Methamphetamine (MA) abuse is a fast growing drug problem, and is the second most widely abused drug world-wide. MA abuse has been linked to the development of symptoms indistinguishable from schizophrenia, referred to as MA psychosis. MA abusing individuals, who most often comprise adolescents and young adults, are 11 times more likely than the general population to develop psychosis. Of further concern is that in utero exposure to MA is also a growing problem, with more women addicts choosing MA as their primary drug. This has significant implications for the neurodevelopment of the child, with subsequent behavioural deficits later in life. Epidemiological studies suggests that in utero or early life MA exposure places a vulnerable individual at greater risk for developing schizophrenia, although this has never been formerly studied either at clinical or pre-clinical level. Animal models of early life adversity, such as post-weaning social isolation rearing (SIR), can assist in understanding the underlying mechanisms in MA abuse and vulnerability to develop MA psychosis. The aim of the current study was to investigate the long term effects of either prenatal (in utero) or early postnatal administration of MA on the development of schizophrenia-like behavioural and neurochemical abnormalities later in life. In the in utero study, pregnant female Wistar rats received either saline (Sal) or MA 5 mg/kg/day for 16 days by subcutaneous (s.c.) injection , starting on prenatal day 13 (PreND-13) up to postnatal day 2 (PostND02). Male offspring were selected for the study. On PostND 21, the animals were weaned and reared under group or isolation reared conditions for 8 weeks. In the early postnatal study, adult male Wistar rats were divided into group reared and SIR conditions from PostND21. Either group received an escalating dose of MA twice a day (0.2 mg/kg – 6 mg/kg s.c.) or Sal for 16 days, from PostND35 to PostND50. Both in utero and early postnatal groups were then subjected to various behavioural tests on PostND78, including assessment of social interaction (SI) and prepulse inhibition (PPI) of acoustic startle. Following behavioural testing, rats were sacrificed and brains snap frozen for later analysis of cortico-striatal monoamine concentrations, superoxide dismutase activity and lipid peroxidation. In the prenatally exposed group no differences in %PPI was observed, although group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including approaching, time together and anogenital sniffing. SIR animals receiving Sal or MA also showed a decrease in rearing. Regarding self-directed behaviours, group reared animals receiving MA and SIR animals receiving Sal or MA showed an increase in self-grooming. Although some disturbances in regional brain monoamines were observed in the frontal cortex and striatum across the groups, this did not reach significance. A significant increase in malondialdehyde was observed in the striatum in group reared animals receiving MA as well as SIR animals receiving Sal or MA, indicating cell damage, possibly of redox origin. In the early postnatal study, %PPI was significantly reduced in group reared animals receiving MA as well as in SIR animals receiving Sal or MA. Group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including rearing, approaching, time together and anogenital sniffing. Regarding self-directed behaviours and locomotor activity, self-grooming and squares crossed was significantly increased in group reared animals receiving MA and SIR animals receiving Sal or MA. A significant increase in DA was evident in the frontal cortex of SIR and grouped housed animals receiving MA. DA in the MA + SIR combination was elevated but not significantly so. None of the treatments affected striatal monoamine levels. In the group reared animals receiving MA as well as the SIR animals receiving Sal or MA, a significant decrease in SOD activity was observed in the frontal cortex, indicating the presence of oxidative stress in this brain region. None of the parameters indicated an additive effect in MA + SIR treated animals. In conclusion, prenatal exposure to MA led to some evidence of late-life behavioural and neurochemical abnormalities akin to schizophrenia, confirming its penchant for psychotogenic effects. However, chronic postnatal MA exposure was more emphatic, being as effective as SIR, a neurodevelopmental model of schizophrenia, in inducing deficits in the above-mentioned behavioural and neurochemical parameters. Thus, early adolescent abuse of MA is a significant risk factor for the later development of schizophrenia or psychosis. However, the risk appeared not to be exacerbated in a population at risk, i.e. in SIR animals. / Thesis (MSc (Pharmacology))--North-West University, Potchefstroom Campus, 2013.

