In order to understand how the brain functions, under normal as well as pathological conditions, it is important to study the mechanisms underlying information integration. Depending on the nature of an input arriving at a synapse, different strategies may be used by the neuron to integrate and respond to the input. Naturally, if a short train of high-frequency synaptic input arrives, it may be beneficial for the neuron to be equipped with a fast mechanism that is highly sensitive to inputs on a short time scale. If, on the contrary, inputs arriving with low frequency are to be processed, it may be necessary for the neuron to possess slow mechanisms of integration. For example, in certain working memory tasks (e. g. delay-match-to-sample), sensory inputs may arrive separated by silent intervals in the range of seconds, and the subject should respond if the current input is identical to the preceeding input. It has been suggested that single neurons, due to intrinsic mechanisms outlasting the duration of input, may be able to perform such calculations. In this work, I have studied a mechanism thought to be particularly important in supporting the integration of low-frequency synaptic inputs. It is mediated by a cascade of events that starts with activation of group I metabotropic glutamate receptors (mGlu1/5), and ends with a membrane depolarization caused by a current that is mediated by canonical transient receptor potential (TRPC) ion channels. This current, denoted ITRPC, is the focus of this thesis. A specific objective of this thesis is to study the role of ITRPC in the integration of synaptic inputs arriving at a low frequency, < 10 Hz. Our hypothesis is that, in contrast to the well-studied, rapidly decaying AMPA and NMDA currents, ITRPC is well-suited for supporting temporal summation of such synaptic input. The reason for choosing this range of frequencies is that neurons often communicate with signals (spikes) around 8 Hz, as shown by single-unit recordings in behaving animals. This is true for several regions of the brain, including the entorhinal cortex (EC) which is known to play a key role in producing working memory function and enabling long-term memory formation in the hippocampus. Although there is strong evidence suggesting that ITRPC is important for neuronal communication, I have not encountered a systematic study of how this current contributes to synaptic integration. Since it is difficult to directly measure the electrical activity in dendritic branches using experimental techniques, I use computational modeling for this purpose. I implemented the components necessary for studying ITRPC, including a detailed model of extrasynaptic glutamate concentration, mGlu1/5 dynamics and the TRPC channel itself. I tuned the model to replicate electrophysiological in vitro data from pyramidal neurons of the rodent EC, provided by our experimental collaborator. Since we were interested in the role of ITRPC in temporal summation, a specific aim was to study how its decay time constant (τdecay) is affected by synaptic stimulus parameters. The hypothesis described above is supported by our simulation results, as we show that synaptic inputs arriving at frequencies as low as 3 - 4 Hz can be effectively summed. We also show that τdecay increases with increasing stimulus duration and frequency, and that it is linearly dependent on the maximal glutamate concentration. Under some circumstances it was problematic to directly measure τdecay, and we then used a pair-pulse paradigm to get an indirect estimate of τdecay. I am not aware of any computational model work taking into account the synaptically evoked ITRPC current, prior to the current study, and believe that it is the first of its kind. We suggest that ITRPC is important for slow synaptic integration, not only in the EC, but in several cortical and subcortical regions that contain mGlu1/5 and TRPC subunits, such as the prefrontal cortex. I will argue that this is further supported by studies using pharmacological blockers as well as studies on genetically modified animals. / QC 20101005

付記する学位プログラム名: 充実した健康長寿社会を築く総合医療開発リーダー育成プログラム / 京都大学 / 0048 / 新制・課程博士 / 博士(薬科学) / 甲第21046号 / 薬科博第89号 / 新制||薬科||10(附属図書館) / 京都大学大学院薬学研究科薬科学専攻 / (主査)教授 金子 周司, 教授 竹島 浩, 教授 髙倉 喜信 / 学位規則第4条第1項該当 / Doctor of Pharmaceutical Sciences / Kyoto University / DFAM

No description available.

