Our sensory experiences are encoded in the patterns of activity of the neurons in our brain. While we know we are capable of sensing and responding to a constantly changing sensory environment, we often study neural activity by repeatedly presenting the same stimulus and analyzing the average neural response. It is not understood how the average neural response represents the dynamic neural activity that produces our perceptions. In this work, we use functional imaging of the rodent primary somatosensory cortex, specifically the whisker representations, and apply classic signal-detection methods to test the predictive power of the average neural response. Stimulus features such as intensity are thought to be perceptually separable from the average representation; however, we show that stimulus intensity cannot be reliably decoded from neural activity from only a single experience. Instead, stimulus intensity was encoded only across many experiences. We observed this probabilistic neural code in multiple classic sensory paradigms including complex temporal stimuli (pairs of whisker deflections) and multi-whisker stimuli. These data suggest a novel framework for the encoding of stimulus features in the presence of high-neural variability. Specifically we suggest that our brains can compensate for unreliability by encoding information redundantly across cortical space. This thesis predicts that a somatosensory stimulus is not encoded identically each time it is experienced; instead, our brains use multiple redundant pathways to create a reliable sensory percept.

Neocortical circuits can adapt to changes in sensory input by modifying the strength or number of synapses. These changes have been well-characterized electrophysiologically in primary somatosensory (barrel) cortex of rodents across different ages and with different patterns of whisker stimulation. Previous work from our lab has identified layer-specific critical periods for synaptic potentiation after selective whisker experience (SWE), where all but one row of facial whiskers has been removed. Although whole-cell patch-clamp recording methods enable a mechanistic understanding of how synaptic plasticity can occur in vivo, they are painstakingly slow, typically focus on a small number of observed events, and are focused on a single pathway or restricted anatomical area. For example, most studies of plasticity in barrel cortex have focused on analyses of experience-dependent synaptic changes in layer 4 and layer 2/3, at a single time point, but it is unclear whether such changes are limited to these layers, or whether they persist over long time periods. Here we employ an established electron-microscopic technique that selectively intensifies synaptic contacts, in combination with unbiased, automated synapse detection, to broadly explore experience-dependent changes in synaptic size and density across many neocortical layers, regions, and time periods in a high-throughput fashion. To validate the method, we focused on imaging synaptic contacts at time points surrounding the critical period for strengthening of excitatory synapses in mouse barrel cortex, and compared these to electrophysiological analyses that show a doubling of synaptic events targeting layer 2/3 pyramidal neurons following SWE. We found that the pattern of occurrence of synapses across the cortical layers is significantly different following SWE. Also, an increase in length was observed specifically in layer 3 synapses. Furthermore, we uncovered potential bidirectional plasticity in L6 synapses depending on the developmental state of circuit and a potential critical period onset for L5A synapse at PND 18. The high resolution imaging and unbiased synapse detection has enabled us to potentially tease apart synaptic changes that occur in a laminar specific fashion. This high-throughput method will facilitate analysis of experience-dependent changes in synaptic density by age, sensory experience, genotype, pharmacological treatments or behavioral training, and will enable classification of synaptic structure to identify key parameters that can be changed by these variables.

京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第13156号 / 論医博第2143号 / 新制||医||1028(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 林 康紀, 教授 渡邉 大, 教授 影山 龍一郎 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

