FM1-43 dye ultrastructural localization in and release from frog motor nerve terminals.

Previous work has shown that the fluorescent styryl dye FM1-43 stains nerve terminals in an activity-dependent fashion. This dye appears to label the membranes of recycled synaptic vesicles by being trapped during endocytosis. Stained terminals can subsequently be destained by repeating nerve stimulation in the absence of dye; the destaining evidently reflects escape of dye into the bathing medium from membranes of exocytosing synaptic vesicles. In the present study we tested two key aspects of this interpretation of FM1-43 behavior, namely: (i) that the dye is localized in synaptic vesicles, and (ii) that it is actually released into the bathing medium during destaining. To accomplish this, we first photolyzed the internalized dye in the presence of diaminobenzidine. This created an electron-dense reaction product that could be visualized in the electron microscope. Reaction product was confined to synaptic vesicles, as predicted. Second, using spectrofluorometry, we quantified the release of dye liberated into the medium from tubocurarine-treated nerve-muscle preparations. Nerve stimulation increased the amount of FM1-43 released, and we estimate that normally a stained synaptic vesicle contains a few hundred molecules of the dye. The key to the successful detection of released FM1-43 was to add the micelle-forming detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), which increased FM1-43 quantum yield by more than two orders of magnitude.

Spatial learning with a minislab in the dorsal hippocampus.

We have determined the volume and location of hippocampal tissue required for normal acquisition of a spatial memory task. Ibotenic acid was used to make bilateral symmetric lesions of 20-100% of hippocampal volume. Even a small transverse block (minislab) of the hippocampus (down to 26% of the total) could support spatial learning in a water maze, provided it was at the septal (dorsal) pole of the hippocampus. Lesions of the septal pole, leaving 60% of the hippocampi intact, caused a learning deficit, although normal electrophysiological responses, synaptic plasticity, and preserved acetylcholinesterase staining argue for adequate function of the remaining tissue. Thus, with an otherwise normal brain, hippocampal-dependent spatial learning only requires a minislab of dorsal hippocampal tissue.

Limbic epilepsy in transgenic mice carrying a Ca2+/calmodulin-dependent kinase II alpha-subunit mutation.

Multifunctional Ca2+/calmodulin-dependent protein kinase II (CaMK) phosphorylates proteins pivotally involved in diverse neuronal processes and thereby coordinates cellular responses to external stimuli that regulate intracellular Ca2+ [Hanson, P. I. & Schulman, H. (1992) Annu. Rev. Biochem. 61, 559-664]. Despite extensive study, the impact of this enzyme on control of the excitability of neuron populations in the mammalian nervous system in situ is unknown. To address this question, we studied transgenic mice carrying a null mutation (-/-) for the alpha subunit of CaMK. In contrast to wild-type littermates, null mutants exhibit profound hyperexcitability, evident in epileptic seizures involving limbic structures including the hippocampus. No evidence of increased excitability was detected in mice carrying null mutations of the gamma isoform of protein kinase C, underscoring the specificity of the effect of CaMK. CaMK plays a powerful and previously underappreciated role in control of neuronal excitability in the mammalian nervous system. These insights have important implications for analyses of mechanisms of epilepsy and, perhaps, learning and memory.

Age-related learning deficits in transgenic mice expressing the 751-amino acid isoform of human beta-amyloid precursor protein.

The beta-amyloid precursor protein (beta-APP), from which the beta-A4 peptide is derived, is considered to be central to the pathogenesis of Alzheimer disease (AD). Transgenic mice expressing the 751-amino acid isoform of human beta-APP (beta-APP751) have been shown to develop early AD-like histopathology with diffuse deposits of beta-A4 and aberrant tau protein expression in the brain, particularly in the hippocampus, cortex, and amygdala. We now report that beta-APP751 transgenic mice exhibit age-dependent deficits in spatial learning in a water-maze task and in spontaneous alternation in a Y maze. These deficits were mild or absent in 6-month-old transgenic mice but were severe in 12-month-old transgenic mice compared to age-matched wild-type control mice. No other behavioral abnormalities were observed. These mice therefore model the progressive learning and memory impairment that is a cardinal feature of AD. These results provide evidence for a relationship between abnormal expression of beta-APP and cognitive impairments.

Feedback circuitry within a song-learning pathway.

