<p>In order to understand how the brain functions, under normal as well as pathological conditions, it is important to study the mechanisms underlying information integration. Depending on the nature of an input arriving at a synapse, different strategies may be used by the neuron to integrate and respond to the input. Naturally, if a short train of high-frequency synaptic input arrives, it may be beneficial for the neuron to be equipped with a fast mechanism that is highly sensitive to inputs on a short time scale. If, on the contrary, inputs arriving with low frequency are to be processed, it may be necessary for the neuron to possess slow mechanisms of integration. For example, in certain working memory tasks (e. g. delay-match-to-sample), sensory inputs may arrive separated by silent intervals in the range of seconds, and the subject should respond if the current input is identical to the preceeding input. It has been suggested that single neurons, due to intrinsic mechanisms outlasting the duration of input, may be able to perform such calculations. In this work, I have studied a mechanism thought to be particularly important in supporting the integration of low-frequency synaptic inputs. It is mediated by a cascade of events that starts with activation of group I metabotropic glutamate receptors (mGlu1/5), and ends with a membrane depolarization caused by a current that is mediated by canonical transient receptor potential (TRPC) ion channels. This current, denoted I_TRPC, is the focus of this thesis.</p><p>A specific objective of this thesis is to study the role of I_TRPC in the integration of synaptic inputs arriving at a low frequency, < 10 Hz. Our hypothesis is that, in contrast to the well-studied, rapidly decaying AMPA and NMDA currents, I_TRPC is well-suited for supporting temporal summation of such synaptic input. The reason for choosing this range of frequencies is that neurons often communicate with signals (spikes) around 8 Hz, as shown by single-unit recordings in behaving animals. This is true for several regions of the brain, including the entorhinal cortex (EC) which is known to play a key role in producing working memory function and enabling long-term memory formation in the hippocampus.</p><p>Although there is strong evidence suggesting that I_TRPC is important for neuronal communication, I have not encountered a systematic study of how this current contributes to synaptic integration. Since it is difficult to directly measure the electrical activity in dendritic branches using experimental techniques, I use computational modeling for this purpose. I implemented the components necessary for studying I_TRPC, including a detailed model of extrasynaptic glutamate concentration, mGlu1/5 dynamics and the TRPC channel itself. I tuned the model to replicate electrophysiological in vitro data from pyramidal neurons of the rodent EC, provided by our experimental collaborator. Since we were interested in the role of I_TRPC in temporal summation, a specific aim was to study how its decay time constant (τ_decay) is affected by synaptic stimulus parameters.</p><p>The hypothesis described above is supported by our simulation results, as we show that synaptic inputs arriving at frequencies as low as 3 - 4 Hz can be effectively summed. We also show that τ_decay increases with increasing stimulus duration and frequency, and that it is linearly dependent on the maximal glutamate concentration. Under some circumstances it was problematic to directly measure τ_decay, and we then used a pair-pulse paradigm to get an indirect estimate of τ_decay.</p><p>I am not aware of any computational model work taking into account the synaptically evoked I_TRPC current, prior to the current study, and believe that it is the first of its kind. We suggest that I_TRPC is important for slow synaptic integration, not only in the EC, but in several cortical and subcortical regions that contain mGlu1/5 and TRPC subunits, such as the prefrontal cortex. I will argue that this is further supported by studies using pharmacological blockers as well as studies on genetically modified animals.</p> / QC 20101005

Made available in DSpace on 2014-07-29T15:08:23Z (GMT). No. of bitstreams: 1 dissertacao fabio b rodrigues.pdf: 2707992 bytes, checksum: 8e5e7c8d4e6f975bb6e22b0b760695a6 (MD5) Previous issue date: 2010-04-23 / From motoneurons models (MN) of complex geometry and structure, able to reproduce the characteristics of a real motoneuron, the aim of this work is to verify the functional differences between proximal and distal synapses, investigating the nonlinearities in passive equivalent dendrites of motoneuron models and to study the influence of persistent calcium channels type L (Cav1.3), present in the dendrites, in the synaptic amplification. The original models developed by Vieira and Kohn (2007), implemented in C++, were expanded. This allowed to accomplish tests in order to verify the functional differences of synapses that occur near the soma and along of the dendrites. Persistent channels of calcium (CaV1.3) were modeled in dendrites and the influence of these channels in the amplification of the synaptic currents along the dendrites was verified. Finally, the nonlinearities of responses degree in the dendritic tree for different synaptic activation was evaluated. In order to verify the functional differences between proximal and distal synapses pure sinusoids were injected in different dendritic compartments of the models. The results showed attenuation at higher frequencies and the cutoff frequency is smaller as we move along the dendrites far from the soma. The influence of persistent calcium channels was verified comparing the functional differences between proximal and distal synapses along of the soma, with and without them. The initial results showed a notable amplification of synaptic activation. Nonlinear interactions were evaluated using sinusoids synaptic inputs with different frequencies in two or more equivalent dendrites in different compartments. The frequency spectrum of the current between the initial segment and the soma was analyzed by comparing the peak amplitude of harmonics and spurious bands with the peak amplitude of the fundamental frequency of smaller amplitude: the smaller these differences were, greater was degree of nonlinearity between synaptic activation in different dendritic segments. The results showed a high degree of nonlinearity between the dendrites. The cases where the synaptic conductance was varied showed greater degree of nonlinearity