Mechanical loading can modulate tissue plasticity and has potential applications in rehabilitation science and regenerative medicine. To safely and effectively introduce mechanical loads to human cells, tissues, and the entire body, we need to understand the optimal loading environment to promote growth and health. The purpose of this research was 1) to validate a limb vibration and compression system; 2) to determine the effect of limb vibration on neural excitability measured by sub-threshold TMS-conditioned H-reflexes and supra-threshold TMS; 3) to determine changes in center of pressure, muscle activity, and kinematics during a postural task following limb vibration; 4) to determine the effect of vibration on accuracy and long latency responses during a weight bearing visuomotor task. The major findings of this research are 1) the mechanical system presented in the manuscript can deliver limb vibration and compression reliably, accurate, and safely to human tissue; 2) sub-threshold cortical stimulation reduces the vibration-induced presynaptic inhibition of the H-reflex. This reduction cannot be attributed to an increase in cortical excitability during limb vibration because the MEP remains unchanged with limb vibration; 3) limb vibration altered the soleus and tibialis EMG activity during a postural control task. The vibration-induced increase in muscle activity was associated with unchanged center of pressure variability and reduced center of pressure complexity; 4) healthy individuals were able to accommodate extraneous afferent information due to the vibration interventions They maintained similar levels of accuracy of a visuomotor tracking task and unchanged long latency responses during an unexpected perturbation.

In order for stroke subjects to gain functional recovery of their hemiparetic limbs, facilitation techniques such as the repetitive facilitation exercise, or RFE, have been developed. Currently, there is a lack of understanding of the neural mechanisms associated with these types of facilitation techniques. To better understand the neural mechanisms associated with the RFE a functional magnetic resonance imaging (fMRI) study should be conducted. This thesis presents experimental results testing the feasibility of implementing an fMRI-compatible actuator to facilitate a myotatic reflex in synchronization with the subject’s intention to move their hemiparetic limb. Preliminary data from a healthy individual demonstrated the feasibility of overlapping the long latency component of the afferent myotatic reflex, created by electrical stimulation, with descending nerve impulses, created using transcranial magnetic stimulation, in a time window of 15ms. In addition, a pneumatic actuation time delay due to long transmission line was evaluated. The pneumatic actuator met the timing precision requirement for the rehabilitation device for varying transmission line lengths. Therefore a pneumatic actuation system was chosen for the rehabilitation device. This thesis will also presents on the design of an fMRI-compatible pneumatic actuator device to excite a stretch reflex response. Initial, experimental results with the device demonstrated that the designed pneumatic device can control the timing of the muscle response with a fixed signal within the required 15ms window required for cortical facilitation, which was found in the previous feasibility study. However, the device was unable to create a long latency reflex observable at the muscle. Finally, this thesis presents on the capability of the device in creating subthreshold long latency response with precision to overlap with a subthreshold descending nerve impulse, created using transcranial magnetic stimulation. The overlap of the two responses was evaluated by comparing the amplitude of the muscle response with and without the stretch reflex, created by the fMRI-compatible pneumatic actuator device. Varying time delays were analyzed.

A broad understanding of motor control has been achieved through research performed on upper extremity reaching, walking on level ground, and static balance. Though invaluable insights have been achieved under these testing paradigms, inherent limitations result in less being known regarding functional movement in weight-bearing. Gait studies require large numbers of consecutive steps to achieve high reliability, static balance is limited to the goal of no movement, and upper extremity reaching lacks insights into feedback from the vestibular system. Here we describe (and provide a supplemental video of) a system for testing and training the performance of a weight-bearing, visuomotor task in the form of a mini-squat according to a sinusoidal trace on a screen. In this work, we determined that by altering both task movement rate and resistance at the knee, a hierarchy of difficulty was achieved at all ages. As age increases, there is a velocity-error tradeoff; speed of movement is attempted to be maintained while error is sacrifieced. When introducing an unexpected force perturbation (rapid release of the resistance of the squat for less than a second), older adults who are least able to match the frequency of the task experience the greatest error and velocity rates during the perturbation. This exposes a possible deficit in the feedback control system of even healthy older adults, where future studies may determine if early intervention to prevent such changes may prevent future injury and disability. When older and younger adults learned to perform the visuomotor task while performing a simultaneous cognitive task, learning was slowed as complexity of the cognitive task increased. In older adults, a difficult cognitive task inhibited acquisition of the squatting task with no apparent improvement in trial error nor coherence. Upon retesting of the motor task, there was no difference between dual-task and single-task trained ability to consolidate the motor task in both age groups, though those that trained under a dual-task condition were more able to automate the motor task as measured by a smaller dual-task cost. This may indicate that dual-task training leads to freeing cognitive resources from attending to a functional movement so that they may attend to other tasks such as what may be happening in the environment. Finally, executive function as measured by the Flanker Test, explained 80% of the variability of final day visuomotor error, being a possible prognostic factor for dual-task interventions. Future directions will determine if increased automaticity of a mini-squat will lead improvement to overall improved functional mobility and reduced lower extremity injuries when functioning in a busy community.