Das Ziel der vorliegenden Arbeit war es, die brunststimulierende Wirkung des synthetisch hergestellten Gonadorelin[5-His, 6-Asp, 7-Trp, 8-Lys] (International Nonproprietary Name: Peforelin), das in dem Präparat Maprelin® XP10 enthalten ist, bei abgesetzten pluriparen Sauen zu prüfen. Im ersten Versuchsabschnitt wurde die zweckmäßige Dosierung ermittelt. Im zweiten Versuchsabschnitt wurde die Wirkung von Maprelin® XP10 mit der einer eCG- und einer Placebobehandlung verglichen. Die Bedingungen waren für die Versuchstiere in beiden Versuchsabschnitten homogen (etwa vierter Wurf, Säugezeit vier Wochen, Brunstkontrolle zweimal täglich in Anwesenheit eines geschlechtsaktiven Ebers, zweimal täglich sonographische Ovaruntersuchung, duldungsorientierte Besamung einmal täglich). In der Dosisfindungsstudie, in die 88 Tiere einbezogen wurden, erwies sich die Dosierung von 150 µg Peforelin, 24 Stunden nach dem Absetzen appliziert, als zweckmäßig zur Brunststimulation. Andere getestete Varianten (100 µg 24 Stunden, 150 µg 0 Stunden, 150 µg 48 Stunden, 187,5 µg 24 Stunden, 255 µg 24 Stunden nach dem Absetzen) waren hierzu weniger geeignet. In der Untersuchung zur klinischen Wirksamkeit wurden die 313 einbezogenen Tiere in drei Gruppen aufgeteilt und erhielten 24 Stunden nach dem Absetzen pro Tier 150 µg Peforelin (Gruppe I), 800 IE eCG (Gruppe II) oder 2 ml physiologische NaCl-Lösung als Placebo (Gruppe III). Zur Befunderhebung an den Ovarien wurden die Tiere zweimal täglich sonographisch untersucht. Die Östrusrate nach der Peforelin-Behandlung war derjenigen nach eCG-Injektion gleichwertig (95,1 bzw. 96,3 %), beide waren der Placebobehandlung signifikant überlegen (80,6 %). In die weiteren Auswertungen wurden nur Tiere mit Brunstbeginn bis zum siebten Tag nach dem Absetzen einbezogen. Das Absetz-Östrus-Intervall betrug 100,5, 94,2 bzw. 104,1 Stunden in den Gruppen I, II bzw. III. In der Brunstdauer und dem Intervall vom Östrusbeginn bis zur Ovulation unterschieden sich die drei Gruppen nicht. Die durchschnittliche Follikelgröße war in allen drei Gruppen zum Zeitpunkt des Absetzens 4 mm und zum Zeitpunkt der ersten Duldung 6 mm. Die Ovulationen fanden sowohl nach eCG als auch nach Maprelin® XP10 zwischen dem Mittag des sechsten (13.00 h) und der Nacht des siebten Tages (1.00 h) nach dem Absetzen statt. Bei der sonographischen Trächtigkeitsuntersuchung in der vierten Woche post inseminationem waren 100,0, 99,0 bzw. 97,6 % in den Gruppen I, II bzw. III positiv. Die Trächtigkeitsrate betrug 96,9, 97,1 bzw. 91,6 % in den Gruppen I, II bzw. III. Die Abferkelrate ergab 92,2, 93,4 und 73,8 % in den Gruppen I, II und III. Das Abferkelergebnis war in allen drei Gruppen gleich (11,7, 12,0 bzw. 11,6 insgesamt geborene Ferkel in den Gruppen I, II bzw. III). Es wurde eine negative Korrelation zwischen der Dauer des Absetz-Östrus-Intervalls und der Brunstdauer bzw. dem Intervall vom Östrusbeginn bis zur Ovulation nachgewiesen. Darüber hinaus korrelierten die Brunstdauer und das Intervall Östrusbeginn bis Ovulation positiv miteinander. In allen drei Gruppen stand die Follikelgröße bei der ersten Duldung in positiver Korrelation mit der Länge des Absetz-Östrus-Intervalls. Nach längerer Säugezeit kamen die Sauen tendenziell und in der Gruppe I signifikant früher in die Brunst als nach kürzerer Laktation. Weitere überprüfte potentielle Einflussfaktoren hatten weder auf den Brunsteintritt noch auf die erzielten Wurfgrößen oder andere Parameter Auswirkungen. Mit der vorliegenden Untersuchung wurde erstmals die Wirksamkeit des synthetisch hergestellten Peforelin zur Brunststimulation bei abgesetzten pluriparen Sauen nachgewiesen. Inwieweit das Präparat für diese Indikation auch bei primiparen Sauen oder Jungsauen wirksam eingesetzt werden kann, bleibt weiteren klinischen Prüfungen vorbehalten.

