The objective of the present study was to examine the effects of prenatal exposure to alcohol on sensorimotor coordination, emotionality, learning and memory in young adult Wistar rats. Most of the recent reports concerning behavioral effects of fetal alcohol exposure refer to the juvenile period of life and very few studies investigated different aspects of behavior simultaneously in the same subjects. In the current study, alcohol was delivered to the pregnant dams by intragastric infusions, throughout gestation days (GD) 7-20, at the dose of 6g /kg maternal body weight /day. This dose resulted in relatively high peak blood alcohol concentration (340 mg/dl) as assessed on GD 20. A pair-fed isocaloric and untreated control groups were included. Prenatal alcohol administration retarded dams&rsquo / weight gain significantly, and had an adverse effect on pups&rsquo / weight at birth but not in adulthood. No between-group differences were observed in the litter size and in the pups&rsquo / mortality. The adult brain weight was neither affected. Pups were subjected to a series of behavioural tests as young adults (at 2.5 months of age). In adulthood, rats prenatally treated with alcohol were not impaired in sensorimotor coordination and/or did not show muscle weakness as assessed by rotarod/accelerod tests. Their behavior in the open field and plus maze suggested alcohol-induced increase in iv anxiety level and some decrease in behavioral flexibility, but hyperactivity was not observed. In cognitive tasks, alcohol treated rats showed slightly slower rate of initial place learning in the water maze. However, memory retention tested after 1 and 10-day delay, reversal learning, rate of extinction of place preference, as well as working memory capacity appeared to be the same in alcohol exposed and control rats. The possible reasons of this negative result are discussed.

The increasing level of the UV-B irradiation due to depleting of the ozone layer is considered to be one of the causes of global amphibian declines. The UV-B radiation causes damage to the DNA in the nuclei of the skin cells and their eventual death. In many amphibian species there was found a negative influence of the UV-B radiation on the hatching success of tadpoles and their subsequent viability. It has been assumed that tadpoles are able to actively avoid places with the higher exposure of the UV-B radiation thanks to their locomotion. In this experiment tadpoles could have chosen between a zone with the UV radiation (UV-B and UV-A) and without the UV radiation. The tadpoles have not been exposed to the UV radiation before the measurement started. The location of the tadpoles during the measurement was recorded by CCD camera. A computer program EthoVision then evaluated the time that these tadpoles spent in each of the zones and the total path length these tadpoles has swum in the zones. The measurements were performed for two groups of tadpoles differing in age to determine whether the level of development may affect the tadpoles' preference to the UV radiation. The tadpoles in both groups spent more time in the zone without the UV radiation. The tadpoles in the more advanced development phase have spent 7 times more time in average in the zone without the UV radiation than in the zone with it. The tadpoles from the second group have spent in average 2.5 times more time in the zone without the UV radiation than in the zone with the UV radiation. The total swimming path length of advanced tadpoles was almost 2.5 times longer in the zone without the UV radiation than in the zone with the UV radiation. Conversely, the total path length of younger tadpoles did not significantly differ between the zones. The total time spent in the zones with / without the UV radiation did not differ between the groups of tadpoles. Also the total path length did not differ between the groups in the zone with the UV-B radiation. However, the total path length of tadpoles had differed between groups in the zone without the UV radiation.