Introdução: O implante coclear proporciona aos indivíduos com perda auditiva severa e severa a profunda, que não se beneficiam do uso de aparelho de amplificação sonora individual, maior habilidade quanto à comunicação. Os potenciais evocados auditivos de longa latência (N1, P2, N2 e P300) podem ser um indicador diferencial na avaliação dos usuários de implante coclear. Esses potenciais fornecem parâmetros objetivos de geradores neurais, relacionados às habilidades de atenção, discriminação e memorização de estímulos tonais ou de fala. No estudo, os potenciais N1, P2, N2 e P300, gerados por estímulos de fala em usuários de implante coclear, foram caracterizados e posteriormente correlacionados às variáveis: percepção de fala, tempo de surdez e tempo de uso do implante. Metodologia: A pesquisa foi desenvolvida na Clínica de Fonoaudiologia da Faculdade de Odontologia de Bauru e no Centro de Pesquisas Audiológicas do Hospital de reabilitação de Anomalias, ambos unidades da Universidade de São Paulo. Os potenciais foram captados por eletrodos posicionados nas regiões Cz/Fz/M1/M2 da calvária e conectados ao Biologic\'s Evoked Potential System (EP) por cabos, seguindo os padrões do sistema internacional 10-20. Participaram da pesquisa 25 usuários de implante coclear, portadores de surdez pré-lingual, na faixa etária de 6,6 a 12,3 anos. Para realização do procedimento os indivíduos foram orientados a levantar a mão assim que ouvissem o estímulo raro (/a/ e /da/), apresentado na proporção de 20/80 em relação ao estímulo freqüente (/i/ e /ba/). Os estímulos de fala /ba-da/ e /i-a/ foram apresentados em oddball paradigma, a uma intensidade de 60 dB NPS por uma caixa acústica. Resultados: Foram observadas correlações estatisticamente significantes entre a latência do potencial N1 e o tempo de surdez e de uso do implante; entre amplitude do potencial P2 e o tempo de surdez e tempo de uso do implante; entre a latência do potencial P300 e o reconhecimento de palavras. A latência média do N1 aumentou 6 ms por ano em relação ao tempo de surdez, mantido o tempo de uso do implante constante e diminuiu 3 ms de acordo com o aumento de 1 ano no tempo de uso. A amplitude média do P2 diminuiu 1 ?V, com o aumento do tempo de surdez em 1 ano, mantendo-se o tempo de uso fixo e aumentou, 0,5 ?V com o prolongamento de 1 ano no tempo de uso do IC, mantido o tempo de surdez constante. A latência média do potencial P300 aumentou 2 ms, de acordo com a piora do escore de reconhecimento de palavras,e diminuiu 2 ms proporcionalmente à melhora do mesmo escore. Conclusões: As variáveis: tempo de surdez e de uso do implante, influenciaram os valores de latência e amplitude dos potenciais N1 e P2 , sugerindo plasticidade cortical. A latência do potencial P300 refletiu os processos cognitivos relacionados às habilidades de atenção e discriminação auditiva e apresentou relação de significância com os escores de percepção de fala nos usuários de implante coclear / Introduction: Cochlear implants supply subjects presented with severe and profound hearing loss, who do not benefit from the hearing aid, with a greater communication ability. The long latency auditory evoked potentials (N1, P2, N2 and P300) are a differential indicator in the assessment of cochlear implant users. These potentials provide objective parameters of neural generators related to the skills of attention, discrimination and memorization of tonal or speech stimuli. In this study, potentials N1, P2, N2 and P300, generated through speech stimuli in cochlear implant users, were characterized and correlated with the variables speech perception, time of deafness and time of cochlear implant use. Methodology: the research was carried out at the Speech Pathology clinic of the Dental School and at the Audiological Research Center with the Craniofacial Anomalies Rehabilitation Hospital, both with University of São Paulo at Bauru, SP. The potentials were recorded by electrodes placed in regions Cz/Fz/M1/M2 of the skull and connected to the Biologic\'s Evoked Potential System (EP) through cables, according to the standards of international system 10-20. Twenty-five cochlear implant users, presented with pre-lingual