Methamphetamine (MA) abuse is a fast growing drug problem, and is the second most widely abused drug world-wide. MA abuse has been linked to the development of symptoms indistinguishable from schizophrenia, referred to as MA psychosis. MA abusing individuals, who most often comprise adolescents and young adults, are 11 times more likely than the general population to develop psychosis. Of further concern is that in utero exposure to MA is also a growing problem, with more women addicts choosing MA as their primary drug. This has significant implications for the neurodevelopment of the child, with subsequent behavioural deficits later in life. Epidemiological studies suggests that in utero or early life MA exposure places a vulnerable individual at greater risk for developing schizophrenia, although this has never been formerly studied either at clinical or pre-clinical level. Animal models of early life adversity, such as post-weaning social isolation rearing (SIR), can assist in understanding the underlying mechanisms in MA abuse and vulnerability to develop MA psychosis. The aim of the current study was to investigate the long term effects of either prenatal (in utero) or early postnatal administration of MA on the development of schizophrenia-like behavioural and neurochemical abnormalities later in life. In the in utero study, pregnant female Wistar rats received either saline (Sal) or MA 5 mg/kg/day for 16 days by subcutaneous (s.c.) injection , starting on prenatal day 13 (PreND-13) up to postnatal day 2 (PostND02). Male offspring were selected for the study. On PostND 21, the animals were weaned and reared under group or isolation reared conditions for 8 weeks. In the early postnatal study, adult male Wistar rats were divided into group reared and SIR conditions from PostND21. Either group received an escalating dose of MA twice a day (0.2 mg/kg – 6 mg/kg s.c.) or Sal for 16 days, from PostND35 to PostND50. Both in utero and early postnatal groups were then subjected to various behavioural tests on PostND78, including assessment of social interaction (SI) and prepulse inhibition (PPI) of acoustic startle. Following behavioural testing, rats were sacrificed and brains snap frozen for later analysis of cortico-striatal monoamine concentrations, superoxide dismutase activity and lipid peroxidation. In the prenatally exposed group no differences in %PPI was observed, although group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including approaching, time together and anogenital sniffing. SIR animals receiving Sal or MA also showed a decrease in rearing. Regarding self-directed behaviours, group reared animals receiving MA and SIR animals receiving Sal or MA showed an increase in self-grooming. Although some disturbances in regional brain monoamines were observed in the frontal cortex and striatum across the groups, this did not reach significance. A significant increase in malondialdehyde was observed in the striatum in group reared animals receiving MA as well as SIR animals receiving Sal or MA, indicating cell damage, possibly of redox origin. In the early postnatal study, %PPI was significantly reduced in group reared animals receiving MA as well as in SIR animals receiving Sal or MA. Group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including rearing, approaching, time together and anogenital sniffing. Regarding self-directed behaviours and locomotor activity, self-grooming and squares crossed was significantly increased in group reared animals receiving MA and SIR animals receiving Sal or MA. A significant increase in DA was evident in the frontal cortex of SIR and grouped housed animals receiving MA. DA in the MA + SIR combination was elevated but not significantly so. None of the treatments affected striatal monoamine levels. In the group reared animals receiving MA as well as the SIR animals receiving Sal or MA, a significant decrease in SOD activity was observed in the frontal cortex, indicating the presence of oxidative stress in this brain region. None of the parameters indicated an additive effect in MA + SIR treated animals. In conclusion, prenatal exposure to MA led to some evidence of late-life behavioural and neurochemical abnormalities akin to schizophrenia, confirming its penchant for psychotogenic effects. However, chronic postnatal MA exposure was more emphatic, being as effective as SIR, a neurodevelopmental model of schizophrenia, in inducing deficits in the above-mentioned behavioural and neurochemical parameters. Thus, early adolescent abuse of MA is a significant risk factor for the later development of schizophrenia or psychosis. However, the risk appeared not to be exacerbated in a population at risk, i.e. in SIR animals. / Thesis (MSc (Pharmacology))--North-West University, Potchefstroom Campus, 2013.

Posttraumatic stress disorder (PTSD) is an anxiety disorder that may result from an exposure to a severely traumatic life-event. It is characterised by a delayed onset of psychological and physical symptoms including re-experiencing the event, avoidance of reminders associated with the trauma, increased autonomic arousal and distinct memory deficits. This disorder is also characterised by a maladaptive hypothalamic-pituitary-adrenal (HPA)-axis response and altered monoamine concentrations in the hippocampus and pre-frontal cortex. The Time Dependent Sensitization (TDS) model is a putative animal model of PTSD that is based on the concept of repeated trauma, using three acute stressors (TS) of intense severity followed by a mild situational reminder (RS) on day 7 subsequent to the acute stressors. The aims of this study were to determine if the Triple Stressor (TS) induces stress and if the situational reminder (RS) is necessary for the maintenance of the stress response over time and whether these two stress responses are qualitatively and quantitively different. This was done to further validate the TDS model and to characterize the development and progression of the stress-related pathology of PTSD. Methods used were High Performance Liquid Chromatography (HPLC) with electrochemical detection (biochemical correlates) for quantifying the monoamines dopamine (DA), noradrenaline (NA) and serotonin (5-HT) concentrations in the hippocampus and pre-frontal cortex (PFC); radio immuno assay (RIA) for the determination of plasma corticosterone concentrations (neuroendocrine parameter) and the use of the Elevated Plus Maze (EPM) to detect anxiety-like behaviour (behavioural analyses). The study was subdivided into an Acute and Re-Stress study (n = 10). In the Acute Study rats were exposed to TS as the only stressor. Group 1 was sacrificed immediately after TS, Group 2 was sacrificed 3 days post TS and Group 3 on day 7 post TS. In the Re-Stress Study both TS and RS were used as stressors. Group 4 was sacrificed immediately after the situational reminder, Group 5 was sacrificed 3 days post RS and Group 6 on day 7 post RS. A group of unstressed rats were used as Control. The results of this study found corticosterone concentrations elevated immediately after the TS (p<0.05). Exposure to the RS resulted in a profound hypocortisolism (p<0.05). These results indicate a possible disturbance in the regulation of the HPA-axis, which manifests as an enhanced negative feed-back upon re-introduction of the stressful situation. Changes in MA concentrations were evident. Although no definite fixed trend is apparent in this study, it is evident that the TDS model does induce monoamine dysregulation. Hippocampal NA. DA and 5-HT concentrations were noted to be elevated on day 7 post TS (p<0.05). On day 7 post RS only hippocampal 5HT was decreased significantly (p<0.05). Behavioural analyses indicate that stress related anxiety was not sustained after the TS but 7 days after the exposure to the RS rats were most anxious (p<0.05). The results confirm that the TDS model does induce PTSD-like symptoms in rats and that the situational reminder (RS) is necessary for the maintenance of the stress response. This model may be useful in the investigation of future experimental pharmacological interventions in the management of PTSD. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2005.

