Proprioception is the ability to sense body position necessary for coordinate precise movements. Despite the low complexity of insect neuronal systems, scientists are studying their motor control system. Researchers performed experiments in desert locusts by stimulating their apodeme and recording the neuronal response. Previous studies reported variations in neuronal spiking rates related to acceleration, velocity and position sensitivity. Their results led us to the assumption that either there are different kinds of sensory neurons, or there is only one type of neuron responding to various Physical quantities. Therefore, this research intends to investigate the different spiking rates. We also want to study the influence of apodemes excitations in sensory neurons with information theoretical measures. However, the way signals were recorded does not allow the calculation of delayed transfer entropy (DTE) between sensory neurons. To solve that problem we propose a method to estimate parameters of connections in such scenarios. Our analysis will model the time spent between spikes with survival functions. The influence of excitation in the neuronal response will be analyzed with DTE, which will also be used to validate the methods of simulation. Results show that there is evidence to support the assumption of different spiking rates among sensory neurons. DTE suggests the existence of intermediate processing nodes between excitation and some sensory neurons. A further simulation joining the methods proposed and neuronal signals showed that models considering intermediate pathways present a good fit to the data. We suggest that the different responses of sensory neurons are not due to various types of neurons, but to a preprocessing layer. / Propriocepção é a capacidade de monitorar a posição do corpo necessária para coordenar movimentos precisos. Apesar da baixa complexidade dos sistemas neuronais de insetos, cientistas têm estudado seu controle motor. Pesquisadores realizaram experimentos em gafanhotos estimulando mecanicamente seu apódema e registrando a resposta neuronal. Estudos anteriores relatam variações nas taxas de spiking, e as relacionam com sensibilidades à aceleração, à velocidade e à posição. Seus resultados nos levaram às suposições de que ou existem diferentes tipos de neurônios sensores ou há apenas um tipo de neurônio sensível à diferentes grandezas físicas. Portanto, esta pesquisa pretende investigar as diferentes taxas de spiking e estudar a influência das excitações do apódema em neurônios sensores com medidas de teoria da informação. No entanto, a forma como os sinais foram gravados não permite calcular-se a transferência de entropia atrasada (DTE) entre neurônios sensores. Para tanto, propôs-se um método de estimação de parâmetros para ligações em tais cenários. As análises modelarão o tempo gasto entre spikings com funções de sobrevida. Além disso, a influência da excitação sobre a resposta neuronal será analisada com DTE, a qual também será utilizada para validar os métodos de simulação. Os resultados mostram que há evidências para suportar a hipótese de diferentes taxas de spiking. A DTE sugere a existência de nós intermediários (entre excitação e alguns neurônios sensoriais). Posteriormente, uma simulação juntando os métodos propostos e os sinais neuronais mostrou que modelos considerando caminhos intermediários se ajustam bem aos dados. Por fim, os resultados sugerem que as diferentes respostas de neurônios sensores não acontecem devido a diferentes tipos de neurônios, mas sim à uma camada de pré-processamento.

It is of interest to understand how new neurons incorporate themselves into the existing circuitry of certain neuronal populations. One such population of neurons is that which are born in the subventricular zone (SVZ) and migrate to the olfactory bulb where they differentiate into granule cells. Another area of interest is the role of brain-derived neurotrophic factor (BDNF) on the survival and overall health of these neurons. This study aimed to test whether or not BDNF is a survival factor for adult-born granule cells. Here were utilized a transgenic mouse model over-expressing BDNF under the α- calcium/calmodulin-dependent protein kinase II (CAMKIIα) promoter, and tested its effect on olfactory granule cells under sensory deprived conditions. Results from this experiment indicated that there was no significant difference in cell death or cell survival when comparing transgenic and wild type animals. We concluded that BDNF is not a survival factor for adult-born granule cells. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2016. / FAU Electronic Theses and Dissertations Collection