The hypothesis that experimental manipulations increasing life span correlate to reduced physical activity and reduced molecular oxidative damage was tested. We used three longevity phenotypes of males and females of Pyrrhocoris apterus, diapause insects, reproductive insects and insect with ablation of the corpus allatum, an endocrine gland producing juvenile hormone. Protein carbonyl content in thoracic muscles was used as an index of molecular oxidative modification.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / O Transtorno do Déficit de Atenção e Hiperatividade (TDAH) é uma desordem neurocomportamental caracterizada por graus variados de desatenção, atividade motora excessiva e impulsividade. Sua etiologia não é completamente conhecida, porém, um grande número de evidências sugere que, além de fatores de risco genéticos, a exposição gestacional ao etanol e a altas doses de cafeína contribuem para a manifestação deste transtorno. O etanol, presente na composição de bebidas alcoólicas, e a cafeína, presente na composição de diversas bebidas (refrigerantes, chás, café, e energéticos), alimentos derivados do cacau e medicamentos estão entre as substâncias neuroativas mais consumidas no mundo. Apesar das evidências epidemiológicas do uso combinado de cafeína e etanol por gestantes, as interações entre estas substâncias têm recebido pouca atenção em estudos experimentais. Este fato é particularmente importante visto que alguns estudos apontam que cafeína é capaz de ampliar alguns aspectos importantes da ação de outras substâncias com potencial neurotóxico. Nesse estudo avaliamos os efeitos da co-exposição à cafeína e ao etanol durante o desenvolvimento na atividade locomotora em camundongos adolescentes. Para tanto, camundongos Suíços foram expostos do primeiro dia gestacional até o vigésimo primeiro dia pós-natal (PN21), a solução de cafeína 0,1g/L (Grupo CAF1, 10 ninhadas), a solução de cafeína 0,3g/L (Grupo CAF3, 10 ninhadas) ou tiveram acesso à água potável (Grupo CAF0, 10 ninhadas). Em dias alternados de PN2 a PN8, os animais de cada ninhada receberam uma injeção intraperitoneal de 0,25 l/g de etanol (grupo ETOH25), 0,5 l/g de etanol (grupo ETOH50) ou de solução salina (grupo ETOH0). Em PN30, a atividade locomotora foi avaliada por 15 minutos no teste de Campo Aberto. A análise dos níveis de etanol sérico, realizada em uma amostra independente de animais em PN8, indicou que, para as duas doses de etanol, a alcoolemia dos animais do grupo CAF3 foi significativamente maior do que as dos grupos CAF0 e CAF1, que não diferiram entre si. No teste de campo aberto, apenas os animais expostos ao etanol apresentaram aumento da atividade locomotora. Tanto o grupo ETOH25 quanto o grupo ETOH50 tiveram a atividade maior que o grupo ETOH0. A exposição à cafeína, por si só, não afetou a atividade locomotora dos animais nem potencializou os efeitos do etanol. No grupo CAF3, a atividade locomotora não foi afetada pela exposição ao etanol. Nossos dados confirmam o papel da exposição precoce ao etanol na manifestação da hiperatividade locomotora. Além disso, a exposição à cafeína durante o desenvolvimento pode exercer um papel protetor para a manifestação da hiperatividade locomotora induzida pela exposição precoce ao etanol. / The attention deficit hyperactivity disorder (ADHD) is a neurobehavioral disorder characterized by varying degrees of inattention, excessive motor activity and impulsivity. This etiology is not fully known, but considerable evidence suggests that, in addition to genetic factors, exposure to ethanol and high doses of caffeine contribute to the manifestation of the disorder. Ethanol, which is present in all alcoholic beverages, and caffeine, which is present in many beverages (sodas, teas, coffee and energetic drinks), food derived from cacao and medications, are two of the most consumed neuroactive drugs in the world. In spite of the epidemiological evidence of the combined use of caffeine and ethanol by pregnant women, the interactions between these two drugs have received little attention in experimental studies. This fact is particularly important since some studies have shown that caffeine is capable of amplifying important aspects of the action of other substances that have neurotoxic potential. In the current study we analyzed the effects of co-exposure to caffeine and ethanol during development on the locomotor activity of pre-pubertal mice. In this sense, Swiss mice were exposed from the first gestational day to the 21st postnatal (PN21) day to one of three solutions: 1) a solution containing caffeine 0.1 g/L (group CAF1, 10 litters); 2) a solution containing caffeine 0.3g/L (group CAF3, 10 litters); 3) filtered tap water (group CAF0, 10 litters). Every other day, from PN2 to PN8, animals in each litter were i.p injected with one of the following solutions: 1) 0.25 l/g ethanol (group ETOH25); 2) 0.5 l/g ethanol (group ETOH50); 3) saline solution (group ETOH0). On PN30, locomotor activity was analyzed for 15 min in the open field. The analysis of the ethanol serum levels, which was carried out in an independent sample of animals at PN8, indicated that levels were significantly higher in the CAF3 group when compared to the other two groups, which did not differ between them. In the open field, the ethanol exposure caused a dose-dependent increase in the locomotor activity. The locomotor activity of the group ETOH0 was lower than that presented by the ETOH25 and the ETOH50 groups. The caffeine exposure was not capable of affecting the locomotor activity. In the group CAF3, the locomotor activity was not affected by the ethanol exposure. Our data corroborate previous studies in that it shows that precocious ethanol exposure results in locomotor hyperactivity. Furthermore, it is possible that caffeine exposure during development have a protective effect on deleterious effects of ethanol.

