These studies compare neurotransmitter localization and the behavioral functions of homologous neurons in nudibranch molluscs to determine the types of changes that might underlie the evolution of species-specific behaviors. Serotonin (5-HT) immunohistochemistry in eleven nudibranch species indicated that certain groups of 5 HT-immunoreactive neurons, such as the Cerebral Serotonergic Posterior (CeSP) cluster, are present in all species. However, the locations and numbers of many other 5 HT-immunoreactive neurons were variable. Thus, particular parts of the serotonergic system have changed during the evolution of nudibranchs. To test whether the functions of homologous neurons are phylogenetically variable, comparisons were made in species with divergent behaviors. In Tritonia diomedea, which crawls and also swims via dorsal-ventral body flexions, the CeSP cluster includes the Dorsal Swim Interneurons (DSIs). It was previously shown that the DSIs are members of the swim central pattern generator (CPG); they are rhythmically active during swimming and, along with their neurotransmitter 5-HT, are necessary and sufficient for swimming. It was also known that the DSIs excite efferent neurons used in crawling. DSI homologues, the CeSP-A neurons, were identified in six species that do not exhibit dorsal-ventral swimming. Many physiological characteristics, including excitation of putative crawling neurons were conserved, but the CeSP A neurons were not rhythmically active in any of the six species. In the lateral flexion swimmer, Melibe leonina, the CeSP-A neurons and 5-HT, were sufficient, but not necessary, for swimming. Thus, homologous neurons, and their neurotransmitter, have functionally diverged in species with different behaviors. Homologous neurons in species with similar behaviors also exhibited functional divergence. Like Melibe, Dendronotus iris is a lateral flexion swimmer. Swim interneuron 1 (Si1) is in the Melibe swim CPG. However, its putative homologue in Dendronotus, the Cerebral Posterior ipsilateral Pedal (CPiP) neuron, was not rhythmically active during swim-like motor patterns, but could initiate such a motor pattern. Together, these studies suggest that neurons have changed their functional relationships to neural circuits during the evolution of species-specific behaviors and have functionally diverged even in species that exhibit similar behaviors.

À partir des ovocytes de la palourde Spisula solidissima, un ADNc codant un récepteur nommé Spi-OAR a été cloné et séquencé. Une analyse de la séquence en acides aminés a indiqué que ce nouveau récepteur possède une forte similarité avec les récepteurs β-adrénergiques et les récepteurs à octopamine. En effet, il est étroitement lié à la classe des récepteurs à octopamine « β-adrénergique-like » couplés à une protéine Gs. L’ADNc de Spi-OAR a été introduit dans un vecteur d'expression (pCEP4) et un épitope reconnaissable par un anticorps commercial a été ajouté au segment N-terminal. Cette construction a été transfectée dans des cellules hôtes (HEK 293) et des études d’immunofluorescence ont montré une expression efficace du récepteur au niveau membranaire. Également, des mesures d'AMPc pour les cellules exprimant Spi-OAR ont révélé une augmentation de ce messager secondaire lors de l'ajout de l'octopamine, et dans une moindre mesure, la tyramine, tandis que la dopamine, la sérotonine et l'histamine n’ont engendré aucun effet. Une légère activité constitutive de ce récepteur dans les cellules hôtes a été observée. De plus, une analyse RT-PCR avec des oligonucléotides spécifiques a révélé l'ARNm de Spi-OAR non seulement dans les ovocytes, mais aussi dans les gonades, le cœur, les muscles adducteurs, les branchies et les ganglions suggérant que ce récepteur soit exprimé de façon ubiquitaire dans divers tissus et dans différents stades embryonnaires chez la palourde. En outre, des études avec des ovocytes isolés n'ont montré aucun effet de l’octopamine sur la réactivation méiotique. Des études éventuelles pourront finalement confirmer le rôle fonctionnel de Spi-OAR. / A cDNA encoding for an octopamine receptor named Spi-OAR was cloned and sequenced from the surf clam Spisula solidissima oocytes. An analysis of its predicted amino acid sequence showed a high degree of similarity with β-adrenergic and octopamine receptors. This receptor qualifies as a novel receptor closely related to the proposed class of insect octopamine « β-adrenergic–like » receptors coupled to Gs protein. This cDNA was introduced into an expression vector (pCEP4), with an added N-terminal FLAG tag sequence, and transfected in host cells (HEK 293). Immunofluorescence studies showed expression of the receptor with a proper localization to the plasma membrane. Measurements of cAMP in transfected cells revealed that addition of octopamine, and to a lower extent, tyramine induced a rise in cAMP while dopamine, serotonine and histamine had no effect. Overexpression of Spi-OAR in mammalian cells induced slight constitutive increase of cAMP. An RT-PCR analysis with specific oligonucleotides revealed the presence of the receptor mRNA not only in oocytes but also in whole gonads, heart, adductor muscle, gills and ganglia suggesting that this receptor is likely ubiquitously expressed. Expression of Spi-OAR was also detected at different embryonic stages. Despite the demonstrated expression of Spi-OAR in oocytes, octopamine had no effect on meiotic reinitiation. Further studies will examine the function of Spi-OAR.