Ratten verwenden Schnurrhaare (Vibrissen) zur Berührungswahrnehmung, und die Leitungsbahn von den Vibrissen zum primären somatosensorischen Areal (Barrel Cortex, BC) ist gut untersucht. Ratten zeigen auch vielfältiges Sozialverhalten, u.a. Berührung von Artgenossen mit ihren Vibrissen. Es ist jedoch unbekannt, wie diese sozialen Berührungssignale im Gehirn repräsentiert sind. Deshalb hatte die vorliegende Studie zum Ziel, die neuronale Repräsentation von sozialen Berührungen im BC zu untersuchen und mit anderer somatosensorischer Stimulation zu vergleichen. Mit extrazellulären Einzelzellableitungen in sich frei bewegenden Ratten habe ich gezeigt, dass die Aktivität eines Großteils von Neuronen im BC durch soziale Berührungen moduliert wird. Antworten waren meist erregend und Feuerraten während sozialer Interaktionen unterschieden sich zwischen kortikalen Schichten. Ratten bevorzugten Interaktionen mit Artgenossen gegenüber unbelebten Stimuli. Auch die Berührungsstrategien unterschieden sich, dabei wurden Objekte mit regelmäßigeren Bewegungen abgetastet, und die Vibrissen weiter vorgestreckt. Neuronale Antworten unterschieden sich ebenso, mit leicht aber konsistent schwächeren Antworten auf Objekte. Interessanterweise habe ich geschlechtsspezifische Unterschiede in neuronalen Antworten beobachtet. Der ausgeprägteste war die stärkere Modulation regulär-feuernder (RF) Zellen in Männchen während sozialer Berührungen. Dieser Unterschied konnte nicht mit sozialem Berührungsverhalten erklärt werden, was eventuell auf eine neurale Grundlage dieser Differenz hindeutet. Zudem feuerten RF-Zellen von Weibchen deutlich seltener, wenn das Weibchen im Östrus war. Zusammenfassend ist dies die erste Studie, die soziale Signale in einem primären sensorischen Areal bei sich frei bewegenden Tieren auf zellulärer Ebene untersuchte. Sie legt nahe, dass die Repräsentationen sensorischer Hirnrinde weniger stimulusabhängig und stärker top-down-moduliert sein könnten, als zuvor angenommen. / Rats use their stiff facial hairs (whiskers) for somatosensation, and the pathway from the whiskers to the primary somatosensory cortex (barrel cortex, BC) is well known. Rats also show diverse social behaviors, including touch of conspecifics with their whiskers. The representation of these social touch signals in the brain is however unknown. Thus, the present study aimed at characterizing the neuronal representation of social touch signals in BC and comparing them with non-social somatosensory stimulation. Using extracellular single-cell recordings in freely-moving rats, I could show that the activity of a large fraction of BC neurons is modulated by social touch. Responses were typically excitatory and the pattern of firing rates during interactions differed between cortical layers. Rats preferred interactions with alive conspecifics over inanimate stimuli. Whisking strategies also differed in that inanimate stimuli were whisked at with more regular movements from more protracted set angles. Neuronal responses were also different, such that objects elicited slightly but consistently weaker responses than alive rats. Interestingly, I observed sex-specific differences in neuronal responses. Prominently, there was stronger modulation by social touch in regular-spikers (RS) recorded from males. This could not be explained by behavioral measures, possibly indicating a neural origin of this difference. Further, RS from females fired much more weakly when females were in estrus. In summary, this is the first study that investigated social signals in a primary sensory area of freely-moving animals at the cellular level. It suggests that representations in sensory cortices might be less stimulus-driven and more top-down modulated than previously thought.

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Chez les rongeurs, le traitement par le cortex à tonneaux de l'information sensorielle en provenance des vibrisses est mal compris. En effet, malgré l'aide fournie par l'organisation de ce cortex en une reproduction stricte de la topographie de l'appareil sensoriel, il a été difficile jusqu'à présent d'identifier de façon indiscutable le système de filtrage linéaire et non-linéaire qu'utilisent les neurones du cortex à tonneaux durant leur traitement des scènes tactiles auxquelles ils sont exposés. Pour mieux identifier ces traitements corticaux, nous avons développé un système de stimulation vibrissale permettant d'appliquer des déflections sur un grand nombre de vibrisses indépendamment, dans toutes les directions possibles et ce à travers une vaste gamme fréquentielle. En utilisant ce dispositif de stimulation multivibrissale durant des enregistrements extracellulaires de l'activité électrique des neurones du cortex à tonneaux de rats anesthésiés, nous avons pu identifier plus précisément le filtrage linéaire des stimulations vibrissales, qui s'avère similaire pour tous les neurones que nous avons pu enregistrer. Par ailleurs, en explorant les aspects non-linéaires du traitement effectué par ces neurones, nous avons noté qu'ils se séparent en deux familles distinctes : d'un côté des neurones "locaux" qui se sont avérés sensibles à des contrastes locaux dans les déflections multivibrissales. De l'autre, des neurones "globaux" capables au contraire de détecter des situations où les déflections sont similaires pour de nombreuses vibrisses. Enfin, en effectuant d'autres enregistrements dans la couche II/III du cortex à tonneaux, cette fois à l'aide d'un microscope deux-photons, nous avons pu noter que les neurones appartenant aux familles locales et globales étaient séparés en groupes spatialement distincts et que la position spatiale des neurones était plus généralement étroitement liée à l'ensemble de leurs propriétés de filtrage des déflections vibrissales.

