This masters thesis examines the hemispheric activation pattern of the cognitive processes involved in a complex mental rotations test (MRT) (Vandenberg and Kuse, 1978) using Steady-State Probe Topography (SSPT) (Silberstein et al, 1990) as a method to index brain activity. The Steady State Visually Evoked Potential (SSVEP) was recorded from 64 electrode sites using a multichannel electrode helmet, and elicited by a 13 Hz sinusoidal visual flicker, whilst the subjects were performing a visual vigilance Baseline task and the MRT. Forty-one right handed subjects (twenty male and twenty-one female) were used. In the MRT the subjects were required to choose the two figures which correctly matched the criterion figure in the centre. The figures were three-dimensional objects represented in two-dimensions on a computer screen. A significant finding of this study was that when all the subjects were considered as one group, no noticeable lateralization in cerebral activation associated with mental rotation was evident. When analyzing the results for the subjects, partitioned into two groups according to gender, evidence was found suggesting that the cortical processing associated with mental rotation may be more localized bilaterally in the males than the females. However, no noticeable lateralization effects for mental rotation were found in the males or females, and hence no gender differences in hemispheric lateralization was evident. An important finding was the emergence of gender differences in hemispheric lateralization in subsets of subjects performing with higher spatial ability. A left hemisphere lateralization for mental rotation was associated with the Best Performance Male group. The Best Performance Female group showed the opposite effect, where a right hemisphere lateralization was associated with better performance on the task. The lateralization effect appeared to be stronger in the Best Performance Males than the Best Performance Females. An important conclusion from this study is that when examining for hemispheric lateralization effects in mental rotation, and possibly other visual-spatial tasks, not only gender effects need to be considered, but the level of spatial ability in the comparison groups needs also to be taken into account.

The aim of this thesis was to investigate neuronal cellular mechanisms that may underlie the behavioural characteristics of the spontaneously hypertensive rat strain (SHR). The SHR was developed by selective breeding for elevated blood pressure and is also described as having increased levels of locomotor behaviour compared to its normotensive control strain, the Wistar-Kyoto. This hyperactivity and other behaviours, including altered sensitivity to reinforcement, have been used to model aspects of behaviour displayed in attention deficit hyperactivity disorder. In vivo intracellular recording of striatal spiny projection neuron activity in urethaneanaesthetised animals from three genetically related strains: the SHR, Wistar-Kyoto and standard Wistar, was employed to measure basic cellular properties and cellular mechanisms of reward-related learning. This population of neurons was chosen because alterations in their activity can influence behaviour and they are known to show cellular changes (synaptic plasticity) that are associated with learning. Cellular properties were measured in 71 neurons. Comparison between strains revealed a significant difference in action potential amplitude and duration between the SHR and Wistar-Kyoto strains. Interestingly, when measured at a later time, in a different sample of rats, the SUR action potential amplitude and duration were significantly different from the earlier sample. A change in the membrane potential repolarisation rate following action potential firing also occurred over this time. Twenty-nine of these neurons were also used in a study investigating the neuronal responses to a low dose of amphetamine (0.5 mg/kg). Changes were observed in some cellular properties following intraperitoneal administration of amphetamine. Synaptic plasticity at the corticostriatal synapses is sensitive to the timing of dopamine release in relation to cortical input. In anaesthetised preparations the spiny projection neuron membrane potential fluctuates between hyperpolarised (DOWN) and depolarised (UP) states, which reflect the level of cortical input. During the present study the responses of nine neurons to the induction of cortical spreading depression were observed to investigate the suitability of this method for use during synaptic plasticity experiments. Spiny projection neurons showed unpredictable responses to cortical spreading depression, therefore this method was not used further. Corticostriatal synaptic plasticity was induced in sixteen spiny projection neurons from two strains: SHR and Wistar. High frequency stimulation of the dopamine neurons in the substantia nigra, during the DOWN-state, did not induce any significant changes in corticostriatal synaptic efficacy. This was also true when high frequency stimulation of dopamine neurons was applied during the UP-state in neurons from the SHR strain. This thesis represents the first in vivo intracellular study of neuronal physiology in the SHR and Wistar-Kyoto rat strains. Results revealed action potential differences between these two behaviourally distinct rat strains. Synaptic mechanisms thought to underlie reward-related learning were not different between the SHR and Wistar strains, although the observed levels of plasticity were inconsistent with previous literature.