Les activités humaines font peser sur l’écosystème marin de multiples pressions délétères. Pollution chimique, changement climatique, risque d’acidification, débris de plastique et déchets radioactifs ont des impacts sans précèdent. Une pollution de plus en plus reconnu comme majeur est la pollution sonore. La prospection sismique, le battage de pieux et le trafic maritime génèrent des niveaux sonores qui peuvent être extrêmement forts, modifiant fondamentalement le paysage acoustique sous-marin. On sait que de nombreux mammifères marins et poissons entendent le bruit généré par ces activités et que cela altère leur physiologie et leur éthologie. Par contre, chez les invertébrés marins très peu d’études avaient évalué leur capacité à entendre et l’impact de cette pollution sur eux reste à déterminer. Nous avons abordé le problème par une étude de la capacité de perception du son chez l'huître creuse Magallana gigas en utilisant une approche comportementale et physiologique. Nous avons montré que M. gigas entend dans la gamme de fréquences entre 10 et 1000 Hz. Cette analyse nous a permis de caractériser les sources de sons qui contribuent à leur environnement auditif. Au laboratoire, dans un milieu contaminé (i) au cadmium, un métal que nous avons considéré à la fois comme une substance toxique et un marqueur indirect de l'activité ventilatoire, et (ii), par des bruits de cargo, nous montrons un effet répresseur du bruit caractérisé par une diminution de l'activité valvaire, de l'activité ventilatoire et du taux de croissance. Nous rapportons également une diminution de la bioaccumulation du Cd dans les branchies et une modulation de l'expression de certains gènes. Nous avons enfin étudié sur un enregistrement de 2 ans dans le port commercial de Santander, le comportement (incluant les pontes et la croissance) de 3 groupes d’huitres exposés à une forte pollution sonore et à une « qualité de l’eau » considérée dans la littérature comme bonne à très bonne pour une masse d’eau fortement modifiée. Nous avons retrouvé dans notre analyse différents effets que nous avions provoqués ou prédits à partir du travail de laboratoire où nous avions manipulé le bruit seul. Nous concluons que la pollution sonore au sein du port doit diminuer le fitness des huîtres en modifiant leur activité valvaire, la hiérarchie de leurs rythmes biologiques et la croissance. Nos résultats suggèrent que la pollution sonore peut avoir des conséquences importantes sur les invertébrés et présente un risque fort en termes de productivité de l'écosystème. / Human activities introduce multiple harmful pressures on the marine ecosystem. Chemical pollution, climate change, acidification risk, plastic debris and radioactive wastes have significant effects on marine wildlife. Noise pollution is now recognized as a major source of pollution at sea. Seismic exploration, pile driving and marine traffic, among other activities, generate noise at high sound pressure levels altering the underwater acoustic landscape. Many marine mammals and fish hear the noise generated by these activities which have the potential to alter their physiology and ethology. However, very few studies among marine invertebrates had assessed their ability to hear and the impact of noise pollution on them has yet to be determined. We approached the problem by studying sound perception ability in the pacific oyster Magallana gigas using behavioural and physiological techniques. We have shown that M. gigas is sensitive to sound in the frequency range from 10 to 1000 Hz. This characterization allowed us to define sound sources that contribute to their sound landscape. In the laboratory, in an environment contaminated with (i) cadmium, a metal that we considered to be both a toxic agent and an indirect marker of ventilatory activity, and (ii) cargo ship noise, we showed a depressant