Connectomics – the study of neural connectivity – is primarily concerned with the mapping and characterisation of wired synaptic links; however, it is well established that long-distance chemical signalling via extrasynaptic volume transmission is also critical to brain function. As these interactions are not visible in the physical structure of the nervous system, current approaches to connectomics are unable to capture them. This work addresses the problem of missing extrasynaptic interactions by demonstrating for the first time that whole-animal volume transmission networks can be mapped from gene expression and ligand-receptor interaction data, and analysed as part of the connectome. Complete networks are presented for the monoamine systems of Caenorhabditis elegans, along with a representative sample of selected neuropeptide systems. A network analysis of the synaptic (wired) and extrasynaptic (wireless) connectomes is presented which reveals complex topological properties, including extrasynaptic rich-club organisation with interconnected hubs distinct from those in the synaptic and gap junction networks, and highly significant multilink motifs pinpointing locations in the network where aminergic and neuropeptide signalling is likely to modulate synaptic activity. Thus, the neuronal connectome can be modelled as a multiplex network with synaptic, gap junction, and neuromodulatory layers representing inter-neuronal interactions with different dynamics and polarity. This represents a prototype for understanding how extrasynaptic signalling can be integrated into connectomics research, and provides a novel dataset for the development of multilayer network algorithms.

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2017-06-08T20:21:50Z No. of bitstreams: 2 Tese - James Oluwagbamigbe Fajemiroye - 2015.pdf: 1734029 bytes, checksum: c1790bce6e7eace236dd1d68cec686a8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-06-09T11:42:36Z (GMT) No. of bitstreams: 2 Tese - James Oluwagbamigbe Fajemiroye - 2015.pdf: 1734029 bytes, checksum: c1790bce6e7eace236dd1d68cec686a8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-06-09T11:42:36Z (GMT). No. of bitstreams: 2 Tese - James Oluwagbamigbe Fajemiroye - 2015.pdf: 1734029 bytes, checksum: c1790bce6e7eace236dd1d68cec686a8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-10-16 / Depression and anxiety are widely acclaimed as psychiatric disorders of global concern. These disorders are among the leading causes of disability worldwide. Unsatisfactory responses of patients to the available pharmacotherapy make the search for new drugs a necessity. Medicinal plants remain important source of new drugs and new chemical entities. The ethnopharmacological knowledge and previous data have revealed calming and anxiolytic like effects of the organic leaf extract of Pimenta pseudocaryophyllus (Gomes) L.R. Landrum. The present study sought to investigate antidepressive like effect of dichloromethane fraction (DF) of the ethanolic leaf extract of Pimenta pseudocaryophyllus as well as anxiolytic and antidepressive like effects of oleanolic acid (OA), (E) methyl isoeugenol (MIE) and possible mechanisms of action that are involved. Animal models like barbiturate-induced sleep, light dark box test (LDB), elevated plusmaze (EPM), open field (OF), wire hanging test, pentylenetetrazol-induced convulsion test, forced swimming test (FST), tail suspension test (TST) were conducted to evaluate behavioural alterations that were elicited by the administrations of vehicle, DF, OA, MIE or reference drugs. Bioassays (ex vivo and in vitro) of monoamine oxidase (MAO) and quantification of hippocampal level of brain derived neurotrophic factor (BDNF) were conducted in an attempt to elucidate possible mechanisms of action. Oral administration of DF 125, 250 or 500 mg/kg (potentiated the hypnotic effect of sodium pentobarbital). In the TST and FST, DF 125 or 250 mg/kg induced antidepressant-like response. The data obtained in the OF suggest sedative effect of DF at 500 mg/kg. Pretreatment (i.p) with pchlorophenylalanine methyl ester (PCPA) 100 mg/kg (serotonin depletor) or 𝛼-methyl-ptyrosine (AMPT) 100 mg/kg (catecholamine depletor) blocked anti-immobility effect of DF viii in the FST. The enzymatic activity of MAO remained unaltered by DF. Oral administration of OA (5-20 mg/kg) increased the duration of barbiturate - induced sleep and demonstrated anxiolytic like effect in both LDB and EPM. In the FST and TST, OA 5-20 mg/kg elicited antidepressant like effect without altering locomotion activity of the animals. The antidepressant like effect of OA was attenuated by NAN-190 (non-selective antagonist of 5-HT1A), AMPT, PCPA, WAY and PRAZ. Chronic administration of OA increased hippocampal level of BDNF. Oral administration of MIE 250 or 500 mg/kg potentiated hypnotic effect of sodium pentobarbital without protecting mice against PTZ - induced convulsion. The parameters evaluated in the LDB, EPM and OF demonstrated anxiolytic like property of MIE. This effect was blocked by WAY (selective antagonist of 5-HT1A) pretreatment. MIE 125 or 250 mg/kg showed antidepressant like effect in the FST. Locomotion activity of the animal in the OF remained unaltered by MIE administration at 125 or 250 mg/kg. Pretreatment of mice with PCPA attenuated antidepressant like property of MIE. In conclusion, our findings demonstrated anxiolytic and/or antidepressant like effects of dichloromethane fraction, oleanolic acid and (E) methyl isoeugenol, thereby suggesting the involvement of monoaminergic pathway. / Ansiedade e depressão são transtornos psiquiátricos de interesse global. Estes transtornos estão entre as principais causas da incapacidade laboral das pessoas. Apesar de uma gama de farmacoterapias disponíveis, os resultados clínicos mostram que os fármacos não produziram efeitos terapêuticos desejados e se faz necessário a busca de novos fármacos. As plantas medicinais