Lawrence S. Frishkopf. / "March 1, 1956." "This work was presented as a thesis submitted to the Department of Physics, M.I.T., 1956, in partial fulfillment of the requirements for the degree of Doctor of Philosophy." / Bibliography: p. 71-74. / Army Signal Corps Contract DA36-039 sc-42607 Project 102B Dept. of the Army Project 3-99-10-022

Klaus Putter. / "January 13, 1954." This report is identical with a thesis submitted to the Department of Electrical Engineering ... for the degree of Master of Science. / Bibliography: p. 33. / Army Signal Corps Contract DA36-039 sc-100 Project 8-102B-0 Dept. of the Army Project 3-99-10-022

Mammalian odorant receptors (ORs) are Class I G-protein coupled receptors (GPCRs) located within the nasal epithelium. Odorant receptors interact with Galpha olfactory, a Galpha S type G-protein. Activated Galpha olfactory stimulates adenylate cyclase and the resulting increase in cAMP concentration opens cyclic nucleotide gated channels allowing Ca2+ to enter the cell. The increased Ca2+ then activates a Ca2+ activated Cl- channel which further depolarizes the cell. This depolarization initiates an action potential that reaches the axon of the olfactory sensory neuron located in the main olfactory bulb. Information from the main olfactory bulb is then transmitted to higher regions of the brain. Olfactory information is initially coded through the interaction of odorant molecules with hundreds of distinct ORs, but difficulty in exogenous expression of odorant receptors has delayed the identification of ligands for individual ORs. However, expression of mouse odorant receptors in Xenopus laevis oocytes allows for a systematic screening for potential ligands, as well as for efficient study of the structure-function relationship of the receptors and their ligands. My screening of odorant receptors using Xenopus oocytes included the coexpression of a signal transduction system and the use of robotic two-electrode voltage clamp electrophysiology. In this study, I investigated the structural basis for ligand recognition in mouse odorant receptors. First, I expanded the molecular receptor ranges of seven Class I odorant receptors. By use of a high throughput assay, I was able to expand upon current knowledge in the field for the mouse odorant receptors 23-1, 31-4, 32-11, 40-4, 42-1, 42-2 and 42-3. I then examined one receptor (MOR23-1) in more detail. I used the substituted cysteine accessibility method to identify residues within transmembrane domain five of this receptor that are accessible from the extracellular space. These residues may line the ligand binding site or the ligand access pathway. Conventional mutations of A205 caused little alteration in the molecular receptive range of the receptor, suggesting that this residue may not play a significant role in ligand interaction within the binding pocket. Mutagenesis of G111, a residue within transmembrane domain three caused significant shifts in the molecular receptive range of the receptor, but the location of this residue within the binding pocket could not be confirmed by the substituted cysteine method. Previous reports had suggested significant similarity between the molecular receptive ranges of the seven mouse odorant receptors that I used in my research. By expanding upon the known aliphatic ligands for each receptor identified new ligands for each receptor, I was able to show that the molecular receptive ranges of these receptors are in fact distinct. The experimental identification of residues located within the binding pocket on transmembrane five of mouse odorant receptor 23-1 provides an improved understanding of ligand recognition by this receptor class and will aid in better computer modeling of these receptors. This increased accuracy of the computer models of these basic Class I GPCRs may aid in future drug discoveries. Since GPCRs constitute a significant fraction of current drug targets, understanding the mechanism of ligand interactions with mouse odorant receptors may aid in the development of more efficacious compounds in the treatment of many common ailments.

D-Aspartate (D-Asp) is an endogenous compound found in the central nervous system (CNS) of a variety of organisms. Despite its prevalence, however, relatively little understood of its physiological role. The prevailing theory is that D-Asp is an alternate agonist of N-methyl-D-aspartate receptor (NMDAR) channels. The goal of this work was to characterize the currents activated by D-Asp in neurons Aplysia californica, focusing on cells of the buccal S cluster (BSC). First, a general electrophysiological characterization was carried out, examining ion permeability, agonist dose-response, and the kinetics of activation, inactivation, and desensitization. D-Asp activated non-specific cation currents characterized by permeability to Na+ and K+. D-Asp-induced currents shared similar current-voltage relationships and time courses of activation and inactivation with L-glutamate (L-Glu)-induced currents. D-Asp currents, however, were subject to prolonged desensitization. Additionally, D-Asp activated currents independently of L-Glu, the known agonist of NMDAR channels, suggesting a non-NMDAR-dependent role of D-Asp. Next, select antagonists were used in an effort to pharmacologically characterize D-Asp receptor channels. These experiments suggested that D-Asp whole cell currents may be characterized by activation of multiple receptor sites, including NMDARS, excitatory amino acid transporters (EAATs), and a putative non-L-Glu D-Asp receptor. Furthermore, bath-applied D-Asp attenuated L-Glu-activated currents. Finally, D-Asp currents were compared to those evoked by acetylcholine (ACh) and serotonin (5-HT) in BSC cells. Results suggested that D-Asp activated receptor channels independently of ACh and 5-HT. Ten minute bath application of 5-HT was found to potentiate D-Asp current responses, likely through activation of a protein kinase C (PKC)-dependent mechanism, suggesting that D-Asp induced currents may be subject to synaptic plasticity associated with learning. While the identity of the putative D-Asp receptor remains elusive, the current work has advanced our understanding of the role D-Asp may play in the nervous system. These results should provide the groundwork for future studies aimed at identifying this unknown receptor channel, as well as investigation of the potential relationship of D-Asp receptor modulation to learning and memory in Aplysia, which may have relevance in higher organisms.