or repressant effect of noise characterized by a decrease in valve activity, ventilatory activity and growth rate. We also report a decrease in Cd bioaccumulation and some modulation of gene expression. Finally, we studied a 2-year behavioural record performed in the commercial port of Santander (including spawning events and growth) on 3 groups of oysters exposed to high noise pressure levels. In the port of Santander, the "water quality" is otherwise considered by the literature as good to very good for a heavily modified water body. We found in these records different changes that we previously induced and/or produced in the laboratory. We conclude that the noise pollution load occurring within a commercial port must reduce the fitness of oysters by modifying their valve activity, the hierarchy of their biological rhythms and their growth rate. Our results strongly suggest that noise pollution can have significant consequences on invertebrates and presents a high risk in terms of ecosystem productivity.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-05-12T19:07:27Z No. of bitstreams: 1 viniciusjosepilate.pdf: 783213 bytes, checksum: da2d70d6a4638a79930055e8f708be6c (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-06-27T20:07:20Z (GMT) No. of bitstreams: 1 viniciusjosepilate.pdf: 783213 bytes, checksum: da2d70d6a4638a79930055e8f708be6c (MD5) / Made available in DSpace on 2016-06-27T20:07:20Z (GMT). No. of bitstreams: 1 viniciusjosepilate.pdf: 783213 bytes, checksum: da2d70d6a4638a79930055e8f708be6c (MD5) Previous issue date: 2013-02-27 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O conhecimento dos aspectos biológicos, morfológicos e comportamentais dos moluscos terrestres é importante para o desenvolvimento de medidas de controle de pragas e de estratégias de conservação de espécies. O presente trabalho teve como objetivo caracterizar os padrões de biologia, conquiliomorfometria e comportamento da espécie Dysopeas muibum e verificar os efeitos do isolamento e do fotoperíodo sobre seu ciclo de vida e a morfometria da concha, em condições de laboratório. Foi realizado o acompanhamento do crescimento da concha, da liberação de filhotes e da mortalidade de 160 moluscos, dos quais 40 foram mantidos isolados e 120 agrupados (10 moluscos por grupo; 40 em condições naturais de temperatura, umidade relativa do ar e fotoperíodo, e 80 tendo estas condições controladas – 40 com dias longos e 40 com dias curtos), desde o nascimento até 180 dias de vida, quando então foram aferidas as medidas da concha. Para verificar o horário de atividade e compor o etograma, 30 moluscos adultos (10 moluscos por grupo) foram observados durante 24 horas contínuas, através do método de varredura, com registro dos atos comportamentais a intervalos de 20 minutos. Já para verificar a ocorrência de agregação e a preferência por sítio de repouso, durante cinco dias, a intervalos de 24 horas, foram observados de 30 moluscos adultos (10 moluscos por grupo): número de agregações, de indivíduos agregados e isolados, e de indivíduos sob o disco com alimento, sob o disco sem alimento, sobre o substrato, enterrados e na parede do terrário. Para todas as análises, os moluscos foram criados em terrários plásticos vedados com tecido de algodão e elástico, contendo como substrato terra vegetal esterilizada e umedecida com água de torneira, e alimentados com ração para frangos de corte enriquecida com carbonato de cálcio. Foi verificado que a espécie ocorre no município de Juiz de Fora, Minas Gerais, possui parte mole amarelada e concha pequena e alongada, e é capaz de se reproduzir por autofecundação. Possui crescimento indeterminado, fecundidade aumentando com o tamanho corporal, iteroparidade e baixa mortalidade antes e após a maturidade sexual, não tendo o fotoperíodo efeito sobre esses padrões. As condições controladas de