continuam sendo uma das fontes mais importantes para a descoberta de novos fármacos e entidades químicas. Estudos anteriores mostraram efeito calmante e ansiolítico da fração orgânica do extrato das folhas de Pimenta pseudocaryophyllus (Gomes) L. R. Landrum (Myrtaceae). O presente estudo buscou investigar a atividade tipo antidepressiva da fração diclorometano (FD) do extrato etanólico das folhas desta espécie, bem como antidepressiva e ansiolítica do ácido oleanólico (AO), (E)-metilisoeugenol (MIE) e os possíveis mecanismos de ações envolvidos. Modelos experimentais como o sono induzido por barbitúricos, caixa claro escuro (CCE), labirinto em cruz elevado (LCE), campo aberto (CA), teste de arame, teste de convulsão induzida por pentilenotetrazol, teste de natação forçada (TNF) e teste de suspensão pela cauda (TSC) foram realizados para avaliar alterações comportamentais induzidas pela administração do veículo, FD, AO, MIE ou fármacos de referência. Na tentativa de elucidar os possíveis mecanismos de ação, foram realizados bioensaios (ex vivo e in vitro) da monoamina oxidase (MAO) e do fator neurotrófico derivado do cérebro (BDNF do hipocampo). A administração oral da FD 125, 250 ou 500 mg/kg potencializou o efeito hipnótico de pentobarbital sódico. No TNF e TSC, a FD 125 ou 250 mg/kg induziu efeito tipo antidepressivo. Os dados obtidos no campo aberto sugerem efeito sedativo da fração vi diclorometano na dose de 500 mg/kg. O pré-tratamento (i.p) com p - clorofenilalanina metil éster (PCPA) 100 mg/kg (depletor de serotonina) ou α - metil - p - tirosina (AMPT) 100 mg/kg (depletor de catecolamina) bloqueou o efeito tipo antidepressivo da FD no TNF. O bioensaio da atividade enzimática mostrou que a FD não alterou a atividade da MAO. A administração oral do AO (5-20 mg/kg) aumentou a duração do sono induzido por pentobarbital sódico e demonstrou efeito tipo ansiolítico no CCE e LCE. O AO 5-20 mg/kg demonstrou efeito tipo antidepressivo no TNF e TSC sem alterar a atividade locomotora dos animais. O efeito tipo antidepressivo do AO foi atenuado por prétratamento com NAN-190 (antagonista não-seletivo do receptor 5-HT1A), AMPT, PCPA e PRAZ-prazosin (antagonista do receptor α1 adrenérgico). A administração crônica do AO aumentou o nível de BDNF no hipocampo. A administração oral do MIE 250 ou 500 mg/kg potencializou o efeito hipnótico de pentobarbital sódico sem proteger os animais contra a convulsão induzida por PTZ. Os parâmetros avaliados na CCE e LCE sugerem que MIE têm efeito tipo ansiolítico. Este efeito foi bloqueado pelo pré-tratamento com WAY100635 (antagonista seletivo do receptor 5-HT1A). MIE 125 ou 250 mg/kg apresentou efeito tipo antidepressivo no TNF. Não houve alteração na atividade locomotora dos animais no CA após a administração do MIE 125 ou 250 mg/kg. O prétratamento com PCPA atenuou o efeito tipo antidepressivo do MIE no TNF. Os resultados demonstraram efeito tipo ansiolítico e/ou antidepressivo da fração diclorometano, ácido oleanólico e (E)-metilisoeugenol, sugerindo o envolvimento de vias monoaminérgicas nestes efeitos.

Tesis por compendio / [ES] El sistema de melanocortina es una estructura clave en la regulación de una amplia gama de funciones fisiológicas que incluyen la melanogénesis, la respuesta al estrés y el equilibrio energético, mediante la unión a una familia de receptores acoplados a la proteína G específicos (MC1R-MC5R). La sobreexpresión de agonistas inversos, la proteína de señalización agutí (Asip) y la proteína relacionada con agutí (Agrp) da como resultado un aumento de la ingesta de alimentos, de crecimiento lineal y de peso corporal. Asip regula la polaridad de pigmentación dorsoventral a través del MC1R, y la sobreexpresión induce obesidad en ratones al unirse al Mc4r central. La sobreexpresión de asip1 en el pez cebra transgénico (asip1-Tg) mejora el crecimiento, sin afectar la acumulación lipídica (obesidad), incluso cuando se alimentan bajo regímenes inductores severos. Los peces asip1-Tg no necesitan comer más para crecer más y más rápido, lo que sugiere una mayor eficiencia alimentaria. Además, los peces asip1-Tg criados en alta densidad son capaces de crecer mucho más que los peces de tipo salvaje (WT) criados en baja densidad, aunque los peces asip1-Tg parecen ser más sensibles al estrés por hacinamiento que los peces WT. El análisis transcriptómico comparativo del intestino de asip1-Tg refleja una expresión diferencial de transportadores aminoácidicos, monocarboxilatos, transportadores iónicos y de vitaminas. La sobreexpresión reduce la integridad del epitelio intestinal aumentando su permeabilidad paracelular y potencia el transporte electrogénico de aminoácidos. Así, los peces transgénicos poseen mayor capacidad para la absorción de nutrientes y, por extensión una mejora en la eficiencia alimenticia que podría explicar, en parte, ese crecimiento diferencial bajo tasas de ingesta similares. Esta tesis tuvo también como objetivo investigar si los asip1-Tg mantienen un fenotipo dominante asociado con una mayor tasa de alimentación. Los resultados muestran, por el contrario, un carácter reactivo/subordinado en los asip1-Tg que aboga por una participación del sistema de melanocortinas en la regulación del comportamiento de peces. El perfil subordinado de los animales asip1-Tg, junto con una activación del eje del estrés, sugiere que estos animales pueden mostrar un comportamiento de ansiedad. Los resultados indicaron que los peces asip1-Tg muestran un comportamiento de ansiedad que además relacionado con una severa disminución de los niveles centrales de serotonina (5HT) y dopamina y elevación de su recaptación neuronal y degradación. La administración de un inhibidor de la recaptación de 5HT, recupera el fenotipo comportamental salvaje, mitigando el comportamiento de ansiedad en los peces transgénicos y rescatando los niveles de 5HT. Esta ansiedad podría repercutir en una alteración del comportamiento locomotor de los animales, por