The effect of interleukin-1 beta (IL-1β) on the electrical properties of sensory neurons was assessed at comparable levels and exposure times to those found in animal models of neuropathic pain. Experiments involved whole cell current- or voltage-clamp recordings from rat dorsal root ganglion (DRG) neurons in defined medium, neuron enriched cultures. 5-6 days exposure to 100 pM IL-1β produced neuron specific effects. These included an increase in the excitability of medium diameter and small diameter isolectin B4 (IB4)-positive neurons that was comparable to that found after peripheral nerve injury. By contrast, a reduction in excitability was observed in large diameter neurons, while no effect was found in small diameter IB4-negative neurons. Further characterization of changes in medium and small IB4-positive neurons revealed that some, but not all, effects of IL-1β were mediated through its receptor, IL-1RI. Using appropriate voltage protocols and/or ion substitutions, it was found that neuron specific changes in several ionic currents, including alterations in hyperpolarization activated inward current (IH) and decreases in various K+ currents contribute to the increased excitability produced by IL-1β. Overall, these studies revealed that: 1. The effects of long-term exposure of DRG neurons to IL-1β are reflective of the enduring increase in primary afferent excitability reported after peripheral nerve injury. This expands the recognized role of IL-1β in acute inflammatory pain to neuropathic pain. 2. Hyperexcitability in medium neurons exposed to IL-1β likely includes mixed populations of neurons corresponding to nociceptive and non-nociceptive primary afferent fibres and, therefore, has relevance to hyperalgesia and allodynia, respectively. 3. The responsiveness of small IB4-positive neurons, but not IB4-negative, to prolonged IL-1β exposure is consistent with the suggestion that small IB4-negative afferents are involved in inflammatory pain, while small IB4-positive afferents are involved neuropathic pain. 4. The identification of receptor mediated effects and several contributing ionic mechanisms, may have relevance to the development of new therapeutic approaches to neuropathic pain. 5. IL-1β can contribute to increased neuronal excitability by mechanisms that are independent of IL-1RI signalling. This should be taken into account when targeting IL-1β, or more specifically IL-1RI, in the management of neuropathic pain.

No description available.

Proprioception is the ability to sense body position necessary for coordinate precise movements. Despite the low complexity of insect neuronal systems, scientists are studying their motor control system. Researchers performed experiments in desert locusts by stimulating their apodeme and recording the neuronal response. Previous studies reported variations in neuronal spiking rates related to acceleration, velocity and position sensitivity. Their results led us to the assumption that either there are different kinds of sensory neurons, or there is only one type of neuron responding to various Physical quantities. Therefore, this research intends to investigate the different spiking rates. We also want to study the influence of apodemes excitations in sensory neurons with information theoretical measures. However, the way signals were recorded does not allow the calculation of delayed transfer entropy (DTE) between sensory neurons. To solve that problem we propose a method to estimate parameters of connections in such scenarios. Our analysis will model the time spent between spikes with survival functions. The influence of excitation in the neuronal response will be analyzed with DTE, which will also be used to validate the methods of simulation. Results show that there is evidence to support the assumption of different spiking rates among sensory neurons. DTE suggests the existence of intermediate processing nodes between excitation and some sensory neurons. A further simulation joining the methods proposed and neuronal signals showed that models considering intermediate pathways present a good fit to the data. We suggest that the different responses of sensory neurons are not due to various types of neurons, but to a preprocessing layer. / Propriocepção é a capacidade de monitorar a posição do corpo necessária para coordenar movimentos precisos. Apesar da baixa complexidade dos sistemas neuronais de insetos, cientistas têm estudado seu controle motor. Pesquisadores realizaram experimentos em gafanhotos estimulando mecanicamente seu apódema e registrando a resposta neuronal. Estudos anteriores relatam variações nas taxas de spiking, e as relacionam com sensibilidades à aceleração, à velocidade e à posição. Seus resultados nos levaram às suposições de que ou existem diferentes tipos de neurônios sensores ou há apenas um tipo de neurônio sensível à diferentes grandezas físicas. Portanto, esta pesquisa pretende investigar as diferentes taxas de spiking e estudar a influência das excitações do apódema em neurônios sensores com medidas de teoria da informação. No entanto, a forma como os sinais foram gravados não permite calcular-se a transferência de entropia atrasada (DTE) entre neurônios sensores. Para tanto, propôs-se um método de estimação de parâmetros para ligações em tais cenários. As análises modelarão o tempo gasto entre spikings com funções de sobrevida. Além disso, a influência da excitação sobre a resposta neuronal será analisada com DTE, a qual também será utilizada para validar os métodos de simulação. Os resultados mostram que há evidências para suportar a hipótese de diferentes taxas de spiking. A DTE sugere a existência de nós intermediários (entre excitação e alguns neurônios sensoriais). Posteriormente, uma simulação juntando os métodos propostos e os sinais neuronais mostrou que modelos considerando caminhos intermediários se ajustam bem aos dados. Por fim, os resultados sugerem que as diferentes respostas de neurônios sensores não acontecem devido a diferentes tipos de neurônios, mas sim à uma camada de pré-processamento.