temperatura, umidade relativa do ar e fotoperíodo favoreceram o crescimento, maiores valores conquiliomorfométricos e o rápido alcance da maturidade sexual, enquanto a diminuição do comprimento do dia favoreceu a fecundidade. O isolamento aumentou o número total de filhotes, o número de filhotes por molusco parental e o intervalo entre eventos reprodutivos, reduziu o número de eventos reprodutivos e de filhotes por evento, e não influenciou o crescimento, o alcance da maturidade sexual, a relação crescimento-reprodução, a mortalidade e a conquiliomorfometria. O etograma foi composto por oito atos comportamentais: repousar, estar enterrado, deslocar, enterrar, explorar, alimentar, emergir e interagir. A espécie apresentou maior tendência ao repouso e a locomoção foi o ato comportamental de atividade prevalente, não apresentou horário de atividade predominantemente noturno e comportamento agregativo, e seu sítio preferido para o repouso foi o enterramento no substrato. / Knowledge of biological, morphological and behavioral characteristics of terrestrial molluscs is important for the development of pest control measures and strategies for the conservation of species. This study aimed to characterize the patterns of biology, conchiliomorphometric and behavior of the species Dysopeas muibum and check the effects of isolation and photoperiod on its life cycle and shell morphometry, under laboratory conditions. We conducted monitoring growth of the shell, release of puppies and mortality of 160 molluscs, of which 40 were kept isolated and 120 grouped (10 molluscs per group; 40 in natural conditions of temperature, relative humidity and photoperiod, and 80 with controlled conditions – 40 with long days and 40 with short days), from birth to 180 days of life, when the measures of the shell were taken. To check the schedule of activity and compose the ethogram, 30 adult molluscs (10 molluscs per group) were observed for 24 continuous hours, through the scanning method, with a record of behavioral acts at intervals of 20 minutes. For verifying the occurrence of aggregation and the preference for resting place, for five days, at 24 hour intervals, were observed in 30 adult molluscs (10 molluscs per group): number of aggregations, of aggregates and isolated individuals, and of individuals below the disc with food, below the disc without food, on the substrate, buried and in the wall of terrarium. For all analyzes, the molluscs were reared in plastic terrariums sealed with cotton fabric and elastic, containing plant soil sterile and moistened with tap water, and fed with food to broilers enriched with calcium carbonate. It was found that the species occurs in the Juiz de Fora City, MG, Brazil, has the soft yellowish and small and elongated shell, and is able to reproduce by self-fertilization. It has indeterminate growth, fecundity increases with body size, iteroparity and low mortality before and after sexual maturity, not having the photoperiod effect on these patterns. The controlled conditions of temperature, relative humidity and photoperiod favored growth, conchiliomorphometric values higher and fast reach sexual maturity, while the decrease in day length favored fecundity. The isolation increased the total number of pups, number of pups per parent mollusc and the interval between reproductive events, reduced the number of reproductive events and pups per event, and did not influence growth, reach sexual maturity, growth-reproduction relation, mortality and conchiliomorphometric. The ethogram was composed of eight behavioral acts: lie, lie buried, move, bury, explore, food, emerge and interact. The species had a greater tendency to rest and locomotion was the behavioral act prevalent activity, showed no activity time predominantly nocturnal and affiliative behavior, and their favorite place to rest was buried in substrate.