ello estudiamos los ritmos circadianos de actividad locomotora. Los resultados muestran que los animales asip1-Tg exhiben una disrupción completa del ritmo de actividad, con una actividad muy elevada, especialmente durante la noche. Esta disrupción es concomitante con una desaparición del ritmo diario de serotonina y melatonina. Además, los resultados muestran una pérdida de ritmos de expresión de genes reloj (per1a y clock1a). La incubación, in vitro, de glándulas pineales con Asip1 produjo una inhibición de la secreción de melatonina replicando los resultados obtenidos in vivo y demostrando un efecto directo de Asip1, sobre la fisiología de la pineal. En esta tesis, se utilizó el pez cebra como modelo para investigar los efectos de la obesidad sobre la ansiedad y la memoria. La obesidad no tuvo ningún efecto sobre la ansiedad, pero produjo una disminución de la memoria a corto plazo, estudiada mediante test de condicionamiento aversivo. Este estudio proporciona, un protocolo fiable para evaluar el efecto de las enfermedades metabólica en la función cognitiva y conductual. / [CA] El sistema de melanocortina és una estructura clau en la regulació d'una ampla gamma de funcions fisiològiques que inclouen la melanogènesi, la resposta a l'estrès i l'equilibri energètic, mitjançant la unió a una família de receptors acoblats a la proteïna G específics (MC1R-MC5R). La sobreexpressió d'agonistes inversos, la proteïna de senyalització agutí (Asip) y la proteïna relacionada con agutí (Agrp) dona com a resultat un augment de la ingesta d'aliments, de creixement lineal i de pes corporal. Asip regula la polaritat de pigmentació dors-ventral a través del MC1R, y la sobreexpressió indueix obesitat en ratolins en unir-se al MC4R central. La sobreexpressió de asip1 en el peix zebra transgènic (asip1-Tg) millora el creixement, sense afectar l'acumulació lipídica (obesitat), inclús quan s'alimenten sota règims inductors severs. Los peces asip1-Tg no necessiten menjar més per a créixer més i més ràpid, lo qual suggereix una major eficiència alimentària. A més a més, els peixos asip1-Tg criats en alta densitat són capaces de créixer molt més que els peixos de tipus salvatge (WT) criats en baixa densitat, malgrat que els peixos asip1-Tg semblen ser més sensibles a l'estrès per amuntegament que els peixos WT. L'anàlisi transcriptòmic comparatiu de l'intestí de asip1-Tg reflecteix una expressió diferencial de transportadors aminoacídics, monocarboxilats, transportadors iònics i de vitamines. La sobreexpressió redueix la integritat de l'epiteli intestinal augmentant la seua permeabilitat paracel·lular i potencia el transport electrogènic d'aminoàcids. Per tant, els peixos transgènics posseeixen major capacitat per l'absorció de nutrients i, per extensió una millora en la eficiència alimentària que podria explicar, en part, eixe creixement diferencial sota taxes d'ingesta similars. Aquesta tesi tingué també com a objectiu investigar si els asip1-Tg mantenien un fenotip dominant associat amb una major taxa d'alimentació. Els resultats mostren, pel contrari, un caràcter reactiu/subordinat en los asip1-Tg que advoca per una participació del sistema de melanocortines en la regulació del comportament de peixos. El perfil subordinat dels animals asip1-Tg, junt amb una activació de l'eix de l'estrès, suggereix que aquests animals poden mostrar un comportament d'ansietat. Els resultats indicaren que els peixos asip1-Tg mostren un comportament d'ansietat relacionat amb una severa disminució dels nivells centrals de serotonina (5HT) i dopamina i elevació de la seua recaptació neuronal i degradació. L'administració de un inhibidor de la recaptació de 5HT recupera el fenotip comportamental salvatge, mitigant el comportament d'ansietat en els peixos transgènics i rescatant els nivells centrals de 5HT. Esta ansietat podria repercutir en una alteració del comportament locomotor dels animals, per la qual cosa vam estudiar els ritmes circadians d'activitat locomotora. Els resultats mostren que els animals asip1-Tg exhibeixen una disrupció completa del ritme d'activitat, amb una activitat molt elevada durant tot el cicle diari, especialment durant la nit. Esta disrupció es concomitant amb una desaparició del ritme diari de serotonina i melatonina. A més a més, els resultats mostren una pèrdua de ritmes de expressió de gens rellotge (per1a y clock1a). La incubació, in vitro, de glàndules pineals con Asip1 va produir una inhibició de la secreció de melatonina replicant els resultats obtinguts in vivo y demostrant un efecte directe de Asip1 sobre la fisiologia de la pineal. En esta tesi, se va utilitzar el peix zebra com a model per investigar els efectes de la obesitat sobre la ansietat i la memoria. L'obesitat no va tindre cap efecte sobre l'ansietat, però va produir una disminució de la memòria a curt termini, estudiada mitjançant tests de condicionament aversiu. Aquest estudi proporciona, un protocol fiable per a avaluar l'efecte de les malalties metabòliques en la funció cognitiva i conductual. / [EN] The melanocortin system plays a key role in the regulation of a wide range of physiological functions including melanogenesis, stress response and energy balance, through binding to a family of specific G protein-coupled receptors (MC1R-MC5R). Overexpression of inverse agonists, agouti-signalling protein (Asip) and agouti-related protein (Agrp) results in increased food intake, linear growth and body weight. Asip regulates dorso-ventral pigmentation polarity through MC1R, and over-expression induces obesity in mice by binding to the central MC4R. Overexpression of asip1 in transgenic zebrafish (asip1-Tg) enhances growth, without affecting lipid accumulation (obesity), even when fed under severe inducing regimens. The asip1-Tg fish do not need to eat more to grow bigger and faster, suggesting increased feed efficiency. In addition, asip1-Tg fish reared at high density are able to grow much larger than wild-type (WT) fish reared at low density, although asip1-Tg fish appear to be more sensitive to overcrowding stress than WT fish. Comparative transcriptomic analysis of asip1-Tg gut reflects differential expression of amino acid, monocarboxylate, ionic and vitamin transporters. Overexpression reduces the integrity of the intestinal epithelium by increasing its paracellular permeability and enhances electrogenic amino acid transport. Thus, transgenic fish possess a greater capacity for nutrient absorption and, by extension, an improvement in feed efficiency that could explain, in part, this differential growth under similar intake rates. This thesis also aimed to investigate whether asip1-Tg maintain a dominant phenotype associated with a higher feeding rate. Experimental results show, on the contrary, a reactive/subordinate character in asip1-Tg which argues for an involvement of the melanocortin system in the regulation of fish behaviour. Improving feeding motivation without promoting aggression in fish, thus avoiding the threat to native populations in case of an escape, makes the inhibition of the melanocortin system, through the overexpression of asip1, a feasible target for the development of genetically modified lines. The subordinate profile of the asip1-Tg animals, together with an activation of the stress axis, suggests that these animals may exhibit anxiety-like behaviour. The results indicated that asip1-Tg fish show a behaviour similar to our concept of anxiety related to a severe decrease in central serotonin (5HT) and dopamine levels as well as the elevation of their neuronal reuptake and degradation. The administration of fluoxetine, a serotonin reuptake inhibitor, recovers the wild-type behavioural phenotype, mitigating anxiety behaviour in transgenic fish and restoring central 5HT levels. This anxiety could have repercussions on the locomotor behaviour of the animals, so we studied circadian rhythms of locomotor activity. The results show that asip1-Tg animals exhibit a complete disruption of the activity rhythm, with very high activity levels throughout the daily cycle, especially during the night. This disruption is concomitant with a disappearance of the daily rhythm of serotonin and melatonin. In addition, the results show a loss of clock gene expression rhythms (per1a and clock1a). Incubation, in vitro, of pineal glands with Asip1 produced an inhibition of melatonin secretion replicating the results obtained in vivo and demonstrating a direct effect of Asip1 on pineal physiology. In this PhD thesis, zebrafish was used as a model to investigate the effects of overfeeding-induced obesity on anxiety-like behaviour and memory. Obesity had no effect on anxiety, but produced a decrease in short-term memory, studied by means of aversive conditioning tests. This study also provides a reliable protocol for assessing the effect of metabolic diseases on cognitive and behavioural function, supporting zebrafish as a model for cognitive and behavioural neuroscience. / Esta tesis ha sido realizada a través del programa de ‘Ayudas para la formación de personal investigador’ (FPI) BES‐2017‐082424 de la Agencia Estatal de Investigación, en Instituto de Acuicultura Torre de la Sal (IATS) del Consejo Superior de Investigaciones Científicas (CSIC) en el Grupo de investigación de Control de la Ingesta en Peces dirigido por José Miguel Cerdá Reverter, director de esta tesis. Los trabajos llevados a cabo en esta tesis han sido financiados por Ministerio de Ciencia, Innovación y Universidades (MICIU) a través de los siguientes proyectos: MELANOCONDUCT: Implicación del sistema de melanocortinas en la regulación de los mecanismos temporales y conductuales de peces AGL2016-74857-C3-3-R; Cronopeces: Red temática de cronobiología de peces y sus aplicaciones en acuicultura RED2018-102487-T; MacForFish: Nuevos aspectos homeostáticos y comportamentales de la regulación de la ingesta en peces PID2019-103969RB-C33; FISHTASTE: Implicación de los mecanismos sensoriales del gusto en la regulación de la ingesta de peces - Involvement of taste sensing mechanisms in the regulation of feed intake of fish PID2022-136288OB-C33 / Godino Gimeno, A. (2024). Involvement of the Melanocortin System in the Regulation of Circadian and Behavioural Mechanisms in Zebrafish [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/203148 / Compendio

Methamphetamine (METH) abuse has become a fast growing drug problem that has developed into a global epidemic. In fact, METH is one of the most commonly abused substances with an estimated 35 million abusers worldwide and is said to be the second most popular illicit drug. The Western Province of South Africa has seen a dramatic increase in drug abuse in recent years where METH is the primary or secondary drug of abuse. Interestingly, more than 50% of these individuals are under the age of 20 years. The longer duration of euphoric effects of METH has attracted many users away from cocaine in favour of METH. In addition to the rapid euphoric effect of METH, the direct short-term effects include arousal, reduced fatigue, an increase in blood pressure, reduced appetite as well as sustained attention. Chronic METH abuse may result in debilitating and long-lasting effects that includes mood disorders such as depression. Studies suggest a strong relationship between exposure to adverse environmental factors early in life and the later development of a neuropsychiatric disorder, such as depression. However, these severe consequences do not seem to invoke cessation of the drug. The euphoric and addictive properties of METH causes users to abuse the drug with an increase in frequency and dose, even though it might not have been their original intention. The primary objective of this study was to investigate the effect of early-life administration of METH to stress-sensitive (Flinders Sensitive Line - FSL) and control (Flinders Resistant Line - FRL) rats on depressive-like behaviour