deafness, in the age range 6.6 - 12.3 yrs, participated in the study. The subjects were requested to raise their hand as soon as they heard the rare stimulus (/a/ and /da/), presented in the proportion 20/80 in relation to the frequent stimulus (/i/ and /ba/). The speech stimuli /ba-da/ and /i-a/ were presented in oddball paradigm, at an intensity of 60 dB NPS, through a speaker. Results: statistically significant correlations were observed between the latency of potential N1 and the time of deafness and time of cochlear implant use; between amplitude of potential P2 and the time of deafness and time of cochlear implant; between the latency of potential P300 and word recognition. The mean latency of N1 increased 6 ms per year in relation to the time of deafness, with the time of implant use constant and diminished 3 ms according to the increase of 1 year in the time of use. The mean amplitude of P2 diminished 1 ?V, as the time of deafness increased in a year, by keeping fixed the time of use, and increased 0.5 ?V, with the increase of a year in the time of CI use, by keeping constant the time of deafness. The mean latency of potential P300 increased 2 ms as the word recognition score worsened and diminished 2 ms proportionally to the improvement of the same score. Conclusions: The variables time of deafness and cochlear implant use influenced the latency and amplitude of potentials N1 and P2, suggesting a cortical plasticity. The latency of potential P300 reflected the cognitive processes related to skills of auditory attention and discrimination, and presented a significance relation with the word perception score, in the subjects evaluated

OBJETIVO: verificar a inter-relação entre a recuperação postural e o bom ou baixo desempenho de indivíduos usuários de implante coclear (IC). MÉTODO: Foram avaliados 21 indivíduos divididos em três grupos: nove usuários de IC com bom desempenho, cinco de IC com baixo desempenho e sete controles. Todos os sujeitos foram submetidos a dois testes: Teste de Integração Sensorial (TIS) da Posturografia Dinâmica Computadorizada (PDC), com intervalo de 40 a 60 minutos entre eles, com a finalidade de observar o aprendizado de estratégias posturais no segundo teste. Os resultados obtidos foram submetidos à avaliação estatística. RESULTADOS: Os indivíduos com baixo desempenho apresentaram níveis significativamente mais baixos de aprendizado que os outros dois Grupos em C4, C5, C6 e IE. CONCLUSÂO: Os usuários de IC com bom desempenho mostraram melhor índice de recuperação postural quando comparados àqueles de IC com baixo desempenho / OBJECTIVE: To test the inter-relationship between postural recovery and good or poor performance of individuals using cochlear implant (CI). METHODS: We studied 21 subjects divided into three groups: nine cochlear implant users with good performance, 5 CI users with poor performance and 7 controls. All subjects underwent two tests in Sensory Integration Test (SOT) of computerized dynamic posturography (CDP) with an interval of 40 to 60 minutes between them, in order to observe the learning of postural strategies in the second test. The results were submitted to statistical evaluation. RESULTS: Individuals with poor performance had significantly lower difference between the results of the second and the first test than the other two groups at C4, C5, C6 and composite. CONCLUSION: CI users with good performance had better postural recovery rate when compared to the cochlear implant users with poor performance

Introdução: O implante coclear proporciona aos indivíduos com perda auditiva severa e severa a profunda, que não se beneficiam do uso de aparelho de amplificação sonora individual, maior habilidade quanto à comunicação. Os potenciais evocados auditivos de longa latência (N1, P2, N2 e P300) podem ser um indicador diferencial na avaliação dos usuários de implante coclear. Esses potenciais fornecem parâmetros objetivos de geradores neurais, relacionados às habilidades de atenção, discriminação e memorização de estímulos tonais ou de fala. No estudo, os potenciais N1, P2, N2 e P300, gerados por estímulos de fala em usuários de implante coclear, foram caracterizados e posteriormente