Denna studien har gjorts på blåmusslor, Mytilus edulis trossulus, för att undersöka effekter av det antidepressiva läkemedlet Citalopram på blåmusslans fysiologiska funktioner. Blåmusslan är en vanligt förekommande art i Östersjön där den fungerar som en nyckelart som; bidrar till den biologiska mångfalden, är långt ner i näringskedjan och är anpassningsbar. Dessa egenskaper gör den även lämpad som testorganism i denna studie. Citalopram är ett av Sveriges vanligaste läkemedel för att behandla psykisk ohälsa och den verkar genom att blockera återupptaget av serotonin i hjärnan och ingår i läkemedelsgruppen serotoninhämmare (SSRI). Serotonin påverkar flera olika funktioner i kroppen såsom rörelseförmåga, sinnesstämningar och mättnadskänsla. Läkemedel inklusive Citalopram återfinns i sjöar och vattendrag, detta för att läkemedel inte renas bort i våra reningsverk och för att det idag inte finns några krav på läkemedelsrening i svenska avloppsreningsverk. Läkemedel tillverkas för att vara aktiva vid låga koncentrationer vilket gör att de även har en negativ effekt på marina organismer. I denna studie har vi analyserat effekter av Citalopram på blåmusslans fysiologiska funktioner genom tre analyser, filtrering, rörelse och produktion av byssustrådar i tre olika koncentrationer (0 ng/l, 20 ng/l och 200 ng/l). Analyserna utfördes under två tidsperioder, exponering av läkemedel under 96 timmar samt remediering utan läkemedel under 72 timmar. Studien visar en signifikant skillnad i filtrering (P-värde = 0.04416) mellan grupperna under exponeringen där fler blåmusslor som var exponerade för 200 ng/l visade en lägre filtreringsaktivitet (fler individer var helt stängda). Under remedieringen observerades en signifikant högre mortalitet bland blåmusslor som exponerats för 200 ng/l (P-värde = 0.01347). Blåmusslor exponerade för 200 ng/l Citalopram visade även en lägre produktion av byssustrådar (P-värde= 0.02702). I den här studien kan vi se att läkemedlet Citalopram har en negativ påverkan på flera av blåmusslans fysiologiska funktioner. / The aim of this study was to investigate the effects of the antidepressant drug Citalopram on blue mussels, Mytilus edulis trossulus, in the Baltic Sea. The blue mussel is a common species that is considered a key species in the Baltic Sea as it contributes to biodiversity, and is adaptable which makes it suitable as a test organism in this study. Selective serotonin reuptake inhibitors (SSRIs) are the most commonly prescribed antidepressants and a common SSRI substance is Citalopram. This substance is one of Sweden's most common drugs for treating mental illness. Serotonin affects several different functions in the body such as movement, mood and saturation. Many pharmaceuticals including Citalopram are found in lakes and streams, this is because pharmaceuticals are not purified in our wastewater treatment plants and there is no requirements for filtration of pharmaceuticals in Swedish wastewater treatment plants as of today. Pharmaceuticals are manufactured to be active even at low concentrations, a negative side effect of this is that it is also active in the environment causing negative effects on marine organisms. In this study, we analyzed the effects of Citalopram on the physiological functions of the blue mussel by three analyzes; filtration, movement and production of byssus threads using three different concentrations (0 ng /l, 20 ng/l och 200 ng/l) of Citalopram. The analyzes were carried out over two time periods, exposure of drugs for 96 hours and remediation without drugs for 72 hours. The study shows a significant difference in filtration (P-value = 0.04416) between the groups during exposure, where more blue mussels that were exposed to 200 ng/l showed a lower filtration activity (more individuals completely closed). During the remediation, a significantly higher mortality was observed among blue mussels exposed to 200 ng/l (P value = 0.01347). Blue mussels exposed to 200 ng/l Citalopram also showed lower production of byssus threads (P value = 0.02702). In this study we can see that the drug Citalopram has a negative effect on several of the blue mussel's physiological functions.