Данная работа посвящена изучению и анализу уровня физической подготовленности студентов 1 курса УрФУ, занимающихся разными спортивными специализациями. В работе представлены результаты исследования уровня развития физических качеств, ориентированные на нормативы комплекса ГТО. А также рассматриваются вопросы эффективного развития физических качеств с использованием современных направлений фитнеса. / This work is devoted to the study and analysis of the level of physical fitness of first-year students of UrFU, engaged in various sports specializations. The paper presents the results of a study of the level of development of physical qualities, focused on the standards of the TRP complex. It also addresses the issues of effective development of physical qualities using modern fitness areas.

No description available.

京都大学 / 新制・課程博士 / 博士(薬学) / 甲第24564号 / 薬博第862号 / 新制||薬||243(附属図書館) / 京都大学大学院薬学研究科薬学専攻 / (主査)教授 金子 周司, 教授 竹島 浩, 教授 生田 宏一 / 学位規則第4条第1項該当 / Doctor of Pharmaceutical Sciences / Kyoto University / DFAM

Le réseau neuronal de l’hippocampe joue un rôle central dans la mémoire en modifiant de façon durable l’efficacité de ses synapses. Dans les interneurones de la couche oriens/alveus (O/A), l’induction de la potentialisation à long terme (PLT) requiert les courants postsynaptiques excitateurs évoqués par les récepteurs métabotropes du glutamate de sous-type 1a (CPSEmGluR1a) et l’entrée subséquente de Ca2+ via des canaux de la famille des transient receptor potential (TRP). Le but de ce projet était d’identifier les canaux TRP responsables des CPSEmGluR1a et d’explorer les mécanismes moléculaires régulant leur ouverture. Nous avons déterminé par des enregistrements électrophysiologiques que les CPSEmGluR1a étaient spécifiques aux interneurones O/A et qu’ils étaient indépendants de la phospholipase C. Nous avons ensuite examiné l’expression des TRPC et leur interaction avec mGluR1a par les techniques de RT-PCR, d’immunofluorescence et de co-immunoprécipitation. Nos résultats montrent que TRPC1 et mGluR1a s’associent dans l’hippocampe et que ces deux protéines sont présentes dans les dendrites des interneurones O/A. En revanche, TRPC4 ne semble s’associer à mGluR1a qu’en système recombinant et leur colocalisation paraît limitée au corps cellulaire. Finalement, nous avons procédé à des enregistrements d’interneurones dans lesquels l’expression des TRPC a été sélectivement supprimée par la transfection d’ARN interférant et avons ainsi démontré que TRPC1, mais non TRPC4, est une sous-unité obligatoire du canal responsable des CPSEmGluR1a. Ces travaux ont permis de mieux comprendre les mécanismes moléculaires à la base de la transmission synaptique des interneurones O/A et de mettre en évidence un rôle potentiel de TRPC1 dans la PLT. / The hippocampal neuronal network plays a crucial role in memory by producing long lasting changes in the efficacy of its synapses. In interneurons of stratum oriens/alveus (O/A), long term potentiation (LTP) induction requires metabotropic glutamate receptor subtype 1a (mGluR1a)-evoked excitatory postsynaptic currents (EPSCs) and subsequent Ca2+ entry through transient receptor potential (TRP) channels. The objectives of this project were to identify the TRP channels that mediate mGluR1a-evoked EPSCs and to explore molecular mechanisms that underlie their activation. Electrophysiological recordings showed that mGluR1a-evoked EPSCs were specifically observed in O/A interneurons and they were phospholipase C-independent. We then examined TRPC expression and their interaction with mGluR1a by RT-PCR, immunofluorescence and co-immunoprecipitation techniques. Our results show that TRPC1 and mGluR1a associate in hippocampus and that both proteins have overlapping distributions in dendrites of O/A interneurons. In contrast, TRPC4 seems to associate with mGluR1a only in recombinant system and their co-localization appears to be limited to the cell body. Finally, we performed recordings of interneurons in which TRPC expression was selectively suppressed by small interfering RNAs and we found that TRPC1, but not TRPC4, is an obligatory subunit of the channel that mediate mGluR1a-evoked EPSCs. This work brought new insight on molecular mechanisms underlying synaptic transmission of O/A interneurons and uncovered a potential role for TRPC1 in LTP.