and regional brain monoamine levels later in life. The study implemented a sixteen-day period for administration of METH or a vehicle control from postnatal day 19 (PnD19) to postnatal day 34 (PnD34). The latter developmental stage corresponds to pre-adolescence in the rat when neurological development are similar to that seen in human adolescents, and represents the stage when drug abuse is most common in humans. Chronic dosing of METH and saline was performed twice daily at 09:00 and at 15:00. The animals received a sub-cutaneous (SC) escalating dose regimen of METH during the 16 day period (mimicking binging behaviour in humans), with every dose escalating in increments of 0.2 mg/kg from 0.2 mg/kg to 6.0 mg/kg. The study then investigated whether early-life administration of METH would cause depressive-like behaviours directly after the injection period (immediate drug effects before withdrawal on PnD35) or later in life (after the withdrawal period in early adulthood on PnD60). The behavioural effects were assessed in a battery of tests and thereafter the rats were sacrificed and the frontal cortex removed and snap frozen for later analyses of altered neurochemistry. The study demonstrated that chronic METH treatment during pre-adolescence induces significant behavioural changes related to depression in humans directly after the injection period (PnD35) and later in life (PnD60). The animals displayed antidepressant-like behaviour in the forced swim test (FST) before withdrawal, yet a depressogenic effect was observed 25 days post-withdrawal. This effect also seems to be additive to the congenital depressive-like phenotype of FSL rats, suggesting a role for genetic susceptibility. This observation would be in line with the two-hit hypothesis of depression, suggesting that the manifestation of depression will result when a genetic predisposition is followed by an environmental stressor (i.e. METH) later in life. The data suggests a working hypothesis that individuals that already have a predisposition to depression may be more susceptible to developing depression when abusing METH. The fact that the FSL control rats were more immobile than FRL control rats also confirmed the face validity of the FSL genetic rat model of depression. Locomotor activity assessment indicated that METH treatment decreased locomotor activity in FSL and FRL rats compared to their vehicle controls on PnD35 but not on PnD60. It is important to note that the effects observed in locomotor activity could not have contributed to the immobility observed in the FST, confirming that the immobility in the FST indeed reflects psychomotor and not locomotor effects. The study also demonstrated that METH significantly lowers social interaction behaviour in both FRL and FSL rats, both immediately following drug treatment (PnD35) and after withdrawal (PnD60). It is therefore clear that this effect of METH is long-lasting, putatively related to neurodevelopmental effects. In addition, the rats investigated the familiar object for a greater amount of time in the novel object recognition test (nORT) on PnD35 and PnD60 and may be the result of loss of recognition memory for the familiar object. This data confirms that METH results in cognitive memory deficits probably due to sustained adverse neurodevelopmental effects. Neurochemical analyses of the frontal cortex indicated decreased serotonin (5-HT) and norepinephrine (NE) levels on PnD35. METH is widely recognised for its pro-inflammatory effects, while the reduced 5-HT levels observed may have been the result of an increase in circulating pro-inflammatory cytokines. Neurochemical analyses provided thought-provoking data concerning the role of the permissive hypotheses of depression, indicating that dopamine (DA) is most likely not responsible for the behavioural effects observed, at least under the current study conditions, whereas 5-HT is decidedly more involved than expected. The data also suggest that depletion in NE plays a role in the development of depressive-like behaviours following METH exposure. Based on these findings, we propose that disturbances in 5-HT and NE are a crucial mechanism in how METH abuse may precipitate or worsen depressive-like symptoms in individuals who abuse METH. It should be noted that this study does not discard the role of DA in the development of depression after METH exposure, although under the current study conditions it appears that DA does not play a central role. The current study demonstrated that pre-adolescent exposure to METH can reproduce most of the behavioural changes seen in depressed individuals, and that these behavioural data can be used to identify causal neurochemical factors. Environmental stressors such as METH abuse should be regarded as an additional diagnostic criterion and is relevant to an accumulative risk factor hypothesis. Furthermore, although further study is required, the data suggests that early-life exposure to METH may predispose an individual to mood disorders and behavioural abnormalities later in life. / MSc (Pharmacology), North-West University, Potchefstroom Campus, 2015