The Permian Meade Peak Member of the Phosphoria Formation was examined at four localities in this investigation. Fossils were collected at each locality. The four collecting localities visited in this investigation are: Brazier Canyon, Utah, Montpelier Canyon, Idaho, Coal Canyon, Wyoming, and Cokeville, Wyoming. The environment of deposition of the Meade Peak Member in the study area is inferred to have been an outer shelf or basin characterized by moderately deep and quiet water. This has aided reconstruction of life habits, population dynamics, and growth characteristics of the examined species. The articulate brachiopod Leiorhynchoidea weeksi is inferred to have attached itself to the substrate by means of a pedicle. The observed variability in the sulcus of this species is assumed to have been influenced by intraspecific competition, which may reflect partitioning of nutrient resources at different levels in the water above the substrate. The articulate brachiopod Lissochonetes ostiolatus probably assumed an unattached existence, lying free on the substrate. Several population samples exhibited stunted growth relative to the L. ostiolatus population sample from Montpelier Canyon. Two environmental variants possibly contributed to the inhibition of growth. These are negative Eh, inferred from the associated organic matter in the lithologies, and competition for space. Lingula carbonaria an inarticulate brachiopod, is thought to have had an infaunal mode of life. A population sample of L. carbonaria from Cokeville exhibits substantially larger morphologic mean sizes than two other population samples. These differences can be explained by the fact the sandy lithology from which the larger collection was taken was associated with conditions which facilitated growth. Also, availability of phosphate, inferred from the P2O5 content of the lithology, probably accelerated growth because inarticulates utilize calcium phosphate as shell material. Orbiculoidea missouriensis is an inarticulate brachiopod. It is inferred to have been attached to the substrate by a pedicle. OF the three population samples of O. missouriensis analyzed, the sample from Brazier Canyon displays larger morphologic mean sizes. This is interpreted as indicating that reducing conditions, inferred from associated organic matter, were milder in that environment. The gastropod bablyonites ferrieri displaysa a low, expanded from, and is thought to have crawled over the surface of the sediment. The food source of this gastropod is not known, but may have been algae, carrion, detritus, or soft-bodied invertebrates. Two bivalves collected are assumed to have been shallow burrowing, labial palp feeders. Both Nuculopsis montpelierensis and polidevcia obesa represent this mode of life. Of the two population samples of P. obesa examined, the one from Cokeville displays larger morphologic means. The environmental stimuli proposed for this difference is competition and an inferred softer substrate in the montpelier assemblage. The bivalve Edmondia phosphatica is inferred to have been a shallow burrowing, siphonate species. Streblochondria montpelierensis and Aviculopecten phosphaticus are bivalves related to modern pectenoids. S. montpelierensis is assumed to have been a byssally attached epifaunal bivalve. The environmental factors influencing the morphologic variation are also inferred to have affected size-frequency distributions and survivorship curves. The effects of reducing conditions have resulted in higher juvenille mortality and/or inhibited growth. Coarser substrates have yielded population samples that exhibit negatively skewed distributions, whereas finer-grained substrates have yielded positively skewed distributions. Growth curves constructed for each population sample generally show a decline in growth rate with age. Some linear trends are noted. Growth lines were used to infer an approximate life span for each species. Conservative approximations of the life spans of the examined species are: L. weeksi, 9 years; L. ostiolatus, 5 years; L. carbonaria, 8 years; A. phosphaticus, 4 years; O. missouriensis, 2.3 years; P. obesa, 5 years; and B. ferrieri, 3.4 years.

Chemosensation is achieved through the binding of chemical signals to chemoreceptor proteins embedded in the membranes of sensory neurons. The molecular identity of these receptors, as well as the downstream processing of chemosensory signals, has been well studied in arthropods and vertebrates. However, very little is known about molluscan chemosensation. The identity of chemoreceptor proteins in the nudibranch mollusc Berghia stephanieae are unknown. Data from other protostome and molluscan studies suggest Berghia may use ionotropic receptors for some forms of chemoreception. This study used a bioinformatics approach to identify potential chemosensory ionotropic receptors in the transcriptome of Berghia. A hidden Markov model program was used to generate molecular profiles of previously identified chemosensory receptors in other animals. A Berghia transcriptome was then searched for likely homologous sequences. Candidate sequences were investigated using protein prediction tools and molecular phylogenies. Fourteen ionotropic glutamate receptors (likely synaptic) and five divergent ionotropic receptors were identified. One of these divergent ionotropic receptor sequences, IR-D, may encode a chemosensory receptor and was therefore selected to determine its cellular expression in sensory and brain tissue using in situ hybridization chain reaction. Expression was seen in the rhinophores and oral tentacles of Berghia, as well as in the rhinophore ganglion, cerebral-pleural ganglion, and pedal ganglion. Similar expression patterns were obtained with tissue-specific transcriptomic data. This was the first study to investigate IR-D as a potential chemosensory receptor in molluscs, and thus has helped identify a new family of possible ionotropic chemoreceptor proteins in molluscs. These results have laid the groundwork for continued investigation of Berghia’s chemosensory system.

