Rats lesioned shortly after birth with 6-hydroxydopamine (6-OHDA; 134 μg icv) represent a near-ideal model of severe Parkinson's disease because of the near-total destruction of nigrostriatal dopaminergic fibers. The element manganese, an essential cofactor for many enzymatic reactions, itself in toxic amount, replicates some clinical features similar to those of Parkinson's disease. The aim of this study was to examine the effect of neonatal manganese exposure on 6-OHDA modeling of Parkinson's disease in rats. Manganese (MnCl 2·4H 2O) 10,000 ppm was included in the drinking water of pregnant Wistar rats from the time of conception until the 21st day after delivery, the age when neonatal rats were weaned. Control rats consumed tap water. Other groups of neonatal rat pups, on the 3rd day after birth, were pretreated with desipramine (20 mg/kg ip 1 h) prior to bilateral icv administration of 6-OHDA (30, 60, or 137 μg) or its vehicle saline-ascorbic (0.1%) (control). At 2 months after birth, in rats lesioned with 30, 60, or 134 μg 6-OHDA, endogenous striatal dopamine (DA) content was reduced, respectively, by 66, 92, and 98% (HPLC/ED), while co-exposure of these groups to perinatal manganese did not magnify the DA depletion. However, there was prominent enhancement of DA D 1 agonist (i.e., SKF 38393)-induced oral activity in the group of rats exposed perinatally to manganese and also treated neonatally with the 30 mg/kg dose of 6-OHDA. The 30 mg/kg 6-OHDA group, demonstrating cataleptogenic responses to SCH 23390 (0.5 mg/kg) and haloperidol (0.5 mg/kg ip), developed resistance if co-exposed to perinatal manganese. In the group exposed to manganese and lesioned with the 60 mg/kg dose of 6-OHDA, there was a reduction in D2 agonist (i.e., quinpirole, 0.1 mg/kg)-induced yawning. The series of findings demonstrate that ontogenetic exposure to manganese results in an enhancement of behavioral toxicity to a moderate dose of 6-OHDA, despite the fact that there is no enhanced depletion of striatal DA depletion by the manganese treatment.

To assess the possible modulatory effects of noradrenergic and serotoninergic neurons on dopaminergic neuronal activity, the noradrenergic and serotoninergic neurotoxins DSP-4 N-(2-chlorethyl)-N-ethyl-2-bromobenzylamine (50.0 mg/kg, sc) and 5,7- dihydroxytryptamine (5,7-DHT) (37.5 μg icv, half in each lateral ventricle), respectively, were administered to Wistar rats on the first and third days of postnatal ontogeny, and dopamine (DA) agonist-induced behaviors were assessed in adulthood. At eight weeks, using an HPLC/ED technique, DSP-4 treatment was associated with a reduction in NE content of the corpus striatum (> 60%), hippocampus (95%), and frontal cortex (> 85%), while 5,7-DHT was associated with an 80-90% serotonin reduction in the same brain regions. DA content was unaltered in the striatum and the cortex. In the group lesioned with both DSP-4 and 5,7-DHT, quinpirole-induced (DA D2-agonist-agonist) yawning, 7-hydroxy-DPAT-induced (DA D3 agonist) yawning, and apomorphine-induced (non-selective DA agonist) stereotypies were enhanced. However, SKF 38393-induced (DA D1 agonist) oral activity was reduced in the DSP-4 + 5,7-DHT group. These findings demonstrate that DA D2- and D3-agonist-induced behaviors are enhanced while DA D1-agonist-induced behaviors are suppressed in adult rats in which brain noradrenergic and serotoninergic innervation of the brain has largely been destroyed. This study indicates that noradrenergic and serotoninergic neurons have a great impact on the development of DA receptor reactivity (sensitivity).

In the neonatally 6-hydroxydopamine (6-OHDA)-lesioned rat hyperlocomotor activity, first described in the 1970s, was subsequently found to be increased by an additional lesion with 5,7-dihydroxytryptamine (5,7-DHT) (i.c.v.) in adulthood. The latter animal model (i.e., 134 μg 6-OHDA at 3 d postbirth plus 75 μg 5,7-DHT at 10 weeks; desipramine pretreatments) was used in this study, in an attempt to attribute hyperlocomotor attenuation by D,L-amphet-amine sulfate (AMPH) and m-chlorophenylpi-perazine di HCl (mCPP), to specific changes in extraneuronal (i.e., in vivo microdialysate) levels of dopamine (DA) and/or serotonin (5-HT). Despite the 98-99% reduction in striatal tissue content of DA, the baseline striatal microdialysate level of DA was reduced by 50% or less at 14 weeks, versus the intact control group. When challenged with AMPH (0.5 mg/kg), the microdialysate level of DA went either unchanged or was slightly reduced over the next 180 min (i.e., 20 min sampling), while in the vehicle group and 5,7-DHT (alone) lesioned group, the microdialysate level was maximally elevated by ∼225% and ∼450%, respectively - and over a span of nearly 2 h. Acute challenge with mCPP (1 mg/kg salt form) had little effect on microdialysate levels of DA, DOPAC and 5-HT. Moreover, there was no consistent change in the microdialysate levels of DA, DOPAC, and 5-HT between intact, 5-HT-lesioned rats, and DA-lesioned rats which might reasonably account for an attenuation of hyperlocomotor activity. These findings indicate that there are other important neurochemical changes produced by AMPH-and mCPP-attenuated hyperlocomotor activity, or perhaps a different brain region or multiple brain regional effects are involved in AMPH and mCPP behavioral actions.