It is well established that, following adaptation, cells adjust their sensitivity to reflect the global stimulus conditions. Two recent studies in guinea pig inferior colliculus (IC, Dean, Harper & McAlpine 2005) and rat barrel cortex (Garcia-Lazaro, Ho, Nair & Schnupp 2007) found that neural stimulus-response functions were displaced laterally in a manner that was dependent on the mean adapting stimulus. However, the direction of gain change, following adaptation to variance, was in contradiction to Information Theory, which predicts a decrease in gain with increased stimulus variance. On further analysis of the experimental data, presented within this thesis, it was revealed that the adaptive gain changes to global stimulus variance were, in fact, in the direction predicted by Information Theory. However, following adaptation to global mean amplitude, neural threshold was displaced to centre the SRF on inputs that were located on the edge of the stimulus distribution. It was found that adaptation scaled neural output such that the relationship between firing rate and local, as opposed to global, differences in stimulus amplitude was maintained; with the majority of cells responding to large differences in stimulus amplitude, on the 40ms scale. A small majority of cells responded to step-size differences, in amplitude, of either direction and were classed as novelty preferring. Adaptation to global mean was replicated in model neuron with spike-rate adaptation and tonic inhibition, which increased with stimulus mean. Adaptation to stimulus variance was replicated in three models 1: By increasing, in proportion to stimulus variance, background, excitatory and inhibitory firing rates in a balanced manner (Chance, Abbott & Reyes 2002), 2: A model of asymmetric synaptic depression (Chelaru & Dragoi 2008) and 3: a model combining non-linear input with synaptic depression. The results presented, within this thesis, demonstrate that neurons change their coding strategies depending upon the global levels of mean and variance within the sensory input. Under low noise conditions, neurons act as deviation detectors, i.e. are primed to respond to large changes in the stimulus on the tens of millisecond; however, under conditions of increased noise switch their encoding strategy in order to compute the full range of the stimulus distribution through adjusting neural gain.

Sensory signals are represented and propagated as spiking activity in multiple neuronal populations to lead to cognitive or motor behavior in organisms. Neural processing underlying sensory-motor behavior is understood by uncovering the governing computational principles and the biophysical mechanisms that implement the principles. While these mechanisms have been studied extensively at the single-neuron and system levels, activity within neuronal networks significantly impact neural processing. For example, there are spatiotemporal interactions (neural correlations) between responses of neurons within populations that could potentially impact signal representation and propagation. Furthermore, the effects of associative plasticity are also expected to alter network activity and its propagation. The effects of plasticity on network activity cannot be predicted from individual neuronal responses due to the complex, non-linear interactions within neuronal networks. Thus examining neural correlations in network activity and the propagation of network activity, requires recording spiking activity from large, heterogeneous, populations of spatially distributed neurons simultaneously. Studies addressing the propagation of network activity have been limited to theoretical approaches. Empirical studies have been limited by the technical difficulties in recording from a large number of neurons simultaneously. To overcome this challenge we developed a novel technique, dithered random-access functional calcium imaging. This imaging technique records and extracts suprathreshold activity from a large number of neurons. This technique also has a high spike detection efficiency and millisecond temporal precision. We applied this technique to measure the propagation of activity and neural correlations in activity evoked by afferent, thalamocortical inputs in the recurrent cortical networks of the mouse barrel cortex. We found that the cortical activity evoked by novel (naïve), thalamocortical inputs showed limited propagation of activity and decrease in propagation of neural correlations (measured from neuronal pairs within each population) from L4 to L2/3 network of the responding column. However, associative cortical plasticity was induced from pairing thalamocortical inputs with intracortical inputs. This pairing resulted in increased propagation of activity. The pairing also modified the propagation of neural correlations. Our results suggest that synaptic plasticity in intracortical circuits contributes to the modified propagation of activity and neural correlations. The modified propagation of neural correlations could in turn contribute to behavioral performance in vivo following perceptual learning. / text