Carisbamate (CRS) and lamotrigine (LTG) are anticonvulsants which act mainly on neuronal voltage-gated sodium channels, that have been shown to have antidepressant-like effects in animal models of depression. In vivo electrophysiological recordings were carried out following 2 and 14 days of CRS or LTG administration. Overall firing activity in the dorsal raphe, locus coeruleus and ventral tegmental area were decreased with CRS. Similarly, a decrease in the dorsal raphe was also observed with LTG. Despite these presynaptic decreases in firing activity, both anticonvulsants exhibited significant enhancement of serotonergic transmission in the hippocampus as demonstrated by increased tonic activation of postsynaptic 5-HT1A receptors. This may be attributed to the observed desensitization of the terminal 5-HT1B autoreceptors. This study suggests that the enhanced serotonergic effect may be associated with an antiglutamatergic effect, and may contribute to the antidepressant-like effect of CRS in the forced swim test and the antidepressant properties of LTG.

Chronic iron overload (CIO) in patients leads to a cardiomyopathy characterized by conduction defects, including bradyarrhythmias. Using a murine model of CIO, we explored the effects of iron loading on the electrophyisology of the heart. Telemetric heart rate was reduced in conscious CIO mice compared to controls. Similarly, heart rates were depressed in both isolated CIO hearts and CIO mice following autonomic blockade, suggesting an intrinsic impairment of the SA node (SAN). Indeed, spontaneous action potential frequency was reduced in CIO SAN myocytes. The depressed pacing rate in CIO SAN myocytes was linked to reduced L-type Ca2+ current (ICa,L) density and a rightward shift in ICa,L activation, suggesting a selective reduction in α1D-mediated ICa,L. Western blot analysis demonstrates that the α1D isoform was reduced by ~ 89% in CIO atrial tissue. Therefore, the conduction defects under conditions of CIO are due to reductions in Cav1.3 channel expression in atrial tissue.

Chronic iron overload (CIO) in patients leads to a cardiomyopathy characterized by conduction defects, including bradyarrhythmias. Using a murine model of CIO, we explored the effects of iron loading on the electrophyisology of the heart. Telemetric heart rate was reduced in conscious CIO mice compared to controls. Similarly, heart rates were depressed in both isolated CIO hearts and CIO mice following autonomic blockade, suggesting an intrinsic impairment of the SA node (SAN). Indeed, spontaneous action potential frequency was reduced in CIO SAN myocytes. The depressed pacing rate in CIO SAN myocytes was linked to reduced L-type Ca2+ current (ICa,L) density and a rightward shift in ICa,L activation, suggesting a selective reduction in α1D-mediated ICa,L. Western blot analysis demonstrates that the α1D isoform was reduced by ~ 89% in CIO atrial tissue. Therefore, the conduction defects under conditions of CIO are due to reductions in Cav1.3 channel expression in atrial tissue.

Despite having similar brains, related species can display divergent behaviors. Investigating the neural basis of such behavioral divergence can elucidate the neural mechanisms that allow behavioral change and identify neural mechanisms that influence the evolution of behavior. Fewer than three percent of Nudipleura (Mollusca, Opisthobranchia, Gastropoda) species have been documented to swim. However, Tritonia diomedea and Pleurobranchaea californica express analogous, independently evolved swim behaviors consisting of rhythmic, alternating dorsal and ventral flexions. The Tritonia and Pleurobranchaea swims are produced by central pattern generator (CPG) circuits containing homologous neurons named DSI and C2. Homologues of DSI have been identified throughout the Nudipleura, including in species that do not express a dorsal-ventral swim. It is unclear what neural mechanisms allow Tritonia and Pleurobranchaea to produce a rhythmic swim behavior using homologous neurons that are not rhythmic in the majority of Nudipleura species. Here, C2 homologues were also identified in species that do not express a dorsal-ventral swim. We found that certain electrophysiological properties of the DSI and C2 homologues were similar regardless of swim behavior. However, some synaptic connections differed in the non-dorsal-ventral swimming Hermissenda crassicornis compared to Tritonia and Pleurobranchaea. This suggests that particular CPG synaptic connections may play a role in dorsal-ventral swim expression. DSI modulates the strength of C2 synapses in Tritonia, and this serotonergic modulation appears to be necessary for Tritonia to swim. DSI modulation of C2 synapses was also found to be present in Pleurobranchaea. Moreover, serotonergic modulation was necessary for swimming in Pleurobranchaea. The extent of this neuromodulation also correlated with the swimming ability in individual Pleurobranchaea; as the modulatory effect increased, so too did the number of swim cycles produced. Conversely, DSI did not modulate the amplitude of C2 synapses in Hermissenda. This indicates that species differences in neuromodulation may account for the ability to produce a dorsal-ventral swim. The results indicate that differences in synaptic connections and neuromodulatory dynamics allow the expression of rhythmic swim behavior from homologous non-rhythmic components. Additionally, the results suggest that constraints on the nervous system may influence the neural mechanisms and behaviors that can evolve from homologous neural components.