Made available in DSpace on 2014-12-17T15:37:19Z (GMT). No. of bitstreams: 1 ManaraBBCD_DISSERT.pdf: 1554643 bytes, checksum: e4b820c02db19b42acca3889a1b08851 (MD5) Previous issue date: 2014-04-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Neuropeptide S (NPS) is an endogenous 20-aminoacid peptide which binds a G protein-coupled receptor named NPSR. This peptidergic system is involved in the modulation of several biological functions, such as locomotion, anxiety, nociception, food intake and motivational behaviors. Studies have shown the participation of NPSR receptors in mediating the hyperlocomotor effects of NPS. A growing body of evidence suggests the participation of adenosinergic, dopaminergic and CRF systems on the hyperlocomotor effects of NPS. Considering that little is known about the role of dopaminergic system in mediating NPS-induced hyperlocomotion, the present study aims to investigate the locomotor actions of intracerebroventricular (icv) NPS in mice pretreated with α-metil-p-tirosine (AMPT, inhibitor of dopamine synthesis), reserpine (inhibitor of dopamine vesicle storage) or sulpiride (D2 receptor antagonist) in the open field test. A distinct group of animals received the same pretreatments described above (AMPT, reserpine or sulpiride) and the hyperlocomotor effects of methylphenidate (dopamine reuptake inhibitor) were investigated in the open field. NPS and methylphenidate increased the mouse locomotor activity. AMPT per se did not change the locomotion of the animals, but it partially reduced the hyperlocomotion of methylphenidate. The pretreatment with AMPT did not affect the psychostimulant effects of NPS. Both reserpine and sulpiride inhibited the stimulatory actions of NPS and methylphenidate. These findings show that the hyperlocomotor effects of methylphenidate, but not NPS, were affected by the pretreatment with AMPT. Furthermore, methylphenidate- and NPS-induced hyperlocomotion was impaired by reserpine and sulpiride pretreatments. Together, data suggests that NPS can increase locomotion even when the synthesis of catecholamines was impaired. Additionally, the hyperlocomotor effects of NPS and methylphenidate depend on monoamines vesicular storaged, mainly dopamine, and on the activation of D2 receptors. The psychostimulant effects of NPS via activation of dopaminergic system display clinical significance on the treatment of diseases which involves dopaminergic pathways, such as Parkinson s disease and drug addiction / Neuropept?deo S (NPS) ? um pept?deo end?geno formado por 20 amino?cidos e ? o ligante de um receptor acoplado ? prote?na G chamado NPSR, o qual est? envolvido na modula??o de v?rias fun??es biol?gicas centrais como locomo??o, ansiedade, nocicep??o, ingest?o de alimento e comportamentos motivacionais. J? ? conhecido que o efeito hiperlocomotor do NPS ? mediado pelos receptores NPSR e parece depender da ativa??o do sistema adenosin?rgico, dopamin?rgico e do sistema peptid?rgico do CRF. Considerando o pouco conhecimento acerca do envolvimento do sistema dopamin?rgico na media??o do aumento da atividade locomotora induzido pelo NPS, o presente estudo objetiva investigar as a??es motoras da administra??o intracerebroventricular (icv) de NPS em camundongos pr?-tratados com α-metil-p-tirosina (AMPT, inibidor da enzima de s?ntese de dopamina), reserpina (inibidor do armazenamento da dopamina em ves?culas) ou sulpiride (inibidor de receptores D2 de dopamina), em animais submetidos ao teste de atividade locomotora no campo aberto. Camundongos Swiss machos (30-35 g) foram submetidos ? cirurgia estereot?xica para a implanta??o de uma c?nula-guia no ventr?culo lateral. No 3? dia ap?s a cirurgia, os animais foram pr?-tratados com AMPT (250 mg/kg, ip, 24 h antes do teste), reserpina (2 mg/kg, SC, 24h) ou sulpiride (25 mg/kg, ip, 45 min) e depois foram tratados com NPS (1 nmol, 2 μl; icv, 5 min) e submetidos ao teste do campo aberto. Para fins de compara??o, um grupo distinto de animais recebeu os mesmos pr?-tratamentos acima descritos (AMPT, reserpina ou sulpiride) e o efeito hiperlocomotor do metilfenidato (5 mg/kg, sc, 15 min; inibidor da recapta??o de dopamina) foi investigado no campo aberto. O teste do campo aberto avalia a locomo??o espont?nea dos animais atrav?s da dist?ncia percorrida (m) e do tempo de imobilidade (m) durante 60 min. O NPS aumentou a atividade locomotora dos animais na dose de 1 nmol. O AMPT per se n?o causou altera??o na locomo??o dos animais. Por outro lado, o AMPT reduziu parcialmente o efeito hiperlocomotor do metilfenidato, mas n?o foi capaz de afetar a a??o hiperlocomotora do NPS. Tanto o pr?