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / Em roedores, as vibrissas são detectores táteis que desempenham papel importante na exploração espacial do ambiente e na discriminação de texturas. No córtex somatosensorial, os campos receptivos de cada uma das vibrissas estão organizados no hemisfério contralateral em colunas discretas denominadas barris. A lesão unilateral dos barris produz um comportamento assimétrico caracterizado pela redução no uso da vibrissa contralateral à lesão na exploração do ambiente, assimetria esta que diminui progressivamente na medida em que os animais são repetidamente testados. Em ratos, este comportamento, normalmente medido pelo número de vezes que os animais encostam as vibrissas na parede de um campo aberto, tem se mostrado uma ferramenta importante em estudos de plasticidade e recuperação funcional após lesões corticais. Contudo, em camundongos com lesões unilaterais dos barris, o registro dos toques das vibrissas na parede tem levado a resultados contraditórios. Esse trabalho tem por objetivo principal o estabelecimento de um modelo comportamental para avaliação da recuperação funcional após lesões unilaterais dos barris do córtex somatosensorial em camundongos. Para tanto, o sentido dos deslocamentos realizados próximos às quinas do campo aberto foi registrado em camundongos Suíços machos submetidos à criolesão unilateral dos barris foi avaliado em três estudos independentes. No primeiro estudo, demonstramos que no grupo Criolesado houve um predomínio dos deslocamentos em sentido contralateral na primeira vez em que foram testados no campo aberto e este resultado foi independente do fato de na primeira sessão ter sido realizada um ou nove dias após a cirurgia. Além disso, demonstramos que o predomínio de deslocamentos em sentido contralateral foi diminuindo na medida em que os animais eram repetidamente testados no campo aberto. No segundo estudo, demonstramos que os animais do grupo Criolesado que foram previamente submetidos a cinco sessões experimentais no campo aberto não apresentaram, após a cirurgia, diferenças entre os deslocamentos realizados em sentido ipsolateral e contralateral à lesão. Já no terceiro estudo, demonstramos que os animais do grupo Criolesado que não foram previamente testados no campo aberto apresentam um predomínio de deslocamentos em sentido contralateral, mesmo quando o teste foi realizado 48 dias após a lesão unilateral dos barris. Nossos dados sugerem que o sentido dos deslocamentos próximo às quinas do campo aberto pode ser uma ferramenta importante para avaliar a recuperação das lesões unilaterais nos barris do córtex somatosensorial. Além disso, para avaliar a recuperação funcional após a lesão unilateral dos barris do córtex somatossensorial, sem o viés da habituação à situação do teste, os animais devem ser testados apenas uma vez / In rodents, the vibrissae are tactile detectors that have an important role in the spatial exploration of the environment and in texture discrimination. In the somatosensory cortex, the receptive fields of each of the vibrissae are organized into discrete columns, known as barrels, in the contralateral hemisphere. The unilateral lesions of the barrels results in an asymmetric behavior characterized by the reduction in the use of the vibrissae contralateral to the lesion during the exploration of the environment. This asymmetry progressively diminishes as the animal is repeatedly tested in the same environment. In rats, this behavior, usually assessed by the number of times the animals touch the walls of an open field arena with their vibrissae, has been considered a useful tool in studies addressing plasticity and functional recovery after cortical lesions. However, in mice with unilateral lesions of the barrels, the analysis of the number of times that the vibrissae touch the walls have lead to contradictory results. The present work aims to establish a behavioral model for the evaluation of functional recovery after unilateral lesions of the barrel field located in the somatosensory cortex of the mouse. To that end, the direction of movement near the corners of the open field of Swiss male mice subjected to unilateral cryolesion of the barrel was analyzed in three independent studies. In the first study, we demonstrated that the cryolesioned animals displayed a predominance of contralateral movements in the first time that they were tested in the open field and that this result was present whether the animals was first tested one or nine days after the lesion. Furthermore, we demonstrated that the predominance of movements toward the contralateral side of the lesion diminished as the animals were repeatedly tested in the open field. In the second study, we demonstrated that the cryolesioned animals that were, prior to barrel field lesioning, subjected to five sessions in the open field did not display, after lesioning, differences between the number of ipso and contralateral movements. In the third study, we demonstrated that the cryolesioned animals that were not previously tested in the open field displayed a predominance of contralateral movements even if the first test was carried out forty eight days after the unilateral lesion of the barrels. Our data suggest that the analysis of the direction of movement near the corners of the open field can be an important tool in the assessment of functional recovery after unilateral lesions of the barrels located in the somatosensory cortex of mice. Moreover, in order to properly assess functional recovery after unilateral lesion of the barrel field without the confounding factor of habituation, animals must be tested only once