Morris Benveniste - US grants

[unreadable] DESCRIPTION (provided by applicant): Temporal lobe epilepsy, the most common form of adult focal epilepsies, is often resistant to anticonvulsant therapy. Recently, surgery outcome has been improving, however, 15 - 30% of patients still report having seizures 5 years after surgery. Although it is generally agreed that epilepsy results from a disruption of the balance between excitation and inhibition, genetic and environmental factors and possibly a brain insult such as anoxia, infection or head injury may be involved in the expression of TIE. In many cases, the precipitating brain insult cannot be clinically recognized, and thus epileptogenesis may be difficult to detect and stop, prior to the development of the first spontaneous seizure. Thus, one line of treatment should involve cessation of spontaneous seizures once they occur. Anticonvulsants used to limit or prevent acute seizures might not prevent seizures in a chronic epilepsy. This indicates that mechanisms involved in generating acute seizures may be different from those involved in maintaining a chronic epileptic state. To develop drugs against a chronic epileptic state, the mechanisms involved in spontaneous seizures generation and maintenance after the latent phase should be elucidated. Synaptic activation of NMDA receptors has been reported to be increased after kindling. The high sensitivity of NMDA receptor activation to afferent firing rate raises the possibility that enhanced NMDA receptor responses to low frequency afferent firing in the epileptic state contributes to reduced seizure threshold. Increased expression of postsynaptic glutamate receptors would increase the degree of depolarization during repetitive stimulation. NMDA receptor activity in the hippocampus is altered after kindling, but the nature of the changes in NMDA receptors after epileptogenesis has not been elucidated. NMDA receptors bind glutamate with a higher affinity than AMPA receptors and lengthen the EPSP time course. Differences in biophysical properties of different receptor subtypes such as glutamate dissociation rates and magnesium binding affinities can also contribute to increased depolarization in the postsynaptic cell. We plan to determine if temporal summation of EPSPs is increased in CA1 pyramidal cells of the hippocampus in chronically epileptic rats, and if such increases are mediated by NMDA receptors. Seizures associated with Temporal Lobe Epilepsy are often generated in the hippocampus. This project focuses on determining how glutamate receptor mediated excitation changes in seizure prone CA1 pyramidal cells in the hippocampus. [unreadable] [unreadable] [unreadable]

2(3H)-Furanone, 3-ethyldihydro-4-((1-methyl-1H-imidazol-5-yl)methyl)-, (3S-cis)-; ARIA; Ammon Horn; Antibodies; Cell Death; Cell Nucleus; Common Rat Strains; Computer Programs; Computer software; Confocal Microscopy; Cornu Ammonis; DNA; Daily; Deoxyribonucleic Acid; Designing computer software; Diabetic Retinopathy; Electromagnetic, Laser; Ensure; Equipment; Experimental Designs; Fibroblasts; Fluorescence; Fluorescence Microscopy; Freeze Sectioning; Frozen Sections; GGF; Gel; Gene Products, RNA; Genetics, in situ Hybridization; Genotype; H and E; Harvest; Heart; Hematoxylin and Eosin; Hematoxylin and Eosin Staining Method; Hippocampus; Hippocampus (Brain); Histology; Human Resources; Hypothalamic structure; Hypothalamus; Image; Image Analyses; Image Analysis; Immunologic, Luciferase; In Situ Hybridization; In Situ Nick-End Labeling; In Vitro; Individual; Institutes; Investigators; Laboratories; Lasers; Light; Liver; Luciferases; Mammals, Mice; Mammals, Rats; Manpower; Maps; Methods; Methods and Techniques; Methods, Other; Mice; Mice, Transgenic; Microscope; Microscopy; Microscopy, Confocal; Microscopy, Fluorescence; Microscopy, Light, Fluorescence; Modeling; Molecular; Morbidity; Morbidity - disease rate; Morphology; Mortality; Mortality Vital Statistics; Murine; Mus; NDF; NRG Proteins; Neu-Differentiation Factor; Neuregulins; Neurosciences; Neurosciences, Other; Nucleus; Optics; Organ; PCR; Participant; Photoradiation; Pilocarpine; Polymerase Chain Reaction; Production; Programs (PT); Programs [Publication Type]; Protein Localization; Protein Trafficking; Publications; RNA; RNA, Non-Polyadenylated; ROC Analysis; RT-PCR; RTPCR; Radiation, Laser; Rat; Rattus; Research; Research Personnel; Researchers; Retinal; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleic Acid; SCHED; Sampling; Scanning; Schedule; Schools, Medical; Scientific Publication; Sensory-and-motor-derived factor; Services; Signal Pathway; Slice; Software; Software Design; Staining method; Stainings; Stains; Status Epilepticus; Status Epilepticus, Generalized; Structure; Structure of suprachiasmatic nucleus; Subcellular Protein Targeting; Subcellular Targeting, Physiological; Suprachiasmatic Nucleus; System; System, LOINC Axis 4; TUNEL; Techniques; Time; Tissue Sample; Training; Transcript; Transgenic Mice; UV laboratory microscope; Ultraviolet Microscopes; Week; body system, hepatic; brain tissue; computer program/software; cryostat; day; density; design; designing; experience; fluorescence imaging; fluorescence microscope; fluorescence/UV microscope; fluorescent microscope; hippocampal; hypothalamic; image evaluation; imaging; immunocytochemistry; in situ Hybridization Staining Method; in vivo; intracellular protein transport; laboratory fluorescence light microscope; medical schools; melanopsin; necrocytosis; novel; organ system, hepatic; personnel; programs; protein localization location; retina ischemia; retinal ischemia; reverse transcriptase PCR; suprachiasmatic nucleus; terminal nick end labeling