-tratamento com reserpina como o com sulpiride foram capazes de inibir o efeito estimulat?rio do NPS, assim como o do metilfenidato. Estes achados mostram que o efeito hiperlocomotor do metilfenidato, mas n?o do NPS, foi afetado pela administra??o de AMPT. Al?m disso, tanto o efeito do metilfenidato quanto o do NPS foram prejudicados pelos pr?-tratamentos com reserpina e sulpiride. Em conjunto, sugere-se que o NPS pode promover est?mulo excitat?rio mesmo quando a s?ntese de catecolaminas foi prejudicada. Ainda conclui-se que o efeito hiperlocomotor do NPS e do metilfenidato depende dos estoques vesiculares de monoaminas, em particular dopamina, e da ativa??o do receptor dopamin?rgico D2. O efeito psicoestimulante do NPS por meio da ativa??o do sistema dopamin?rgico pode apresentar import?ncia cl?nica no tratamento de doen?as que envolvem a via dopamin?rgica, como o Mal de Parkinson e a depend?ncia qu?mica

京都大学 / 新制・課程博士 / 博士(薬科学) / 甲第23142号 / 薬科博第141号 / 新制||薬科||15(附属図書館) / 京都大学大学院薬学研究科医薬創成情報科学専攻 / (主査)教授 土居 雅夫, 教授 中山 和久, 教授 竹島 浩 / 学位規則第4条第1項該当 / Doctor of Pharmaceutical Sciences / Kyoto University / DFAM

An unprecedented rise in the availability of new synthetic drugs of abuse has been observed in the recent years. One of the most noted cases is that of a popularized designer drug mixture known as ‘bath salts’. Commonly obtained from various shops and on the internet, “bath salts” often contain the synthetic cathinones 3,4 methylenedioxypyrovalerone (MDPV), mephedrone, and methylone in diverse combinations. Individually, the synthetic cathinones are known to have similar pharmacology to controlled psychostimulants such as cocaine and the amphetamines, increasing the levels of dopamine (DA) in the synaptic cleft. DA is an important neurotransmitter that regulates a variety of behaviors and functions; neurons within the mesolimbic DA pathway (ventral tegmental area to nucleus accumbens) are involved in reward and motivation and are activated by these drugs of abuse. Additionally, psychostimulant-induced increases in DA in the nigrostriatal pathway (substantia nigra to corpus striatum) lead to increases in locomotor behavior. However, the majority of preclinical investigations have only assessed the effects of individual bath salt constituents and have provided little information regarding the possibility of significant drug interactions with the co-exposure of MDPV, mephedrone, and methylone. This study sought to evaluate and compare the effects of individual versus combined MDPV, mephedrone, and methylone on dopamine (DA) levels in discrete brain regions as well as motor stimulant responses in mice. Male adolescent Swiss-Webster mice received intraperitoneal injections of saline, MDPV, mephedrone, methylone (1.0 or 10.0 mg/kg), or the cathinone cocktail (MDPV + mephedrone + methylone at 1.0, 3.3, or 10 mg/kg). The effect of each treatment on DA and DA metabolite levels in mesolimbic and nigrostriatal brain tissue was quantified 15 min after a single exposure utilizing high pressure liquid chromatography with electrochemical detection (HPLC-ECD). Additionally, locomotor activity was recorded in mice after acute (day 1) and chronic intermittent (day 7) dosing. The results demonstrate that MDPV, mephedrone, and methylone produce dose-related increases in the mesolimbic and nigrostriatal DA levels that are significantly enhanced following their co-administration. Additionally, a decrease in locomotor activity on day 1 that was exacerbated by day 7 was noted in mice treated with the cathinone cocktail and was not observed with any of the single agents. The decrease in locomotor activity was accompanied by an increase in stereotypic-like behavior including excessive grooming and even self-mutilation. Our findings demonstrate a significantly enhanced effect of MDPV, mephedrone, and methylone on both DA and its metabolites resulting in significant alterations in locomotor activity. This work provides insight into the potential enhanced risk of the use of these combination synthetic cathinone products.