La science a avancé depuis le XIX ème siècle. Des nouveaux outils sont apparus : la microscopie optique nous donne la vision des cellules, la microscopie électronique nous entraine au cœur de celles-ci. L’imagerie par résonance magnétique est apparue dans les années soixante-dix. Depuis son évolution, l’IRM nous entraine de plus en plus loin dans les profondeurs secrètes de nos cerveaux. La possibilité d’observer l’activité neuronale à l’aide de l’imagerie fonctionnelle est une grande révolution. Cette thèse montrera la possibilité que l’on a d’observer l’activité d’un neurone individuel sans modification de son réseau grâce à l’imagerie rehaussée au manganèse. L’étude sera effectuée sur des Aplysies à très haut champ (17T). Ces animaux sont des mollusques marins gastropodes qui possèdent une particularité : leurs neurones sont de tailles importantes, ils peuvent atteindre 1 mm de diamètre. Leurs neurones sont regroupés en plusieurs ganglions. Mon étude portera sur le ganglion buccal, qui est le ganglion le plus étudié dans les recherches en électrophysiologie. Avant de réaliser les acquisitions, j’ai dû concevoir plusieurs antennes de tailles microscopiques adaptées à la taille des ganglions. En réduisant la taille des antennes, le rapport signal sur bruit augmente. Dans un deuxième temps, une double antenne a été développée permettant l’acquisition de deux échantillons simultanément. Cette antenne a nécessité de créer des préamplificateurs fonctionnant à 730 MHz. La première série d’expériences a permis d’observer l’évolution de l’activité neuronale selon différents stimulus liés au comportement alimentaire des aplysies in-vivo. J’ai montré grâce à la technique mise en place que l’on peut distinguer par IRM l’activité de chaque neurone face à un stimulus. Par la suite, pour continuer ce travail, une deuxième série d’expériences a été effectuée in-vitro. J’ai étudié le comportement des neurones selon les neuro-stimulateurs perfusés : la dopamine et la sérotonine, tous les deux présents naturellement dans l’aplysie. Globalement les neurones ont été activés mais après les avoir observés individuellement, j’ai remarqué quelques différences selon les neurotransmetteurs. Cette technique peut maintenant être utilisée pour étudier d’autres conduites de l’aplysie comme le comportement compulsif. L’étude sur la mémoire peut être aussi envisagée. Les origines comportementales ont probablement des mécanismes identiques entre les différentes espèces animales et donc avec l’Homme comme l’a démontré les études d’Eric Kandel sur la mémoire. / Science has evolved since the 19th century. New tools have appeared such as optical microscopy which gives us the vision of cells and electronic microscopy which leads us into their hearts. The magnetic resonance imaging appeared in the seventies. Evolving over time, the MRI has taken us farther and farther into the secret depths of our brains. The possibility of observing the neuronal activity thanks to the functional imaging is a major evolution. This thesis will show the possibility we have to observe the activity of a single neuron without modification of its network thanks to the manganese enhanced magnetic resonance imaging technique. The study was done on the Aplysia at very high field magnet (17T). These animals are marine gastropod mollusks with a peculiarity: their neurons are of important size and can reach 1 mm in diameter. Their neurons are grouped into several ganglia. My study concerns the buccal ganglion which is the most studied ganglia in the research in electrophysiology. Before making any acquisitions, I had to conceive several microscopic coils adapted to the size of the ganglions. By reducing the size of the coils, the signal of the noise ratio increases. Then, a double coil allowing the simultaneous acquisition of two samples was built. This antenna required the construction of pre-amplifiers operating at 730 MHz. The first series of experiments helped observe the evolution of the neuronal activity according to different stimuli linked to the eating habits of the Aplysia in vivo. Thanks to the technique implemented, I shall show that, using MRI, it is possible to distinguish the activity of each neuron with respect to a stimulus. Afterwards, to continue this work, a second series of experiments was made in vitro. I studied the behavior of neurons when perfused with neural stimulators: dopamine and serotonin, both naturally present in the Aplysia. Generally, all neurons were activated but when observing them individually, I noticed some differences. Studies in electrophysiology will allow us to get a better understanding and a confirmation of the results of this study. The MEMRI technique can be used in the future to study various disorders such as compulsive behaviors, which are present in the Aplysia, and probably have the same origins as in humans, given that many fundamental processes (such as memory studied by Eric Kandel who he demonstrated that human and Aplysia memories works with the same mechanism) are similar between the two species.