Le réseau neuronal de l’hippocampe joue un rôle central dans la mémoire en modifiant de façon durable l’efficacité de ses synapses. Dans les interneurones de la couche oriens/alveus (O/A), l’induction de la potentialisation à long terme (PLT) requiert les courants postsynaptiques excitateurs évoqués par les récepteurs métabotropes du glutamate de sous-type 1a (CPSEmGluR1a) et l’entrée subséquente de Ca2+ via des canaux de la famille des transient receptor potential (TRP). Le but de ce projet était d’identifier les canaux TRP responsables des CPSEmGluR1a et d’explorer les mécanismes moléculaires régulant leur ouverture. Nous avons déterminé par des enregistrements électrophysiologiques que les CPSEmGluR1a étaient spécifiques aux interneurones O/A et qu’ils étaient indépendants de la phospholipase C. Nous avons ensuite examiné l’expression des TRPC et leur interaction avec mGluR1a par les techniques de RT-PCR, d’immunofluorescence et de co-immunoprécipitation. Nos résultats montrent que TRPC1 et mGluR1a s’associent dans l’hippocampe et que ces deux protéines sont présentes dans les dendrites des interneurones O/A. En revanche, TRPC4 ne semble s’associer à mGluR1a qu’en système recombinant et leur colocalisation paraît limitée au corps cellulaire. Finalement, nous avons procédé à des enregistrements d’interneurones dans lesquels l’expression des TRPC a été sélectivement supprimée par la transfection d’ARN interférant et avons ainsi démontré que TRPC1, mais non TRPC4, est une sous-unité obligatoire du canal responsable des CPSEmGluR1a. Ces travaux ont permis de mieux comprendre les mécanismes moléculaires à la base de la transmission synaptique des interneurones O/A et de mettre en évidence un rôle potentiel de TRPC1 dans la PLT. / The hippocampal neuronal network plays a crucial role in memory by producing long lasting changes in the efficacy of its synapses. In interneurons of stratum oriens/alveus (O/A), long term potentiation (LTP) induction requires metabotropic glutamate receptor subtype 1a (mGluR1a)-evoked excitatory postsynaptic currents (EPSCs) and subsequent Ca2+ entry through transient receptor potential (TRP) channels. The objectives of this project were to identify the TRP channels that mediate mGluR1a-evoked EPSCs and to explore molecular mechanisms that underlie their activation. Electrophysiological recordings showed that mGluR1a-evoked EPSCs were specifically observed in O/A interneurons and they were phospholipase C-independent. We then examined TRPC expression and their interaction with mGluR1a by RT-PCR, immunofluorescence and co-immunoprecipitation techniques. Our results show that TRPC1 and mGluR1a associate in hippocampus and that both proteins have overlapping distributions in dendrites of O/A interneurons. In contrast, TRPC4 seems to associate with mGluR1a only in recombinant system and their co-localization appears to be limited to the cell body. Finally, we performed recordings of interneurons in which TRPC expression was selectively suppressed by small interfering RNAs and we found that TRPC1, but not TRPC4, is an obligatory subunit of the channel that mediate mGluR1a-evoked EPSCs. This work brought new insight on molecular mechanisms underlying synaptic transmission of O/A interneurons and uncovered a potential role for TRPC1 in LTP.

Made available in DSpace on 2015-05-14T13:00:13Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2275378 bytes, checksum: fcb4196c739d83997818dc074daf738f (MD5) Previous issue date: 2010-03-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The pharmacological effects of carvacrol, a monoterpenoid phenol, on the cardiovascular system were studied in normotensive rats, using in vivo and in vitro techniques. In superior mesenteric artery rings isolated from rats with functional endothelium carvacrol (10-8 - 3 ₓ 10-4 M) concentration-dependently relaxed phenylephrine-induced contractions (pD2 = 4.59  0.02, MR = 103.03  1.5%, N=8) and this effect was not altered after removal of the endothelium (pD2 = 4.36  0.02, MR = 111.03  4.8%, N=8), suggesting that the vasorelaxant response induced by carvacrol appears to be independent of vascular endothelium. Furthermore, carvacrol antagonized the vasoconstriction induced by high K+ solution (Tyrode with 80 mM of KCl) (pD2 = 4.12  0.01, MR = 94.38  3.97%, N=6), inhibited contraction elicited by CaCl2 in depolarizing (KCL 60 mM) nominally without Ca2+ medium out carvacrol also antagonized the contractions induced by the L-type Ca2+ channel agonist, S(-)-Bay K 8644 (pD2 = 4.537  0.023, MR = 9.8  3.58%, N=6), indicating that the vasodilatation involve probably the inhibition of Ca2+ influx through L-type voltage-dependent calcium channels (Cav type-L). Additionally, carvacrol antagonized the contractions induced by CaCl2 in nominally without Ca2+ medium in the presence of PHE and nifedipine, suggesting a possible inhibition of calcium influx by store operated channels (SOC), receptor operated channels (ROC) and/or TRP channels. Interestingly, in a depolarizing (KCL 60 mM) nominally without Ca2+ medium and in the presence of nifedipine, carvacrol also inhibited the contraction induced by CaCl2, suggesting a probable inhibition of SOC and/or TRP channels. To evaluate the involvement of TRP channels in the vasorelaxant effect induced by carvacrol, non-selective inhibitors were used. No change in the relaxation response was observed in the presence of ruthenium red (pD2= 4.31  0.029, N=6), however, the effect induced by carvacrol was potentiated by La3+ (pD2 = 5.231  0,04, N=6), Gd3+ (pD2 = 4.97  0.02, N=6) or Ni2+ (pD2 = 5.079  0.02, N=6), furthermore, Mg2+ (pD2 = 4.168  0.021; MR = 81.12  4.03%, N=6) attenuated the relaxation elicited by carvacrol, suggesting that monoterpenoid may to action on TRPC1, TRPC3, TRPC6 and TRPM7 channels. Carvacrol also induced hypotension and bradycardia in non-anesthetized normotensive rats. In conclusion, these results suggest that carvacrol induced vasorelaxant effect in superior mesenteric artery rats isolated probably inhibiting Ca2+ influx by Cav, SOC (TRPC1), ROC (TRPC1 or TRPC6) and TRPM7 channels. Moreover, the effects induced by carvacrol in normotensive non-anesthetized rats showed a hypotensive and bradycardic activity. / Os efeitos farmacológicos de carvacrol, um fenol monoterpenóide, sobre o sistema cardiovascular, foi estudado em ratos normotensos, usando técnicas in vivo e in vitro. Carvacrol (10-8 - 3 ₓ 10-4 M) induziu vasorelaxamento dos anéis de artéria mesentérica superior isolada de rato pré-contraídos com 10 μM FEN (pD2 = 4,59  0,02, Emáx = 103,03  1,5%) na presença do endotélio funcional e esse efeito não foi alterado após a remoção do endotélio (pD2 = 4,36  0,02, Emáx = 111,03  4,8%), sugerindo, portanto, que a resposta vasorelaxante induzida por carvacrol parece ser independente do endotélio vascular. Interessantemente em anéis pré-contraídos com KCl 80 mM (pD2 = 4,12  0,01, Emáx = 94,38  3,97%), observou-se uma diminuição na sua potência e na sua eficácia farmacológica, sugerindo um passo comum na via que seria um aumento citosólico dos níveis de cálcio. Adicionalmente, carvacrol antagonizou, de maneira dependente de concentração, as contrações induzidas por CaCl2 em meio despolarizante nominalmente sem Ca2+ e induziu relaxamento das contrações induzidas pelo S(-)-Bay K 8644 (pD2 = 4,537  0,023, Emáx = 91,8  3,58%) com uma diminuição na sua eficácia farmacológica, sugerindo uma inibição do influxo de cálcio por canais de Ca2+ tipo-L. Além disso, antagonizou as contrações induzidas por CaCl2 em meio nominalmente sem cálcio, na presença de FEN e nifedipina, sugerindo uma provável inibição do influxo de cálcio por SOC, ROC e/ou canais TRP. Como também, em um meio despolarizante e nominalmente sem cálcio na presença de nifedipina esse mesmo antagonismo foi observado, ressaltando a provável inibição dos SOC e/ou canais TRP. Para avaliar a participação dos canais TRP, as preparações foram incubadas com La3+ (pD2 = 5,231  0,04) , Gd3+ (pD2 = 4,97  0,02) e Ni2+ (pD2 = 5,079  0,02) onde seu efeito foi potencializado sugerindo sua ação sobre os canais TRPC e ao utilizar magnésio (pD2 = 4,168  0,021 e Emáx = 81,12  4,03%) tanto sua potência quanto sua eficácia farmacológica foi atenuada, sugerindo inibição do canal TRPM7. Nos estudos in vivo, em ratos normotensos não anestesiados, carvacrol produziu hipotensão e bradicardia. Em conclusão, esses resultados sugerem que carvacrol induz efeito vasorelaxante em anéis de artéria mesentérica superior isolada de rato por inibir provavelmente TRPM7, como também inibir o influxo de cálcio por Cav, SOC, ROC e ou TRPC1 e 6. Além disso, os efeitos induzidos por carvacrol em ratos normotensos não anestesiados mostrou uma atividade hipotensora e bradicárdica.