correlacionados às variáveis: percepção de fala, tempo de surdez e tempo de uso do implante. Metodologia: A pesquisa foi desenvolvida na Clínica de Fonoaudiologia da Faculdade de Odontologia de Bauru e no Centro de Pesquisas Audiológicas do Hospital de reabilitação de Anomalias, ambos unidades da Universidade de São Paulo. Os potenciais foram captados por eletrodos posicionados nas regiões Cz/Fz/M1/M2 da calvária e conectados ao Biologic\'s Evoked Potential System (EP) por cabos, seguindo os padrões do sistema internacional 10-20. Participaram da pesquisa 25 usuários de implante coclear, portadores de surdez pré-lingual, na faixa etária de 6,6 a 12,3 anos. Para realização do procedimento os indivíduos foram orientados a levantar a mão assim que ouvissem o estímulo raro (/a/ e /da/), apresentado na proporção de 20/80 em relação ao estímulo freqüente (/i/ e /ba/). Os estímulos de fala /ba-da/ e /i-a/ foram apresentados em oddball paradigma, a uma intensidade de 60 dB NPS por uma caixa acústica. Resultados: Foram observadas correlações estatisticamente significantes entre a latência do potencial N1 e o tempo de surdez e de uso do implante; entre amplitude do potencial P2 e o tempo de surdez e tempo de uso do implante; entre a latência do potencial P300 e o reconhecimento de palavras. A latência média do N1 aumentou 6 ms por ano em relação ao tempo de surdez, mantido o tempo de uso do implante constante e diminuiu 3 ms de acordo com o aumento de 1 ano no tempo de uso. A amplitude média do P2 diminuiu 1 ?V, com o aumento do tempo de surdez em 1 ano, mantendo-se o tempo de uso fixo e aumentou, 0,5 ?V com o prolongamento de 1 ano no tempo de uso do IC, mantido o tempo de surdez constante. A latência média do potencial P300 aumentou 2 ms, de acordo com a piora do escore de reconhecimento de palavras,e diminuiu 2 ms proporcionalmente à melhora do mesmo escore. Conclusões: As variáveis: tempo de surdez e de uso do implante, influenciaram os valores de latência e amplitude dos potenciais N1 e P2 , sugerindo plasticidade cortical. A latência do potencial P300 refletiu os processos cognitivos relacionados às habilidades de atenção e discriminação auditiva e apresentou relação de significância com os escores de percepção de fala nos usuários de implante coclear / Introduction: Cochlear implants supply subjects presented with severe and profound hearing loss, who do not benefit from the hearing aid, with a greater communication ability. The long latency auditory evoked potentials (N1, P2, N2 and P300) are a differential indicator in the assessment of cochlear implant users. These potentials provide objective parameters of neural generators related to the skills of attention, discrimination and memorization of tonal or speech stimuli. In this study, potentials N1, P2, N2 and P300, generated through speech stimuli in cochlear implant users, were characterized and correlated with the variables speech perception, time of deafness and time of cochlear implant use. Methodology: the research was carried out at the Speech Pathology clinic of the Dental School and at the Audiological Research Center with the Craniofacial Anomalies Rehabilitation Hospital, both with University of São Paulo at Bauru, SP. The potentials were recorded by electrodes placed in regions Cz/Fz/M1/M2 of the skull and connected to the Biologic\'s Evoked Potential System (EP) through cables, according to the standards of international system 10-20. Twenty-five cochlear implant users, presented with pre-lingual deafness, in the age range 6.6 - 12.3 yrs, participated in the study. The subjects were requested to raise their hand as soon as they heard the rare stimulus (/a/ and /da/), presented in the proportion 20/80 in relation to the frequent stimulus (/i/ and /ba/). The speech stimuli /ba-da/ and /i-a/ were presented in oddball paradigm, at an intensity of 60 dB NPS, through a speaker. Results: statistically significant correlations were observed between the latency of potential N1 and the time of deafness and time of cochlear implant use; between amplitude of potential P2 and the time of deafness and time of cochlear implant; between the latency of potential P300 and word recognition. The