The song system of birds consists of several neural pathways. One of these, the anterior forebrain pathway, is necessary for the acquisition but not for the production of learned song in zebra finches. It has been shown that the anterior forebrain pathway sequentially connects the following nuclei: the high vocal center, area X of lobus parolfactorius, the medial portion of the dorsolateral thalamic nucleus, the lateral magnocellular nucleus of anterior neostriatum (IMAN), and the robust nucleus of the archistriatum (RA). We now show in zebra finches (Taeniopygia guttata) that IMAN cells that project to RA also project to area X, forming a feedback loop within the anterior forebrain pathway. The axonal endings of the IMAN projection into area X form cohesive and distinct domains. Small injections of tracer in subregions of area X backfill a spatially restricted subset of cells in IMAN, that, in turn, send projections to RA that are arranged in horizontal layers, which may correspond to the functional representation of vocal tract muscles demonstrated by others. We infer from our data that there is a myotopic representation throughout the anterior forebrain pathway. In addition, we suggest that the parcellation of area X into smaller domains by the projection from IMAN highlights a functional architecture within X, which might correspond to units of motor control, to the representation of acoustic features of song, or both.

A role for Dynamin-dependent endocytosis during Drosophila gastrulation

Gastrulation, a process conserved among many higher organisms, is the directed migration of cells into layers that will establish various tissues targeted to become anatomical structures. This process is accomplished through another conserved morphogenetic event, known as cell intercalation. Early in development, this movement of cells within an organized tissue leads to unique cellular arrangements where neighboring cells contract their shared interfaces in order to meet at a shared vertex. In this thesis, I present work that demonstrates a requirement for Dynamin-dependent endocytosis during these contraction events. Using quantitative analysis, I have identified varied cell behaviors during experiments which knockdown the function of dynamin. In addition, I demonstrate the existence of an antagonistic relationship between Dynamin and the Myosin II motor protein. Lastly, localization and functional studies I performed for this work suggest a role for Sorting Nexin proteins during plasma membrane reorganization required for Dynamin-dependent endocytosis.

The Contribution of Structured Activity and Deliberate Play to the Development of Expert Perceptual and Decision-Making Skill

The developmental histories of 32 players in the Australian Football League (AFL), independently classified as either expert or less skilled in their perceptual and decision- making skills, were collected through a structured interview process and their year-on-year involvement in structured and deliberate play activities retrospectively determined. Despite being drawn from the same elite level of competition, the expert decision-makers differed from the less skilled in having accrued, during their developing years, more hours of experience in structured activities of all types, in structured activities in invasion-type sports, in invasion-type deliberate play, and in invasion activities from sports other than Australian football. Accumulated hours invested in invasion-type activities differentiated between the groups, suggesting that it is the amount of invasion-type activity that is experienced and not necessarily intent (skill development or fun) or specificity that facilitates the development of perceptual and decision-making expertise in this team sport.

An Interactive Model to Teach Motor Skills

There has been a tremendous increase in our knowledge of hum motor performance over the last few decades. Our theoretical understanding of how an individual learns to move is sophisticated and complex. It is difficult however to relate much of this information in practical terms to physical educators, coaches, and therapists concerned with the learning of motor skills (Shumway-Cook & Woolcott, 1995). Much of our knowledge stems from lab testing which often appears to bear little relation to real-life situations. This lack of ecological validity has slowed the flow of information from the theorists and researchers to the practitioners. This paper is concerned with taking some small aspects of motor learning theory, unifying them, and presenting them in a usable fashion. The intention is not to present a recipe for teaching motor skills, but to present a framework from which solutions can be found. If motor performance research has taught us anything, it is that every individual and situation presents unique challenges. By increasing our ability to conceptualize the learning situation we should be able to develop more flexible and adaptive responses to the challege of teaching motor skills. The model presented here allows a teacher, coach, or therapist to use readily available observations and known characteristics about a motor task and to conceptualize them in a manner which allows them to make appropriate teaching/learning decisions.

Learning from the Nordic welfare model: what and how? EU Centre Singapore Working Paper No. 16, September 2013

This paper posits that the Nordic countries were able to ensure good standards of equality for its citizens, while at the same time maintaining decent levels of economic growth. This can be attributed to the Nordic countries’ more holistic approach towards social spending and their focus on uplifting the skill levels of its workforce. Thus, the notion that there must be a trade-off between economic performance and a more aggressive welfare regime should be examined more thoroughly. The debate for policy makers should perhaps be framed with regard to where the balance should be between growth and equity rather than a trade-off. Firstly, the paper will elaborate on what exactly the “Nordic model” is, based on a broad literature review. Next, the paper will unpack the key characteristics of the Nordic model and analyse if indeed expansive welfare provided through state support erodes work ethic and impact the economic competitiveness of countries. Next, the paper will provide an explanation for how the balance between economic and social objectives is maintained, in some of the Nordic countries. Lastly, the paper discusses whether the same balance can be achieved in Singapore.