京都大学 / 新制・課程博士 / 博士(農学) / 甲第23522号 / 農博第2469号 / 新制||農||1087(附属図書館) / 学位論文||R3||N5353(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 小杉 緑子, 教授 北山 兼弘, 教授 柴田 昌三 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

As a part of the overall program in the Grandy laboratory at Oregon Health & Science University (OHSU), studying the underlying chemical biology of methamphetamine (Meth) addiction, this dissertation reports on the development of six new thyronamine analogs which were synthesized and assayed against trace amine associated receptor 1 (TAAR1), giving preliminary results consistent with the analogs being inverse agonists. Due to highly variable TAAR1 expression levels in the assays, based on inter-assay response to control Meth stimulation as well as other possible factors, kinetic models were developed to qualitatively explain the assay results. The models set approximate limits on the analogs' binding and disassociation rates relative to those of Meth. Analysis of the assays also provides more evidence of TAAR1's basal activity. Based on the models, the conversion rate of ligand-free inactive TAAR1 to ligand-free active TAAR1 is less than 6% of the binding rate of Meth to TAAR1. The models also suggest that the inverse agonists bind to the inactive ligand-free form of TAAR1 between 10 and 100 times faster than Meth binds to the inactive ligand-free form of TAAR1. Three of the new analogs, G5-110s8, G5-112s5, and G5-114s5, bind to the ligand-free active form of TAAR1 faster than they bind to the inactive ligand-free form of TAAR1. The models do not suggest an upper limit on the binding rate of those 3 analogs to the ligand-free active form of TAAR1. A control assay lacking TAAR1 revealed an electrophysiological off-target effect caused by G5-109s8. Also, a novel synthetic route was developed for ET-92, the lead compound for this project, which reduced the number of synthetic steps from 14 to 5 and improved the overall yield from 15.3% to 18.3% (77.4 mg) with the hope that further improvements in yield are possible.

京都大学 / 0048 / 新制・課程博士 / 博士(地球環境学) / 甲第20038号 / 地環博第154号 / 新制||地環||31(附属図書館) / 33134 / 京都大学大学院地球環境学舎環境マネジメント専攻 / (主査)教授 梶井 克純, 准教授 真常 仁志, 准教授 田中 周平 / 学位規則第4条第1項該当 / Doctor of Global Environmental Studies / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第20925号 / 理博第4377号 / 新制||理||1628(附属図書館) / 京都大学大学院理学研究科地球惑星科学専攻 / (主査)教授 田上 高広, 教授 下林 典正, 准教授 河上 哲生 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM

During the latter half of the 20th century until present day there has been an unprecedented rise in global annual mean temperatures accompanied by rising sea levels and a decrease in Northern Hemisphere snow cover, which if it continues will lead to widespread disruption of climate patterns, ecosystems, and present-day landscapes. It is therefore of critical importance to establish an expanded network of paleoclimate records across the globe in order to better assesses how the global climate system has changed in the past, that we may create a metric by which to address modern change. Herein is presented a7,700 years record of Holocene climatic and environmental variability in Sermilik Valley, located on Ammassalik Island, SE Greenland. This objective of this study is to determine the timing of major Holocene climate transitions as expressed in the physical, elemental, and geochemical parameters preserved in the 484 cm sediment record of Lower Sermilik Lake. Major transitions observed in this study include the deglaciation of Sermilik Valley, the onset and termination of the Holocene Climatic Optimum, the transition into neoglacial conditions, and the Little Ice Age.