With parts of SA in a malaria endemic area, a preventative way of fighting malaria is with the use of pesticides such as 1,1,1–Trichloro–2,2–bis(pclorophenyl) ethane, also known as DDT. DDT is listed under the persistent organic pollutants (POPs) and considered an endocrine disruptive compound (EDC) under the Stockholm Convention. SA registered an exemption to use DDT as means to fight malaria. DDT and its isomers are, however, known EDCs. Combined with their ability to persist in the environment while not being target specific motivates further studies into possible detrimental effects. The present study aimed to establish if ED was present by comparing the male reproductive organs from snails from an area currently sprayed with DDT (for malaria control) to an area not sprayed with DDT in the Limpopo Province. A possible endpoint (the penis sheath/preputium length ratio or PSPLR) was identified for the freshwater snail Bulinus tropicus. B. tropicus and sediment samples were collected from DDT–sprayed and nonsprayed areas located close together. The snails were dissected and various morphometric parameters measured. Sediments from the sites where the snails were collected were analysed for DDT using GC–MS. Statistical analysis showed significant differences in PSPLR (and therefore possible ED) between snails from the two areas. The difference in PSPLR values was mainly due to a relatively shorter preputium for the snails from the DDT–sprayed area. Even though the sediment samples showed that DDT was present in most of the DDT–sprayed sites and not in the non–DDT sprayed sites, causality of the possible ED could not be established from this field study. This study indicated the possibility of using the PSPLR as endpoint for ED. Recommendations are made for further development of the PSPLR and B. tropicus as biological indicators for endocrine disruption, but causality must first be established. / Thesis (M.Sc (Environmental Science))--North-West University, Potchefstroom Campus, 2011.

With parts of SA in a malaria endemic area, a preventative way of fighting malaria is with the use of pesticides such as 1,1,1–Trichloro–2,2–bis(pclorophenyl) ethane, also known as DDT. DDT is listed under the persistent organic pollutants (POPs) and considered an endocrine disruptive compound (EDC) under the Stockholm Convention. SA registered an exemption to use DDT as means to fight malaria. DDT and its isomers are, however, known EDCs. Combined with their ability to persist in the environment while not being target specific motivates further studies into possible detrimental effects. The present study aimed to establish if ED was present by comparing the male reproductive organs from snails from an area currently sprayed with DDT (for malaria control) to an area not sprayed with DDT in the Limpopo Province. A possible endpoint (the penis sheath/preputium length ratio or PSPLR) was identified for the freshwater snail Bulinus tropicus. B. tropicus and sediment samples were collected from DDT–sprayed and nonsprayed areas located close together. The snails were dissected and various morphometric parameters measured. Sediments from the sites where the snails were collected were analysed for DDT using GC–MS. Statistical analysis showed significant differences in PSPLR (and therefore possible ED) between snails from the two areas. The difference in PSPLR values was mainly due to a relatively shorter preputium for the snails from the DDT–sprayed area. Even though the sediment samples showed that DDT was present in most of the DDT–sprayed sites and not in the non–DDT sprayed sites, causality of the possible ED could not be established from this field study. This study indicated the possibility of using the PSPLR as endpoint for ED. Recommendations are made for further development of the PSPLR and B. tropicus as biological indicators for endocrine disruption, but causality must first be established. / Thesis (M.Sc (Environmental Science))--North-West University, Potchefstroom Campus, 2011.