Sistema de recomendação inteligente para uma plataforma de e-Learning

As plataformas de e-Learning são cada vez mais utilizadas na educação à distância, facto que se encontra diretamente relacionado com a possibilidade de proporcionarem aos seus alunos a valência de poderem assistir a cursos em qualquer lugar. Dentro do âmbito das plataformas de e-Learning encontra-se um grupo especialmente interessante: as plataformas adaptativas, que tendem a substituir o professor (presencial) através de interatividade, variabilidade de conteúdos, automatização e capacidade para resolução de problemas e simulação de comportamentos educacionais. O projeto ADAPT (plataforma adaptativa de e-Learning) consiste na criação de uma destas plataformas, implementando tutoria inteligente, resolução de problemas com base em experiências passadas, algoritmos genéticos e link-mining. É na área de link-mining que surge o desenvolvimento desta dissertação que documenta o desenvolvimento de quatro módulos distintos: O primeiro módulo consiste num motor de busca para sugestão de conteúdos alternativos; o segundo módulo consiste na identificação de mudanças de estilo de aprendizagem; o terceiro módulo consiste numa plataforma de análise de dados que implementa várias técnicas de data mining e estatística para fornecer aos professores/tutores informações importantes que não seriam visíveis sem recurso a este tipo de técnicas; por fim, o último módulo consiste num sistema de recomendações que sugere aos alunos os artigos mais adequados com base nas consultas de alunos com perfis semelhantes. Esta tese documenta o desenvolvimento dos vários protótipos para cada um destes módulos. Os testes efetuados para cada módulo mostram que as metodologias utilizadas são válidas e viáveis.

Definition and diagnostic criteria of sleep-related hypermotor epilepsy

The syndrome known as nocturnal frontal lobe epilepsy is recognized worldwide and has been studied in a wide range of clinical and scientific settings (epilepsy, sleep medicine, neurosurgery, pediatric neurology, epidemiology, genetics). Though uncommon, it is of considerable interest to practicing neurologists because of complexity in differential diagnosis from more common, benign sleep disorders such as parasomnias, or other disorders like psychogenic nonepileptic seizures. Moreover, misdiagnosis can have substantial adverse consequences on patients' lives. At present, there is no consensus definition of this disorder and disagreement persists about its core electroclinical features and the spectrum of etiologies involved. To improve the definition of the disorder and establish diagnostic criteria with levels of certainty, a consensus conference using formal recommended methodology was held in Bologna in September 2014. It was recommended that the name be changed to sleep-related hypermotor epilepsy (SHE), reflecting evidence that the attacks are associated with sleep rather than time of day, the seizures may arise from extrafrontal sites, and the motor aspects of the seizures are characteristic. The etiology may be genetic or due to structural pathology, but in most cases remains unknown. Diagnostic criteria were developed with 3 levels of certainty: witnessed (possible) SHE, video-documented (clinical) SHE, and video-EEG-documented (confirmed) SHE. The main research gaps involve epidemiology, pathophysiology, treatment, and prognosis.

Comparative data and analysis in state motor vehicle administration. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Peripheral vision training for motor vehicle drivers. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Toward Never-Ending Object Learning for Robots

Thesis (Ph.D.)--University of Washington, 2016-06

The effects of unconditional stimulus valence and conditioning paradigm on verbal, skeleto-motor, and autonomic indices of human Pavlovian conditioning

The effects of unconditional stimulus (US) valence (aversive electro-tactile stimulus vs. nonaversive imperative stimulus of a RT task) and conditioning paradigm (delay vs. trace) on affective learning as indexed by verbal ratings of conditional stimulus (CS) pleasantness and blink startle modulation and on relational learning as indexed by electrodermal responses were investigated. Affective learning was not affected by the conditioning paradigm; however, electrodermal responses and blink latency shortening indicated delayed learning in the trace procedure. Changes in rated CS pleasantness were found with the aversive US, but not with the non-aversive US. Differential conditioning as indexed by electrodermal responses and startle modulation was found regardless of US valence. The finding of significant differential blink modulation and electrodermal responding in the absence of a change in rated CS pleasantness as a result of conditioning with a non-aversive US was replicated in a second experiment. These results seem to indicate that startle modulation during conditioning is mediated by the arousal level of the anticipated US, rather than by the valence of the CS. (C) 2002 Elsevier Science (USA). All rights reserved.