The biogenic amines serotonin (5HT), dopamine (DA), and octopamine (OA) have been indicated in the regulation of behaviors, including aggression. The flesh fly, Sarcophaga crassipalpis, was used to investigate ontogenetic and circadian changes in amines and aggression. Heads of male flies were analyzed for amine content using high performance liquid chromatography with electrochemical detection (HPLC-ECD) at 3 time points on each of 4 consecutive days in 2 light cycles, 12:12 LD and 15:9 LD. Both DA and OA levels decreased with age. Light-cycle dependent differences were observed for all amines in overall levels and patterns of change throughout the day. A behavioral assay quantified interactive and aggressive behaviors at three time points in the light period for 2 age groups. The daily changes in behavioral profiles differed dependent on age. Correlations from these data can be made between changes in amine levels and time of day, photoperiod, age, and aggressive state.

Widely regarded as a keystone species and ecosystem engineer, the eastern oyster Crassostrea virginica plays a vital role in estuarine environments. Complex, three-dimensional oyster reefs act as havens for biodiversity and contribute to ecological processes. Recently, concern for this resource has arisen in Mosquito Lagoon, Florida, the southernmost limit along the Atlantic coast for undisturbed, intertidal reefs of C. virginica. Since the 1990s, intense recreational boating activity has caused atypical dead margins (mounds of disarticulated shells) to emerge on the seaward edges of oyster reefs located along major navigational channels. Once dead margins are formed, little is known about their influence on biotic composition and interactions on oyster reefs. This study focused on the affect of dead margins on: (1) mobile species biodiversity and distribution, (2) reef architecture, and (3) the affect of structural variables on predation of juvenile oysters. To determine if dead margins influenced the biodiversity of mobile species on oyster reefs, lift nets (1 m2) were deployed within Mosquito Lagoon for one year (June 2004 - June 2005). These nets (5/site) were deployed on the back-reef areas of six reefs (3 reference reefs and 3 reefs containing dead margins). To simulate reef habitat, one and a half liters of live oysters were placed within each net. Lift nets were checked monthly and surveyed for all mobile species. The resulting data were assimilated into a species inventory containing 65 species of fishes, mollusks, crustaceans, worms, and echinoderms. The two most abundant species present on reefs in Mosquito Lagoon were the big-claw snapping shrimp Alpheus heterochaelis, a filter-feeder, and the flat-back mud crab Eurypanopeus herbstii, a predator of oyster spat. Contrary to expections, analyses of community metrics showed that dead margins did not significantly affect the biodiversity of back-reef areas on oyster reefs. Modified lift nets (0.25 m2) were placed on six different oyster reefs (3 reference reefs and 3 containing dead margins) to test if dead margins affected the distribution of mobile species inhabiting oyster reefs. Nine nets were arranged to cover three separate areas of each reef: the fore-reef (3 nets), mid-reef (3 nets), and back-reef (3 nets). Half a liter of oyster shells were placed inside each net. These nets were checked weekly, for five weeks and species richness, density, and biomass were recorded. Analyses revealed that all community metrics were significantly higher on reference reefs than reefs affected with dead margins. Further, a significant drop in all three metrics was seen on the mid-reef area of affected reefs. The absence of species on this area is hypothesized to be due to a lack of water, shade, and habitat complexity. To document architectural differences, two types of transects were run along five reference reefs and five reefs with dead margins. First, quadrat transects determined the percent of live oysters, the percent of shell clusters, topographic complexity (using chain links), and the angle of shells on each reef type. Transect lines were stretched parallel to the water line and covered all three reef areas (fore-reef, mid-reef, and back-reef). The results showed reference reefs to have approximately four-fold more live oysters, approximately twice as many shell clusters, and significantly greater topographic complexity. Numbers of live oysters and shell clusters were greater on the fore-reef and back-reef areas of both reef types. Second, laser transects were used to record reef profiles and the slope of fore-reef areas. Transect lines were stretched perpendicular to the water line and every 20 cm the distance between the lagoon bottom and reef top was measured. Vertical reef profiles and fore-reef slopes were significantly different between reference reefs and reefs with dead margins. Dead margins compressed reef widths, increased center peaks, and increased slopes on the fore-reef area by two-fold. Lastly, field experiments were conducted to determine the affect of dead margins on the vulnerability of oyster spat to predation. Structural variables (e.g. shell orientation, single versus shell clusters, reef slope) were manipulated and effects on oyster mortality were observed. Three predators were tested: the blue crab Callinectes sapidus, the common mud crab Panopeus herbstii, and the Atlantic oyster drill Urosalpinx cinerea. Structural variables did not have a significant influence on oyster mortality; however, a significant difference was established between predators. Panopeus herbstii consumed the most juvenile oysters, followed by U. cinerea and then C. sapidus. Together, these findings document ecological implications of dead margins on C. virginica reefs and reinforce the urgent need for enhanced regulations and restoration. If the intensity of recreational boating remains unregulated, dead margins will continue to increase. Thus, in order to maintain the diversity and productivity of Mosquito Lagoon, it is crucial to fully understand how dead margins alter the biogenic habitat and biotic communities of oyster reefs.