in relation to the cases in which the synaptic current was varied. / A partir de modelos de motoneurônios (MN) de geometria e estrutura complexa e capacidade de reproduzir as características de um motoneurônio real, este trabalho tem como objetivos verificar as diferenças funcionais entre sinapses proximais e distais, investigar as interações não lineares de ativações sinápticas e estudar a influência dos canais persistentes de Ca2+ tipo L existentes nos dendritos na amplificação sináptica. Para isso, os modelos originais desenvolvidos por Vieira e Kohn (2007), implementados em C++, foram expandidos. Isso possibilitou a realização de testes que verificaram as diferenças funcionais de sinapses que ocorrem em segmentos dendríticos distintos. Foram modelados canais de cálcio tipo L (CaV1.3) nos dendritos, verificando a influência destes na amplificação das correntes sinápticas e avaliando o grau de não linearidade da arborização dendrítica para diferentes ativações sinápticas. A verificação das diferenças funcionais entre as sinapses proximais e distais foi realizada, pela injeção de senoides puras em diferentes compartimentos dendríticos do modelo. Os resultados mostraram atenuações mais intensas nas altas frequências e frequência de corte mais baixa em compartimentos dendríticos mais distantes do soma. A influência dos canais persistentes de cálcio foi verificada comparando as diferenças funcionais entre sinapses proximais e distais ao soma com e sem os canais de cálcio persistentes. Os resultados apontam amplificação da ativação sináptica. As interações não lineares foram avaliadas aplicando potenciais senoidais pós-sinápticos com frequências primas entre si, em dois ou mais dendritos equivalentes simultaneamente e em compartimentos dendríticos diferentes. O espectro de frequência da corrente efetiva foi analisado, comparando a amplitude do pico das harmônicas e das raias espúrias com a amplitude do pico da frequência fundamental de menor amplitude: quanto menores estas diferenças maior o grau de não linearidade entre as ativações sinápticas em segmentos dendríticos distintos. Os resultados sugerem um alto grau de não linearidade entre os dendritos. Notou-se que, nas situações, quando variou-se a condutância sináptica, maior foi o grau de não linearidade em relação aos casos em que variou-se a corrente sináptica, sugerindo influência do potencial de membrana que introduz não linearidades na somação de correntes sinápticas.

In order to understand how the brain functions, under normal as well as pathological conditions, it is important to study the mechanisms underlying information integration. Depending on the nature of an input arriving at a synapse, different strategies may be used by the neuron to integrate and respond to the input. Naturally, if a short train of high-frequency synaptic input arrives, it may be beneficial for the neuron to be equipped with a fast mechanism that is highly sensitive to inputs on a short time scale. If, on the contrary, inputs arriving with low frequency are to be processed, it may be necessary for the neuron to possess slow mechanisms of integration. For example, in certain working memory tasks (e. g. delay-match-to-sample), sensory inputs may arrive separated by silent intervals in the range of seconds, and the subject should respond if the current input is identical to the preceeding input. It has been suggested that single neurons, due to intrinsic mechanisms outlasting the duration of input, may be able to perform such calculations. In this work, I have studied a mechanism thought to be particularly important in supporting the integration of low-frequency synaptic inputs. It is mediated by a cascade of events that starts with activation of group I metabotropic glutamate receptors (mGlu1/5), and ends with a membrane depolarization caused by a current that is mediated by canonical transient receptor potential (TRPC) ion channels. This current, denoted ITRPC, is the focus of this thesis. A specific objective of this thesis is to study the role of ITRPC in the integration of synaptic inputs arriving at a low frequency, &lt; 10 Hz. Our hypothesis is that, in contrast to the well-studied, rapidly decaying AMPA and NMDA currents, ITRPC is well-suited for supporting temporal summation of such synaptic input. The reason for choosing this range of frequencies is that neurons often communicate with signals (spikes) around 8 Hz, as shown by single-unit recordings in behaving animals. This is true for several regions of the brain, including the entorhinal cortex (EC) which is known to play a key role in producing working memory function and enabling long-term memory formation in the hippocampus. Although there is strong evidence suggesting that ITRPC is important for neuronal communication, I have not encountered a systematic study of how this current contributes to synaptic integration. Since it is difficult to directly measure the electrical activity in dendritic branches using experimental techniques, I use computational modeling for this purpose. I implemented the components necessary for studying ITRPC, including a detailed model of extrasynaptic glutamate concentration, mGlu1/5 dynamics and the TRPC channel itself. I tuned the model to replicate electrophysiological in vitro data from pyramidal neurons of the rodent EC, provided by our experimental collaborator. Since we were interested in the role of ITRPC in temporal summation, a specific aim was to study how its decay time constant (τdecay) is affected by synaptic stimulus parameters. The hypothesis described above is supported by our simulation results, as we show that synaptic inputs arriving at frequencies as low as 3 - 4 Hz can be effectively summed. We also show that τdecay increases with increasing stimulus duration and frequency, and that it is linearly dependent on the maximal glutamate concentration. Under some circumstances it was problematic to directly measure τdecay, and we then used a pair-pulse paradigm to get an indirect estimate of τdecay. I am not aware of any computational model work taking into account the synaptically evoked ITRPC current, prior to the current study, and believe that it is the first of its kind. We suggest that ITRPC is important for slow synaptic integration, not only in the EC, but in several cortical and subcortical regions that contain mGlu1/5 and TRPC subunits, such as the prefrontal cortex. I will argue that this is further supported by studies using pharmacological blockers as well as studies on genetically modified animals. / QC 20101005