Methamphetamine (METH) abuse has become a fast growing drug problem that has developed into a global epidemic. In fact, METH is one of the most commonly abused substances with an estimated 35 million abusers worldwide and is said to be the second most popular illicit drug. The Western Province of South Africa has seen a dramatic increase in drug abuse in recent years where METH is the primary or secondary drug of abuse. Interestingly, more than 50% of these individuals are under the age of 20 years. The longer duration of euphoric effects of METH has attracted many users away from cocaine in favour of METH. In addition to the rapid euphoric effect of METH, the direct short-term effects include arousal, reduced fatigue, an increase in blood pressure, reduced appetite as well as sustained attention. Chronic METH abuse may result in debilitating and long-lasting effects that includes mood disorders such as depression. Studies suggest a strong relationship between exposure to adverse environmental factors early in life and the later development of a neuropsychiatric disorder, such as depression. However, these severe consequences do not seem to invoke cessation of the drug. The euphoric and addictive properties of METH causes users to abuse the drug with an increase in frequency and dose, even though it might not have been their original intention. The primary objective of this study was to investigate the effect of early-life administration of METH to stress-sensitive (Flinders Sensitive Line - FSL) and control (Flinders Resistant Line - FRL) rats on depressive-like behaviour and regional brain monoamine levels later in life. The study implemented a sixteen-day period for administration of METH or a vehicle control from postnatal day 19 (PnD19) to postnatal day 34 (PnD34). The latter developmental stage corresponds to pre-adolescence in the rat when neurological development are similar to that seen in human adolescents, and represents the stage when drug abuse is most common in humans. Chronic dosing of METH and saline was performed twice daily at 09:00 and at 15:00. The animals received a sub-cutaneous (SC) escalating dose regimen of METH during the 16 day period (mimicking binging behaviour in humans), with every dose escalating in increments of 0.2 mg/kg from 0.2 mg/kg to 6.0 mg/kg. The study then investigated whether early-life administration of METH would cause depressive-like behaviours directly after the injection period (immediate drug effects before withdrawal on PnD35) or later in life (after the withdrawal period in early adulthood on PnD60). The behavioural effects were assessed in a battery of tests and thereafter the rats were sacrificed and the frontal cortex removed and snap frozen for later analyses of altered neurochemistry. The study demonstrated that chronic METH treatment during pre-adolescence induces significant behavioural changes related to depression in humans directly after the injection period (PnD35) and later in life (PnD60). The animals displayed antidepressant-like behaviour in the forced swim test (FST) before withdrawal, yet a depressogenic effect was observed 25 days post-withdrawal. This effect also seems to be additive to the congenital depressive-like phenotype of FSL rats, suggesting a role for genetic susceptibility. This observation would be in line with the two-hit hypothesis of depression, suggesting that the manifestation of depression will result when a genetic predisposition is followed by an environmental stressor (i.e. METH) later in life. The data suggests a working hypothesis that individuals that already have a predisposition to depression may be more susceptible to developing depression when abusing METH. The fact that the FSL control rats were more immobile than FRL control rats also confirmed the face validity of the FSL genetic rat model of depression. Locomotor activity assessment indicated that METH treatment decreased locomotor activity in FSL and FRL rats compared to their vehicle controls on PnD35 but not on PnD60. It is important to note that the effects observed in locomotor activity could not have contributed to the immobility observed in the FST, confirming that the immobility in the FST indeed reflects psychomotor and not locomotor effects. The study also demonstrated that METH significantly lowers social interaction behaviour in both FRL and FSL rats, both immediately following drug treatment (PnD35) and after withdrawal (PnD60). It is therefore clear that this effect of METH is long-lasting, putatively related to neurodevelopmental effects. In addition, the rats investigated the familiar object for a greater amount of time in the novel object recognition test (nORT) on PnD35 and PnD60 and may be the result of loss of recognition memory for the familiar object. This data confirms that METH results in cognitive memory deficits probably due to sustained adverse neurodevelopmental effects. Neurochemical analyses of the frontal cortex indicated decreased serotonin (5-HT) and norepinephrine (NE) levels on PnD35. METH is widely recognised for its pro-inflammatory effects, while the reduced 5-HT levels observed may have been the result of an increase in circulating pro-inflammatory cytokines. Neurochemical analyses provided thought-provoking data concerning the role of the permissive hypotheses of depression, indicating that dopamine (DA) is most likely not responsible for the behavioural effects observed, at least under the current study conditions, whereas 5-HT is decidedly more involved than expected. The data also suggest that depletion in NE plays a role in the development of depressive-like behaviours following METH exposure. Based on these findings, we propose that disturbances in 5-HT and NE are a crucial mechanism in how METH abuse may precipitate or worsen depressive-like symptoms in individuals who abuse METH. It should be noted that this study does not discard the role of DA in the development of depression after METH exposure, although under the current study conditions it appears that DA does not play a central role. The current study demonstrated that pre-adolescent exposure to METH can reproduce most of the behavioural changes seen in depressed individuals, and that these behavioural data can be used to identify causal neurochemical factors. Environmental stressors such as METH abuse should be regarded as an additional diagnostic criterion and is relevant to an accumulative risk factor hypothesis. Furthermore, although further study is required, the data suggests that early-life exposure to METH may predispose an individual to mood disorders and behavioural abnormalities later in life. / MSc (Pharmacology), North-West University, Potchefstroom Campus, 2015

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal