京都大学 / 新制・論文博士 / 博士(医学) / 乙第13441号 / 論医博第2240号 / 新制||医||1054(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 生田 宏一, 教授 滝田 順子, 教授 江藤 浩之 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

This work examines somatosensory processing in "giant" neurones of the medial pontomedullary reticular formation (PMRF) in the rat, with particular emphasis on the response to cutaneous thermal stimuli. Thermal test stimuli were employed as these were deemed to be more precisely quantifiable than other forms of cutaneous stimulation. Activity was recorded from 235 PMRF neurones in 94 female Long Evans rats (270 to 320 g) anaesthetised with urethane (1,25g/kg, i.p.). Rectal temperature was closely controlled at 38 ± 0,5°C. Standard stereotactic and extracellular recording techniques were employed. PMRF giant neurones were identified by their stereotactic location, large, stable spike amplitudes of long duration, responses to cutaneous mechanical stimuli and receptive field properties, and spontaneous discharge characteristics. Ramp, step and sine wave cutaneous thermal stimuli (35-48 °C) were applied to the glabrous skin on the hindpaw by means of a computer-controlled Peltier device. The location of the units was confirmed by subsequent histology. One hundred and eleven neurones were located in nucleus reticularis pontis caudalis (NPC), and 124 in nucleus reticularis gigantocellularis (NGC). Mechanical stimulation excited 188 of 235 (80%) PMRF neurones (ON-m cells), and inhibited 40 (17%, OFF-m cells). Seven cells (3%) had mosaic receptive fields of excitation and inhibition (complex responses, CX-m). Twenty-eight percent of neurones were responsive to both weak and intense stimuli (mixed neurones). The remainder (72%) responded only to intense mechanical stimulation of the skin (high threshold neurones). The (excitatory or inhibitory) response of the mixed neurones to intense stimuli was generally greater than to mild stimuli, Receptive fields ranged in size from restricted (hindlimbs only) to very extensive (covering the entire body surface). Neurones with small receptive fields were almost exclusively of the high threshold type, and tended to be located in NGC, while mixed neurones tended to have larger receptive fields, and were located predominantly in NPC. Some portion of the hind limbs were represented in the receptive fields of all but one of the neurones studied, while the tail and/ or trunk were represented in 77%, and the forelimbs and face in 28% of receptive fields. Most of the cells responding to cutaneous mechanical stimulation had bilateral (usually symmetric) receptive fields. Spontaneous (background) activity occurred in the absence of any deliberate sensory stimulation in 72% of PMRF neurones. The frequency of spontaneous discharge rates ranged from O to 47 spikes/ s. The coefficient of variation of the spontaneous discharge rate of a given neurone was generally less than 20% (range O to 85%). Of the 235 identified mechanosensitive PMRF neurones, 203 (86%) also responded to cutaneous thermal stimulation (43-48 °C) of the ipsilateral hind paw. Eighty percent of these responded with increased discharge rates (ON-t cells), and 20% were inhibited (OFF-t cells). The polarities of response of individual PMRF neurones to mechanical and thermal stimuli, and to repeated ipsilateral and contralateral thermal stimuli, did not differ significantly. Following transient thermal stimulation, spontaneous discharge rates largely returned to pre-stimulus levels. The thresholds of response to slow ramp (0,15°C/s) and stepped (2°C/s) thermal stimuli occurred both in the innocuous and noxious temperature ranges (below and above 42°C, respectively). The threshold temperatures showed large variability to repeated identical thermal stimuli. Despite the poor reproducibility of the threshold responses, the distribution of thresholds to thermal ramp stimuli was consistently bimodal, with peaks occurring at 39 and 43°C. The bimodality persisted even when the ipsilateral and contralateral data were pooled. The modes of these threshold distributions conform to the maximum discharge ranges for warm and noxious cutaneous receptors. Thus, it is likely that thermal input to individual PMRF neurones is derived from both types of receptors. The responses of PMRF neurones to repeated thermal stimuli were stable and reproducible with respect to magnitude and time course. The average (static) and maximum (dynamic) responses to thermal stimuli were generally small: for example, the mean of the average responses to ramp stimuli was 5,9 spikes/s ± 11,0 SD, (range -28 to 40 spikes/s), and the mean of the maximum responses was 9,3 spikes/s ± 16,1 SD, (range -46 to 65 spikes/s). The absolute change in firing rate of individual PMRF neurones, and of the population, increased monotonically as a function of the intensity of stepped cutaneous thermal stimuli in the range 40 to 48 °C. However, their resolution, based on their average and maximum responses, was poor. Incorporating the post-stimulus responses into the comparisons between different stimulus intensities marginally increased the resolution of these neurones. Thus, while the majority of PMRF neurones are able to distinguish innocuous from noxious stimuli, few are capable of encoding stimulus intensity within the noxious range (above 43 °C). The majority (70%) of PMRF neurones responded to sustained thermal stimuli with a slow increase or decrease to a new static discharge rate which was maintained with little or no adaptation. Latency to onset of response to stepped thermal stimuli varied from 1 to 50 seconds, and the time to maximal response between 5-60 seconds. Many PMRF neurones also showed marked after-discharge for periods of up to 5 minutes after removal of the stimulus. The thermal receptive fields of over 90% of PMRF neurones were large, incorporating at least both hindlimbs. The extensive receptive field sizes of individual PMRF neurones provides evidence against them having a role in stimulus location. The large number of PMRF neurones showing multimodal convergence, their small magnitude responses, their slow response times, and their large receptive fields strongly suggest that these neurones are not participating in classical sensory discrimination. Rather, they may function as stimulus detectors or alternatively play a role in associative processes.

Abstract: Metal-Organic Frameworks (MOFs) are a class of highly porous, hybrid, functional and crystalline extended coordination compounds. Their exceptional properties renders them ideal for a wide range of applications including gas storage and catalysis. Especially for catalysis, MOFs are receiving attention as well-defined supports for organometallic heterogeneous catalysis with noticeably the post-synthetic grafting of transition metal complexes on secondary building units (SBU) containing hydroxides moieties. The objective of this dissertation is to explore the synthesis, reactivity and functionalization of MOFs with SBU containing hydroxides units by transition metal catalyst using the Surface Organometallic Chemistry (SOMC) approach. Chapter 1, gives an introduction to the field of MOF and their applications to catalysis through the functionalization of hydroxide containing SBUs. This chapter introduces also the SOMC strategy with an overview of its catalytic application for olefin metathesis and CO2 conversion. Chapter 2 and 3 give a detailed application of SOMC to MOFs with the selective grafting of the W(≡CtBu)(CH2tBu)3 complex on the highly crystalline and mesoporous Zr-NU-1000 MOF. The obtained single site material, Zr-Nu-1000-W, is fully characterized using state of the art experimental methods and all the steps leading to the final grafted moieties were identified by DFT. Zr-NU-1000-W is active for olefin metathesis and is further fine-tuned by activation with EtAlCl2 giving a more selective and stable catalyst. Moreover, the nature of the grafted species could be modulated by pre-activation of the initial W(≡CtBu)(CH2tBu)3 complex with dmpe giving W(≡CtBu)(=CHtBu)(CH2tBu)(dmpe) also grafted on Zr-NU-1000. Chapter 4 and 5, describe the deliberate design and bulk synthesis of a new zirconium MOF, Zr-she-MOF-2, and highlight the discovery of a new highly connected MOF, RE-urx-MOF-1, based on a careful combination of rare earth (RE) metals with heterobifunctional triangular tetrazolate-based ligand. Additionally, the replacement of the tetrazolate functionality by carboxylate, leads to the formation of a different MOF structure RE-gea-MOF-4 having the gea topology with the presence of 18-connected nonanuclear RE cluster. Both Zr-she-MOF-2 and RE-gea-MOF-4 are active for the coupling of epoxides with CO2 to form cyclic carbonate in the presence of Bu4NBr. Finally, Chapter 6 will discuss the conclusions and perspectives of this dissertation.

Metastin, the gene product of metastasis suppressor gene KiSS-1, is the endogenous ligand for the G-protein-coupled receptor GPR54 (or AXOR12, or OT7T175). The expression of KiSS-1 gene and peptide and the distribution of metastin were studied in the rat central nervous system by reverse transcriptase-polymerase chain reaction, Western blotting, and immunohistochemical methods. KiSS-1 gene and peptide expression was higher in the hypothalamus than in the brainstem and spinal cord. In the brain, metastin-like immunoreactivity (irMT) was found mainly in three groups of cells: dorsomedial hypothalamic nucleus, nucleus of the solitary tract, and caudal ventrolateral medulla. Immunoreactive fibers of varying density were noted in bed nucleus of stria terminalis, septal nuclei, nucleus accumbens, caudate putamen, diagonal band, amygdala, hypothalamus, zona incerta, thalamus, periaqueductal gray, raphe nuclei, lateral parabrachial nucleus, locus coeruleus, spinal trigeminal tract, rostral ventrolateral medulla, and medullary reticular nucleus. Preabsorption of the antiserum with metastin peptide fragment (45-54)-NH2 (1 μg/ml) resulted in no staining in any of the sections. The biological activity of metastin was assessed by monitoring intracellular calcium [Ca2+]i in cultured hippocampal neurons, which are known to express GPR54. Metastin increased [Ca 2+]i in a population of cultured hippocampal neurons. The results show that metastin is biologically active in rat central neurons, and its anatomical distribution suggests a possible role in nociception and autonomic and neuroendocrine functions.

Final publication is available at http://www.sciencedirect.com/science/article/pii/S0002939414000488 / 京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19586号 / 医博第4093号 / 新制||医||1014(附属図書館) / 32622 / 京都大学大学院医学研究科医学専攻 / (主査)教授 大森 孝一, 教授 宮本 享, 教授 横出 正之 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Distribution of prolactin-releasing peptide-like immunoreactivity (PrRP- LI) was investigated in the rat medulla with the use of a rabbit polyclonal antiserum against the human PrRP-31 peptide. PrRP-positive neurons were noted mainly in two areas of the caudal medulla: ventrolateral reticular formation and commissural nucleus of the nucleus of the solitary tract (NTS), corresponding to the A1 and A2 areas. PrRP-LI neurons were absent in the medulla rostral to the area postrema. Double-labeling the sections with PrRP antisera and tyrosine hydroxylase (TH) monoclonal antibodies revealed extensive colocalization of PrRP- and TH-like immunoreactivity (TH-LI) in neurons of the A1 and A2 areas. Our results show that PrRP-LI is expressed in a population of A1 and A2 noradrenergic neurons of the rat caudal medulla.

Projections from medial agranular cortex to brain stem in rat were determined by use of the anterograde tracers Phaseolus vulgaris leucoagglutinin, or wheat germ agglutinin conjugated horseradish peroxidase. Axonal trajectories were also followed by means of the Wiitanen modification of the Fink-Heimer degeneration technique. AGm was identified on the basis of its cytoarchitectonics. AGm projected to the anterior pretectal nucleus, the rostral interstitial nucleus of the medial longitudinal fasciculus, the medial accessory oculomotor nucleus of Bechterew, the interstitial nucleus of Cajal, the nucleus of Darkschewitsch, the nucleus cuneiformis and subcuneiformis, intermediate and deep superior collicular layers, the paramedian pontine reticular formation (reticularis pontis oralis and caudalis, and reticularis gigantocellularis), and raphe centralis superior. Differences in connections between rostral and caudal injections were observed: pontine and medullary projections were lighter from the rostral portion of AGm than from the more caudal portions of AGm. The heaviest projections to the anterior pretectal nucleus were from the caudal portion of AGm. The subcortical projections were very similar to those described for the frontal eye field in monkeys, and the majority of them targeted areas thought to be involved in coordination of gaze with head and neck movements. Thus AGm in rats may contain the homologue of the primate frontal eye fields.

The rational design and construction of Metal-Organic Frameworks (MOFs) with intricate structural complexity are of prime importance in reticular chemistry. However, the design of intricate structures that can practically be synthesized is very difficult, and the suitable targeted intricate nets are still unexplored. Evidently, it is of great value to build the fundamental theory for the design of intricate structures. This dissertation is focused on the exploration of cutting-edge design methodologies in reticular chemistry. This research shows the design and synthesis of several MOF platforms (hex, fcu, gea and the) based on rare earth polynuclear clusters. Furthermore, this research unveils the latest addition, named merged nets approach, to the design toolbox in reticular chemistry for the rational design and construction of intricate mixed-linker MOFs. In essence, a valuable net for design enclosing two edges is rationally generated by merging two edge-transitive nets, spn and hxg. The resultant merged net, named sph net, offers potential for the deliberate design and construction of highly symmetric isoreticular intricate mixed-linker MOFs, sph-MOF-1 to 4, which represent the first examples of MOFs where the underlying net is merged from two 3-periodic edge-transitive nets. Furthermore, the underlying principle of the merged net approach, the fundamental merged net equation, and two key parameters are disclosed. Also, we discovered three analysis methods to check and validate corresponding signature nets in an edge-transitive net. Based on these analysis methods, a signature map of all edge-transitive nets was established. This map showing the systematic relationship among edge-transitive nets will help the material chemist to comprehend more about the underlying nets in reticular chemistry. Based on the revealed map, we systematically described the nine types of merging combination and 140 merged nets based on two edge-transitive nets. Among these enumerated nets, only 18 of them was shown on the RCSR database before. These enumerated merged nets significantly increased the designable targets in reticular chemistry. Using an example of enumerated sub net, we show how this approach can be utilized to design and synthesis mixed-linker porous materials based on the intricate sub-MOF platform, which presents one of the most intricate MOF structures synthesized by design.

No description available.

La restructuration productive de l’industrie pauliste survenue à la fin des années 1980 a réservé une place importante à la main d’œuvre migrante dans le secteur de la confection. Le circuit de sous-traitance transnational a favorisé l’arrivée des travailleurs mobiles venus des pays voisins. Suite à une intensification de ce flux migratoire au tournant des années 2000, les Boliviens se sont vite imposés comme la plus importante population migrante latino-américaine de la ville de São Paulo. Ils travaillent dans des ateliers de confection, de petites structures de production ayant l’avantage de faciliter la mobilité sociale et spatiale grâce à des carrières d’entrepreneurs migrants. Les conditions de production de l’espace géographique et social de cette migration sont mises en perspective par une identification des réseaux sociaux dont dépendent les trajectoires migratoires. Afin de saisir l’articulation entre réseaux et mobilités, nous partons du constat de l’existence de dispositifs réticulaires aux agencements variables qui opèrent comme des matrices des migrations individuelles et participent à la production d’un espace social transnational. Ces dispositifs réticulaires sont basés soit sur des liens jetables, soit sur des liens familiaux, soit sur des liens de communauté rurale. Ils donnent à voir des compétences circulatoires et relationnelles engageant une capacité d’action chez les individus concernés qui contraste avec la représentation du « travailleur esclave » qui structure une partie du rapport entre les pouvoirs publics et les migrants boliviens. / The productive restructuring of the paulist industry in the late 1980s has reserved an important place to the migrant labour in the garment sector. The transnational subcontracting circuit has favoured the arrival of mobile workers from neighbouring countries. In the wake of an intensification of this migratory flow at the turn of the 2000s, Bolivians has quickly established themselves as the largest Latin American migrant population in the city of São Paulo. They work in sweatshops, small production structures having the advantage of facilitating social and spatial mobility through careers as migrant entrepreneurs. The conditions of production of the geographical and social space of this migration are put into perspective by identifying the social networks on which the migratory trajectories depend. In order to understand the articulation between networks and mobilities, we start from the observation of the existence of reticular apparatus with variable arrangements that operate as individual migrations matrices and participate in the production of a transnational social space. These reticular apparatus are based either on disposable ties or on family ties or on rural community ties. They reveal circulatory and relational skills involving a capacity for action among the individuals concerned, which contrasts with the representation of "slave labourer" that structure part of the relationship between government and Bolivian migrants.