mean latency of N1 increased 6 ms per year in relation to the time of deafness, with the time of implant use constant and diminished 3 ms according to the increase of 1 year in the time of use. The mean amplitude of P2 diminished 1 ?V, as the time of deafness increased in a year, by keeping fixed the time of use, and increased 0.5 ?V, with the increase of a year in the time of CI use, by keeping constant the time of deafness. The mean latency of potential P300 increased 2 ms as the word recognition score worsened and diminished 2 ms proportionally to the improvement of the same score. Conclusions: The variables time of deafness and cochlear implant use influenced the latency and amplitude of potentials N1 and P2, suggesting a cortical plasticity. The latency of potential P300 reflected the cognitive processes related to skills of auditory attention and discrimination, and presented a significance relation with the word perception score, in the subjects evaluated

OBJETIVO: verificar a inter-relação entre a recuperação postural e o bom ou baixo desempenho de indivíduos usuários de implante coclear (IC). MÉTODO: Foram avaliados 21 indivíduos divididos em três grupos: nove usuários de IC com bom desempenho, cinco de IC com baixo desempenho e sete controles. Todos os sujeitos foram submetidos a dois testes: Teste de Integração Sensorial (TIS) da Posturografia Dinâmica Computadorizada (PDC), com intervalo de 40 a 60 minutos entre eles, com a finalidade de observar o aprendizado de estratégias posturais no segundo teste. Os resultados obtidos foram submetidos à avaliação estatística. RESULTADOS: Os indivíduos com baixo desempenho apresentaram níveis significativamente mais baixos de aprendizado que os outros dois Grupos em C4, C5, C6 e IE. CONCLUSÂO: Os usuários de IC com bom desempenho mostraram melhor índice de recuperação postural quando comparados àqueles de IC com baixo desempenho / OBJECTIVE: To test the inter-relationship between postural recovery and good or poor performance of individuals using cochlear implant (CI). METHODS: We studied 21 subjects divided into three groups: nine cochlear implant users with good performance, 5 CI users with poor performance and 7 controls. All subjects underwent two tests in Sensory Integration Test (SOT) of computerized dynamic posturography (CDP) with an interval of 40 to 60 minutes between them, in order to observe the learning of postural strategies in the second test. The results were submitted to statistical evaluation. RESULTS: Individuals with poor performance had significantly lower difference between the results of the second and the first test than the other two groups at C4, C5, C6 and composite. CONCLUSION: CI users with good performance had better postural recovery rate when compared to the cochlear implant users with poor performance

Les actions thérapeutiques des antidépresseurs, disponibles actuellement, requièrent plusieurs semaines de traitement. Ce délai est dû aux adaptations des sites pré et post-synaptiques qui, respectivement, augmentent la disponibilité synaptique des monoamines sérotonine et noradrénaline (5-HT et NA), et entraînent les changements neuroplastiques modifiant la fonction neuronales dans les régions limbiques. Il a été récemment observé, chez un modèle animal de dépression, que l’agoniste RS67333 des récepteurs sérotoninergiques de type 5-HT4 produisait des changements comportementaux, électrophysiologiques, cellulaires et biochimiques, tel qu’observé chez les antidépresseurs. Ces changements apparaissent seulement après 3 jours de traitement tandis que les antidépresseurs nécessitent souvent plusieurs semaines. De plus, l’activation des récepteurs 5-HT4 ne générait pas de tolérance, et cela pendant 21 jours de traitement. Seulement, les propriétés de signalisation et de régulation de ces récepteurs sont très loin d’êtres établies. Nous avons alors voulu mieux caractériser ces deux aspects de leur fonction, en se concentrant d’avantage sur les isoformes a et b, fortement exprimés dans le système limbique. Pour cela, nous avons voulu évaluer d’abord leur capacité de production d’AMPc dans un système hétérologue. Les essais d’accumulation d’AMPc démontrent