Repeated treatments with a dopamine (DA) D2 receptor agonist result in the induction of DA D2 receptor supersensitivity, as evidenced by enhanced behavioral responses to subsequent D2 agonist treatments - a phenomenon known as priming of receptors. Priming of D2 receptors has been well-studied in otherwise intact (non-lesioned) rats. In contrast to D2 priming, repeated treatments with a DA D1 agonist are unable to prime D1 receptors unless nigrostriatal DA fibers are largely destroyed in early postnatal ontogeny. In order to determine if D2 receptors could be primed in rats in which nigrostriatal DA fibers were largely destroyed in early postnatal ontogeny, rats were (a) lesioned at 3 days after birth with 6-hydroxydopamine (67 μg in each lateral ventricle; desipramine, 20 mg/kg IP, 1 h; 6-OHDA), (b) treated daily for the first 28 days after birth with the D2 agonist quinpirole HCl (3.0 mg/kg IP), and (c) observed in adulthood for both quinpirole-induced and SKF 38393- (D1 agonist-) induced locomotor activity and stereotyped activities. In 6-OHDA-lesioned rats in which endogenous striatal DA was reduced by 99%, quinpirole did not produce enhanced locomotor or stereotyped activities. However, SKF 38393 produced increased locomotor and stereotyped activities even after the first dose of SKF 38393. These findings demonstrate that D2 receptors are not primed by ontogenetic quinpirole treatments of neonatally 6-OHDA-lesioned rats, although D2 agonist treatments do at least partially prime D1 receptors in 6-OHDA-lesioned rats.

Designer drug mixtures popularized as “bath salts” often contain the synthetic cathinones 3,4 methylenedioxypyrovalerone (MDPV), mephedrone, and methylone in various combinations. However, most preclinical investigations have only assessed the effects of individual bath salt constituents, and little is known about whether co-exposure to MDPV, mephedrone, and methylone produces significant neuropharmacological interactions. This study evaluated and compared how MDPV, mephedrone, and methylone influence discrete brain tissue dopamine (DA) levels and motor stimulant responses in mice when administered alone and as a ternary mixture. Male adolescent Swiss-Webster mice received intraperitoneal injections of saline or 1 or 10 mg/kg doses of MDPV, mephedrone, or methylone, or a cocktail of all three cathinones at doses of 1, 3.3, or 10 mg/kg each. The effect of each treatment on DA and DA metabolite levels in mesolimbic and nigrostriatal brain tissue was quantified 15 min after a single exposure using HPLC-ECD. Additionally, locomotor activity was recorded in mice after acute (day 1) and chronic intermittent (day 7) dosing. MDPV, mephedrone, and methylone produced dose-related increases in mesolimbic and nigrostriatal DA levels that were significantly enhanced following their co-administration. In addition, mice treated with the cathinone cocktail displayed decreased locomotor activity on day 1 that was exacerbated by day 7 and not observed with any of the drugs alone. Our findings demonstrate a significant enhanced effect of MDPV, mephedrone, and methylone on both DA, and these effects on DA result in significant alterations in locomotor activity.

In many animal species, circadian rhythms of behavior are not constant throughout the lifetime of the individual but rather exhibit at least some degree of plasticity. In the present study, we have examined the potential influences of gender, age, and nutrition (presence or absence of liver) on the expression of circadian locomotor activity rhythms in the flesh fly Sarcophaga crassipalpis. We found no significant differences in endogenous circadian period under constant dark conditions with respect to gender, nutrition, or age for the duration of our experiments. On the other hand, both male and female flesh flies, as expected, were predominantly diurnal under light-dark cycles, but the pattern of entrainment differed between the sexes. Females also displayed higher activity levels than males. Also, in contrast with males, female activity levels increased with age. Moreover, females exhibited an extraordinary, but transient (one to three days), departure from diurnality which we characterize as “extended dark activity” (EDA). This phenomenon appeared as a continuous bout of locomotor activity that extended at least three hours into the early half of the dark phase at levels at least twice the median of the overall locomotor activity for the individual fly. EDA occurred as an age-dependent response to liver consumption, never appearing prior to day 4 post-eclosion but, thereafter, transpiring within one or two days after a 48-h exposure to liver. These results suggest a linkage between physiological events associated with egg provisioning and locomotor activity in the anautogenous flesh fly. Furthermore, our findings identify the existence of multiple influences on the expression of circadian clock-regulated behavior.