Les réactions immunitaires de la peau sont initiées par les cellules dendritiques cutanées (dendritic cells, DCs). L'effet potentiellement sensibilisateur d'un composé peut être prédit in vitro en utilisant des monocytes humains différenciés en DCs (MonoDCs). Cependant, ces modèles simplistes restent imprécis car l'activation des DCs cutanés par les sensibilisateurs peut être déclenchée ou modulée par des interactions microenvironnementales avec de multiples types de cellules non immunitaires. Notre objectif est de développer une peau immunocompétente qui combinera des MonoDCs avec tous les éléments structurels et fonctionnels de la peau, c'est-à-dire une barrière épidermique posée sur un derme contenant une pseudo-vascularisation et des neurones nociceptifs. Une matrice de collagène a été ensemencée avec des fibroblastes et des cellules endothéliales, puis avec des précurseurs de fibres nerveuses dérivées soit de l'iPSC humaine, soit de la DRG embryonnaire murins. Enfin, nous avons introduit les MonoDC et les kératinocytes. Nous avons observé que les neurones différenciés in situ innervent l'épiderme comme observé habituellement dans la peau humaine normale. De plus, les neurones dérivées d’iPSCs, expriment neuropeptides et canaux calcique spécifiques des fibres nociceptives. Enfin, les Mono-DC intégrés au modèle restent stable pendant toute la durée nécessaire à la formation de l’épiderme et peuvent être stimulé. Le modèle sera utilisé pour prédire le potentiel irritant des composés chimiques et l'impact de l’innervation nociceptive sur l'activation des DCs. / Immune reactions in the skin are initiated by the cutaneous dendritic cells (DCs). The potential sensitizing effect of a compound can be predicted in vitro using human monocytes differentiated into DCs (Mono-DCs). However, these simplistic models remain inaccurate because the activation of cutaneous DCs by sensitizers may be triggered or modulated by microenvironmental interactions with multiple types of non-immune cells. Our goal is to develop an immunocompetent human tissue-engineered skin that will combine DCs with all structural and functional element of the skin, i.e. an epidermal barrier laid upon a dermis containing a pseudo-vascularization and nociceptive neurons. Collagen matrix was seeded with fibroblasts and endothelial cells, then with precursors of nerve fibers derived from either human iPSC or murine embryonic DRG. Finally, we introduced Mono-DCs and keratinocytes. We observed that in situ differentiated neurons grow axons towards the epidermis as usually observed in normal human skin. What's more, the neurons derive from iPSC, express neuropeptides and calcium channel as normal nociceptive fibers. Moreover, Mono-DCs settled as expected beneath the epidermis and remained sessile to stimulation for several weeks. The model will be used to predict the irritant potential of chemical compounds, and the impact of nerves on DC activation.

Philosophiae Doctor - PhD / Tsetse flies are the biological vectors of human and animal trypanosomiasis and hence representant medical and veterinary importance. The sense of smell plays a significant role in tsetse and its ecological interaction, such as finding blood meal source, resting, and larvicidal sites and for mating. Tsetse olfactory behaviour can be exploited for their management; however, olfactory studies in tsetse flies are still fragmentary. Here in my PhD thesis, using scanning electron microscopy, electrophysiology, behaviour, bioinformatics and molecular biology techniques, I have investigated tsetse flies (Glossina fuscipes fuscipes) olfaction using behaviourally well studied odorants, tsetse repellent by comparing with attractant odour. Insect olfaction is mediated by olfactory sensory neurons (OSNs), located in olfactory sensilla, which are cuticular structures exposed to the environment through pore and create a platform for chemical communication. In the sensilla shaft the dendrite of OSNs are housed, which are protected by called the sensillum lymph produced by support cells and contains a variety of olfactory proteins, including the odorant binding protein (OBP) and chemosensory proteins (CSP). While on the dendrite of OSNs are expressed olfactory receptors. In my PhD, studies I tried to decipher the sense of smell in tsetse fly. In the second chapter, I demonstrated that G. f. fuscipes is equipped with diverse olfactory sensilla, that various from basiconic, trichoid and coeloconic. I also demonstrated, there is shape, length, number difference between sensilla types and sexual dimorphism. There is a major difference between male and female, while male has the unique basiconic sensilla, club shaped found in the pits, which is absent from female pits. In my third chapter, I investigated the odorant receptors which are expressed on the dendrite of the olfactory sensory neurons (OSNs). G. f. fuscipes has 42 ORs, which were not functionally characterised. I used behaviourally well studied odorants, tsetse repellents, composed of four components blend. I demonstrated that tsetse repellent is also a strong antifeedant for both G. pallidipes and G. f. fuscipes using feeding bioassays as compared to the attractant odour, adding the value of tsetse repellent. However, the attractant odour enhanced the feeding index. Using DREAM (deorphanization of receptors based on expression alterations of mRNA levels). I found that in G. f. fuscipes, following a short in vivo exposure to the individual tsetse repellent component as well as an attractant volatile chemical, OSNs that respond to these compounds altered their mRNA expression in two opposite direction, significant downregulation and upregulation in their number of transcripts corresponding to the OR that they expressed and interacted with odorant. Also, I found that the odorants with opposite valence already segregate distinctly at the cellular and molecular target at the periphery, which is the reception of odorants by OSNs, which is the basis of sophisticated olfactory behaviour. Deorphanization of ORs in none model insect is a challenge, here by combining DREAM with molecular dynamics, as docking score, physiology and homology modelling with Drosophila a well-studied model insects, I was able to predict putative receptors of the tsetse repellent components and an attractant odour. However, many ORs were neutral, showing they were not activated by the odorants, demonstrating the selectivity of the technique as well as the receptors. In my fourth chapter, I investigated the OBPs structures and their interaction with odorants molecules. I demonstrated that OBPs are expressed both in the antenna, as well as in other tissues, such as legs. I also demonstrated that there are variations in the expression of OBPs between tissues as well as sexes. I also demonstrated that odorants induced a fast alteration in OBP mRNA expression, some odorants induced a decrease in the transcription of genes corresponding to the activated OBP and others increased the expression by many fold in OBPs in live insect, others were neutral after 5 hours of exposure. Moreover, with subsequent behavioural data showed that the behavioural response of G. f. fuscipes toward 1-octen-3-ol decreased significantly when 1-octen-3-ol putative OBPs were silenced with feeding of double-stranded RNA (dsRNA). In summary, our finding whereby odorant exposure affects the OBPs mRNA, their physiochemical properties and the silencing of these OBPs affected the behavioural response demonstrate that the OBPs are involved in odour detection that affect the percept of the given odorant. The expression of OBPs in olfactory tissues, antenna and their interaction with odorant and their effect on behavioural response when silenced shows their direct involvement in odour detection and reception. Furthermore, their expression in other tissues such as legs indicates they might also have role in other physiological functions, such as taste.

No description available.

No description available.

Philosophiae Doctor - PhD / Tsetse flies are the biological vectors of human and animal trypanosomiasis and hence representant medical and veterinary importance. The sense of smell plays a significant role in tsetse and its ecological interaction, such as finding blood meal source, resting, and larvicidal sites and for mating. Tsetse olfactory behaviour can be exploited for their management; however, olfactory studies in tsetse flies are still fragmentary. Here in my PhD thesis, using scanning electron microscopy, electrophysiology, behaviour, bioinformatics and molecular biology techniques, I have investigated tsetse flies (Glossina fuscipes fuscipes) olfaction using behaviourally well studied odorants, tsetse repellent by comparing with attractant odour. Insect olfaction is mediated by olfactory sensory neurons (OSNs), located in olfactory sensilla, which are cuticular structures exposed to the environment through pore and create a platform for chemical communication. In the sensilla shaft the dendrite of OSNs are housed, which are protected by called the sensillum lymph produced by support cells and contains a variety of olfactory proteins, including the odorant binding protein (OBP) and chemosensory proteins (CSP). While on the dendrite of OSNs are expressed olfactory receptors. In my PhD, studies I tried to decipher the sense of smell in tsetse fly. In the second chapter, I demonstrated that G. f. fuscipes is equipped with diverse olfactory sensilla, that various from basiconic, trichoid and coeloconic. I also demonstrated, there is shape, length, number difference between sensilla types and sexual dimorphism. There is a major difference between male and female, while male has the unique basiconic sensilla, club shaped found in the pits, which is absent from female pits. In my third chapter, I investigated the odorant receptors which are expressed on the dendrite of the olfactory sensory neurons (OSNs). G. f. fuscipes has 42 ORs, which were not functionally characterised. I used behaviourally well studied odorants, tsetse repellents, composed of four components blend. I demonstrated that tsetse repellent is also a strong antifeedant for both G. pallidipes and G. f. fuscipes using feeding bioassays as compared to the attractant odour, adding the value of tsetse repellent. However, the attractant odour enhanced the feeding index. Using DREAM (deorphanization of receptors based on expression alterations of mRNA levels). I found that in G. f. fuscipes, following a short in vivo exposure to the individual tsetse repellent component as well as an attractant volatile chemical, OSNs that respond to these compounds altered their mRNA expression in two opposite direction, significant downregulation and upregulation in their number of transcripts corresponding to the OR that they expressed and interacted with odorant. Also, I found that the odorants with opposite valence already segregate distinctly at the cellular and molecular target at the periphery, which is the reception of odorants by OSNs, which is the basis of sophisticated olfactory behaviour. Deorphanization of ORs in none model insect is a challenge, here by combining DREAM with molecular dynamics, as docking score, physiology and homology modelling with Drosophila a well-studied model insects, I was able to predict putative receptors of the tsetse repellent components and an attractant odour. However, many ORs were neutral, showing they were not activated by the odorants, demonstrating the selectivity of the technique as well as the receptors. In my fourth chapter, I investigated the OBPs structures and their interaction with odorants molecules. I demonstrated that OBPs are expressed both in the antenna, as well as in other tissues, such as legs. I also demonstrated that there are variations in the expression of OBPs between tissues as well as sexes. I also demonstrated that odorants induced a fast alteration in OBP mRNA expression, some odorants induced a decrease in the transcription of genes corresponding to the activated OBP and others increased the expression by many fold in OBPs in live insect, others were neutral after 5 hours of exposure. Moreover, with subsequent behavioural data showed that the behavioural response of G. f. fuscipes toward 1-octen-3-ol decreased significantly when 1-octen-3-ol putative OBPs were silenced with feeding of double-stranded RNA (dsRNA). In summary, our finding whereby odorant exposure affects the OBPs mRNA, their physiochemical properties and the silencing of these OBPs affected the behavioural response demonstrate that the OBPs are involved in odour detection that affect the percept of the given odorant. The expression of OBPs in olfactory tissues, antenna and their interaction with odorant and their effect on behavioural response when silenced shows their direct involvement in odour detection and reception. Furthermore, their expression in other tissues such as legs indicates they might also have role in other physiological functions, such as taste.