que les deux isoformes sont capables de moduler positivement et négativement des niveaux d’AMPc en présence de 5-HT. Par contre, la stimulation au RS67333 induit seulement une augmentation du niveau d’AMPc dans les deux cas. Ensemble, ces observations indiquent que les deux isoformes sont capables de coupler à l’adénylate cyclase à travers les protéines Gαs et Gαi. La quantification des récepteurs internalisés a montré que l’isoforme b internalisait plus efficacement que l’isoforme a suite à l’incubation à la 5-HT (61 ± 3 % pour le b vs 40 ± 2 % pour le a). Les protéines kinases PKA et PKC n’étaient pas impliquées dans cette différence, toutefois, la PKC a été trouvée essentielle à l’internalisation des deux isoformes. L’internalisation de l’isoforme b par 5-HT n’a pas été affecté par la surexpression de forme inactive de GRK2 (GRK2- K220R) et a été partiellement inhibé par un mutant négative de la β-arrestine (βarr(319-418)), tandis que l’internalisation de l’isoforme a a été bloquée par les deux. Ces observations indiquent que les mécanismes d’internalisation des deux isoformes du récepteur 5-HT4 les plus abondants dans le système nerveux central sont distincts. Des comportements spécifiques à chaque isoforme ont aussi été constatés au niveau de la régulation fonctionnelle suite à l’exposition au RS67333, qui désensibilise seulement l’isoforme b. D’après nos observations, nous avons conclu que les isoformes a et b diffèrent dans leur propriétés de signalisation et de régulation. L’incapacité du RS67333 à désensibiliser l’isoforme a fournit un substrat moléculaire pour les effets antidépressifs prolongés de cet agoniste dans les études pré-cliniques. / The therapeutic actions of antidepressants, currently available, require several weeks of treatment. This delay is due to pre-and post-synaptic sites adjustments, which, respectively, increase the availability of synaptic monoamines 5-HT and NA, and induce neuroplastic changes amending neuronal function in the limbic regions. We have recently observed in animal model of depression that serotonergic 4 receptor agonist RS67333 produces behavioural, electrophysiological, cellular and biochemical changes, as seen in antidepressants. More importantly, these changes appear only after 3 days of treatment while antidepressants often require several weeks. Moreover, activation of 5-HT4 receptors does not generate tolerance and that for 21 days of treatment. However, the signalling and regulation properties of these receptors haven’t been established yet. Here, we wanted to better characterize these two aspects of their function, and in particular for isoforms a and b, strongly expressed in the limbic system. First, we assessed their ability to produce cAMP in a heterologous system. Functional assays revealed that both isoforms were capable of positive and negative modulation of cAMP levels by 5-HT. Stimulation by RS67333 induced cAMP stimulation. Together, these observations indicate that that both isoforms are able to couple adenylate cyclase through Gαs and Gαi proteins. Moreover, quantification of receptors sequestration showed that isoform b internalised more efficiently than the isoform a, following incubation with 5-HT (61 ± 3 % for b, 40 ± 2 % for a). PKA and PKC proteins, two seconds messenger kinases activated by these receptors, are not involved in producing this difference. However, PKC is essential to the internalization of both isoforms. Isoform b internalization was not affected by an inactive GRK2 mutant (GRK2-K220R) and was partially inhibited by dominant negative β-arrestin (βarr(319-418), while isoform a internalization was dependant on both. These observations indicate that internalization mechanisms of the two isoforms, most abundantly expressed in the central nervous system, are different. Functional desensitization studies showned that RS67333 selectively desensitizes the isoform b, but not isoforme a. Based on these observations, we conclude that isoform a and b differ in their signalling and regulatory properties. Isoform a incapacity to desensitization by RS67333 provides a molecular substrate for prolonged antidepressant affects of this agonist in pre-clinical studies.

