, resilient microelectrodes that could withstand rigorous handling and harsh environments. In order to evaluate the responses evoked by these stimulating electrodes, we developed microscopy techniques and signal processing algorithms that could automatically identify and evaluate the electrical response of each individual neuron. Finally, by applying this simultaneous stimulation and optical recording system to the study of dissociated cortical cultures in multielectode arrays, we could evaluate the efficacy of excitatory and inhibitory waveforms. Although we found that the proximity of the electrode is a poor predictor of individual neural excitation thresholds, we have shown that it is possible to use inhibitory waveforms to globally reduce excitability in the vicinity of the electrode. Thus, the developed system was able to provide very high resolution insight into the complex set of interactions between the stimulating electrodes and populations of individual neurons.

What changes in our brain when we learn? This is perhaps the most intriguing question of science in this century. In an attempt to learn more about the inner workings of neural circuitry, I studied cultured 2-dimensional networks of neurons on multi-electrode arrays (MEAs). MEAs are ideal tools for studying long-term neural ensemble activity because many individual cells can be studied continuously for months, through electrical stimulation and recording. One of the most prominent patterns of activity observed in these cultures is network-wide spontaneous bursting, during which most of the active electrodes in the culture show elevated firing rates. We view the persistence of spontaneous bursting in vitro as a sign of arrested development due to deafferentation. Substituting distributed electrical stimulation for afferent input transformed the activity in dissociated cultures from bursting to more dispersed spiking, reminiscent of activity in the adult brain. Burst suppression reduced the variability in neural responses making it easier to induce and detect functional plasticity caused by tetanic stimulation. This suggests that spontaneous bursts interfere with the effects of external stimulation and that a burst-free environment leads to more stable connections and predictable effects of tetanization. Moreover, our culture models continuously receive input stimulation in the form of background electrical stimulation, and so better resemble the intact brain than isolated (non-continuously stimulated) cultures. The proportion of GABAergic neurons in the cultures was significantly increased (p<1e-2, paired t-test) after burst-quieting for 2 days, suggesting that burst suppression operated through the homeostatic control of inhibitory neurotransmitter levels. We also studied the role of spontaneous bursts as potential carriers of information in the network by clustering these spatiotemporally diverse bursts. Spontaneous burst clusters were stable over hours and tetanic stimulation significantly reorganized the distribution of the clusters. In summary, this body of work explores the rules of network-level functional plasticity and provides the input (electrical stimulation) output (spatiotemporal patterns) mappings for behavioral studies in embodied hybrid systems. The results of this study may also have clinical implications in the development of sensory prostheses and treatment of diseases of aberrant network activity such as epilepsy.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-08-18T12:54:46Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Antonio Jorge Fontenele Neto.pdf: 9942923 bytes, checksum: 9de5bf466a9fc72acbc6a2a2d3a9c57c (MD5) / Made available in DSpace on 2016-08-18T12:54:46Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Antonio Jorge Fontenele Neto.pdf: 9942923 bytes, checksum: 9de5bf466a9fc72acbc6a2a2d3a9c57c (MD5) Previous issue date: 2015-08-25 / CAPEs / O córtex visual primário (V1) é a região do córtex cerebral responsável pela primeira etapa de processamento da informação visual capturada pela retina. Por ser uma das áreas corticais melhor compreendidas, V1 constitui um dos principais paradigmas de compreensão do processamento sensorial. Desde os anos 70 há uma extensa literatura que estuda propriedades de resposta de neurônios de V1, principalmente com eletrodos individuais e utilizando-se como modelo animal gatos e macacos. Tem-se conhecimento de onde partem seus principais inputs e quais estímulos fazem os neurônios dispararem (grades senoidais com determinadas frequências espaciais e temporais). Mais recentemente, com o uso de matrizes de eletrodos, se tornou possível a investigação de propriedades coletivas da atividade e codificação neurais, que não eram possíveis de serem desvendadas com eletrodos individuais. Além disso, no estado da arte tecnológico atual, o uso do rato como modelo animal permite o registro da atividade neural com os animais em comportamento livre (sem anestesia ou contenção). No entanto, pouco se sabe sobre especificidades das propriedades de resposta dos neurônios do córtex visual do rato. Este trabalho teve por objetivo desenvolver um aparato e um protocolo experimental no Laboratório de Neurociência de Sistemas e Computacional adequado para estudo das propriedades de resposta de neurônios de V1 de ratos usando matrizes de eletrodos. Finalmente, apresentamos resultados experimentais onde caracterizamos respostas de neurônios de V1 a diferentes estímulos visuais (Funções de Gabor ou Grades) seja em ruído denso ou rarefeito, variando as propriedades de frequências temporal e espacial de estimulação, densidades de estímulos, velocidade, etc. Concluímos que implementamos com sucesso a técnica experimental, que abre inúmeras perspectivas futuras de pesquisas nesta linha no Departamento de Física da Universidade Federal de Pernambuco. / The primary visual cortex (V1) is the cerebral cortex region responsible for the first processing step of the visual information captured by the retina. Being one of the most studied and well described cortical sensory areas, V1 is one of the main paradigms for the study of sensory processing. Since the 70s, there is a vast literature that studies properties of V1’s neurons, specially using single electrodes and using cats and monkeys as animal models. The anatomical conectivity of the visual pathway is known, from the retina to the lateral geniculate nucleus to V1, as well as the main visual stimulations that make V1 neurons fire (sinusoidal gratings with certain spatial and temporal frequencies). More recently, using multielectrode arrays, it became possible to study coletive properties of the activity and neural codification, that could not be unveiled with single electrodes. Furthermore with, the current state of the art in multielectrode recordings it is possible to record the neural activity in frelly behaving rats (without anesthesia or restraint). This represents an advantage in using the rat as animal model. However, little is known about specificities of the V1 neurons response properties in the rat. The aim of this work is to develop, in the Laboratório de Neurociência de Sistemas e Computacional, an apparatus and an experimental protocol suitable for the study of visual response properties of V1’s neurons in rats, using multielectrode array recordings. Finally, we present experimental results that characterize the response of V1’s neurons with different visual stimuli (Gabor or Grating Functions), either in dense os sparse noise modes, varying the spatial and temporal stimulation frequencies, stimulus density, speed, etc. We conclude that the experimental technique was implemented successfully. These results open important perspectives of future research on this field for the Departamento de Física at the Universidade Federal de Pernambuco.

No description available.