Les actions thérapeutiques des antidépresseurs, disponibles actuellement, requièrent plusieurs semaines de traitement. Ce délai est dû aux adaptations des sites pré et post-synaptiques qui, respectivement, augmentent la disponibilité synaptique des monoamines sérotonine et noradrénaline (5-HT et NA), et entraînent les changements neuroplastiques modifiant la fonction neuronales dans les régions limbiques. Il a été récemment observé, chez un modèle animal de dépression, que l’agoniste RS67333 des récepteurs sérotoninergiques de type 5-HT4 produisait des changements comportementaux, électrophysiologiques, cellulaires et biochimiques, tel qu’observé chez les antidépresseurs. Ces changements apparaissent seulement après 3 jours de traitement tandis que les antidépresseurs nécessitent souvent plusieurs semaines. De plus, l’activation des récepteurs 5-HT4 ne générait pas de tolérance, et cela pendant 21 jours de traitement. Seulement, les propriétés de signalisation et de régulation de ces récepteurs sont très loin d’êtres établies. Nous avons alors voulu mieux caractériser ces deux aspects de leur fonction, en se concentrant d’avantage sur les isoformes a et b, fortement exprimés dans le système limbique. Pour cela, nous avons voulu évaluer d’abord leur capacité de production d’AMPc dans un système hétérologue. Les essais d’accumulation d’AMPc démontrent que les deux isoformes sont capables de moduler positivement et négativement des niveaux d’AMPc en présence de 5-HT. Par contre, la stimulation au RS67333 induit seulement une augmentation du niveau d’AMPc dans les deux cas. Ensemble, ces observations indiquent que les deux isoformes sont capables de coupler à l’adénylate cyclase à travers les protéines Gαs et Gαi. La quantification des récepteurs internalisés a montré que l’isoforme b internalisait plus efficacement que l’isoforme a suite à l’incubation à la 5-HT (61 ± 3 % pour le b vs 40 ± 2 % pour le a). Les protéines kinases PKA et PKC n’étaient pas impliquées dans cette différence, toutefois, la PKC a été trouvée essentielle à l’internalisation des deux isoformes. L’internalisation de l’isoforme b par 5-HT n’a pas été affecté par la surexpression de forme inactive de GRK2 (GRK2- K220R) et a été partiellement inhibé par un mutant négative de la β-arrestine (βarr(319-418)), tandis que l’internalisation de l’isoforme a a été bloquée par les deux. Ces observations indiquent que les mécanismes d’internalisation des deux isoformes du récepteur 5-HT4 les plus abondants dans le système nerveux central sont distincts. Des comportements spécifiques à chaque isoforme ont aussi été constatés au niveau de la régulation fonctionnelle suite à l’exposition au RS67333, qui désensibilise seulement l’isoforme b. D’après nos observations, nous avons conclu que les isoformes a et b diffèrent dans leur propriétés de signalisation et de régulation. L’incapacité du RS67333 à désensibiliser l’isoforme a fournit un substrat moléculaire pour les effets antidépressifs prolongés de cet agoniste dans les études pré-cliniques. / The therapeutic actions of antidepressants, currently available, require several weeks of treatment. This delay is due to pre-and post-synaptic sites adjustments, which, respectively, increase the availability of synaptic monoamines 5-HT and NA, and induce neuroplastic changes amending neuronal function in the limbic regions. We have recently observed in animal model of depression that serotonergic 4 receptor agonist RS67333 produces behavioural, electrophysiological, cellular and biochemical changes, as seen in antidepressants. More importantly, these changes appear only after 3 days of treatment while antidepressants often require several weeks. Moreover, activation of 5-HT4 receptors does not generate tolerance and that for 21 days of treatment. However, the signalling and regulation properties of these receptors haven’t been established yet. Here, we wanted to better characterize these two aspects of their function, and in particular for isoforms a and b, strongly expressed in the limbic system. First, we assessed their ability to produce cAMP in a heterologous system. Functional assays revealed that both isoforms were capable of positive and negative modulation of cAMP levels by 5-HT. Stimulation by RS67333 induced cAMP stimulation. Together, these observations indicate that that both isoforms are able to couple adenylate cyclase through Gαs and Gαi proteins. Moreover, quantification of receptors sequestration showed that isoform b internalised more efficiently than the isoform a, following incubation with 5-HT (61 ± 3 % for b, 40 ± 2 % for a). PKA and PKC proteins, two seconds messenger kinases activated by these receptors, are not involved in producing this difference. However, PKC is essential to the internalization of both isoforms. Isoform b internalization was not affected by an inactive GRK2 mutant (GRK2-K220R) and was partially inhibited by dominant negative β-arrestin (βarr(319-418), while isoform a internalization was dependant on both. These observations indicate that internalization mechanisms of the two isoforms, most abundantly expressed in the central nervous system, are different. Functional desensitization studies showned that RS67333 selectively desensitizes the isoform b, but not isoforme a. Based on these observations, we conclude that isoform a and b differ in their signalling and regulatory properties. Isoform a incapacity to desensitization by RS67333 provides a molecular substrate for prolonged antidepressant affects of this agonist in pre-clinical studies.