The major aim of the Imaging and Microscopy Core is to facilitate the research of investigators by providing access to imaging and microscopy equipment that might be expensive to procure and maintain in an individual investigator's laboratory not dedicated to these techniques. This allows investigators to broaden the techniques available for their research within the Neuroscience Institute by allowing the burden of expenses to be shared between individual investigators, the Neuroscience Institute and Morehouse School of Medicine. The imaging core can provide equipment for brightfield, fluorescence and confocal microscopy and image acquisition. The hardware and software are designed to allow production and analysis of images associated with in situ hybridization, immunocytochemistry, histology, as well as digitization and analysis of gels produced from molecular techniques such as Northerns, Westerns, Southerns, PCR and RT-PCR. Images produced are publication quality. Among the U54 participants, the Davidson laboratory will use the cryostat and microscope imaging equipment of the core to perform pathological analyses on tissue samples harvested from mice on shifting light schedules. The Paul laboratory will use the cryostat in the imaging core to section brain tissue for in situ hybridization and subsequent optical density analysis of autoradiograph images. In addition, the Paul laboratory will use confocal microscopy to assist in mapping expression of the GABAA subunit expression in the suprachiasmatic nucleus and other hypothalamic nuclei using novel GABAA subunit antibodies. The Fukuhara laboratory will use brightfield and fluorescence microscopy to examine whether Bmall-luciferase DNA is successfully delivered to fibroblasts utilizing in situ hybridization and image analysis software provided by the core. Core equipment can also be used, under proper guidance by other Neuroscience Institute investigators and other Morehouse School of Medicine personnel.

DESCRIPTION (provided by applicant): Older adults have difficulties with learning and memory acquisition. Severe impairment may be the basis for senile dementia. A vast accumulation of evidence strongly suggests that long term potentiation of synaptic function (LTP) may underlie learning and memory. Disruption of LTP in the hippocampus is well correlated to the inability to acquire some forms of temporal-spatial learning. While LTP is readily induced in young animals, it becomes increasingly difficult in older animals. This may well correlate with some learning deficits as animals age and may also reflect impairment in memory acquisition and maintenance in the elderly. Despite mild memory impairment in many elderly individuals, the ability to learn is not lost. This leaves open the possibility that other orms of plasticity are possible and may not diminish with age. Understanding the physiological basis for these forms of plasticity may result in developing different paradigms for more efficacious learning for the elderly, as well as targeting these forms of plasticity for pharmacological treatment. Although synaptic plasticity may strongly influence postsynaptic neuronal action potential firing, the ability for EPSPs of a certain strength to induce an action potential may als be modulated. This last type of modulation can be defined as E-S plasticity. We have made a novel discovery that E-S plasticity is independent of LTP, in that changes in EPSP strength are not correlated with changes in the E-S relationship generated for single postsynaptic neurons. Our preliminary evidence suggests that E-S plasticity remains robust while LTP wanes with age. Utilizing the in vitro slice preparation of the CA1 region of the hippocampus from different aged rodents, we will determine the defining characteristics of E-S plasticity and how it differs from LTP, with the overall goal of discerning different types of plasticity that still may be rapidly induced by stimulation as we age. To accomplish this we have delineated the study into three aims: 1) To determine how the relationship between LTP and E-S plasticity changes with age; 2) to determine if E-S plasticity abides by Hebbian criteria of cooperativity, associativity and input specificity; and 3) to elucidate differences in the molecular signaling pathway between E-S plasticity and LTP. Although testing behavioral learning paradigms is beyond the scope of this proposal, we can eventually use pharmacological conditions determined for induction and block of E-S plasticity in the absence of LTP to see which types of learning may be specifically related to E-S plasticity. This study is ideally suited for one of the aims of the National Institute of Agng of to study the continuum of cognitive aging across the lifespan.

Diversification of the workforce of the biomedical sciences is of high strategic importance to the United States. Diversity of the workforce has been shown to increase diversity of thought, improve creativity and hasten progress. In addition, with the increase in diversity within the population of the United States in the next 30 years, we will need a diversified workforce to keep up with the demand for jobs in the health and biomedical sciences. The BS/MS Program in Neuroscience of the Atlanta University Consortium has a long-term goal of increasing the proportion of and success of under-represented minorities in the field of Neuroscience. The underlying hypothesis of this grant application is that exposure of students early in their collegiate academic career to neuroscience research will lead to more of these students choosing a career related to neuroscience research. In addition, this exposure will fortify their background in neuroscience, and give them valuable experience in critical and quantitative thinking as well as experimental design. Under-represented minority students are recruited to the dual degree program near the end of their sophomore year. As Juniors and Seniors, students take a rigorous, graduate level neuroscience core curriculum at Morehouse School of Medicine, while still attending their undergraduate institution. The close proximity to Morehouse School of Medicine of the participating institutions of the Atlanta University Consortium (Morehouse College, Spelman College and Clark Atlanta University) allows students to walk to their graduate classes and laboratories on a daily basis. In this way, students can concurrently fulfill requirements for both the Bachelor of Science degree at their home institution and the master?s degree in Neuroscience granted by Morehouse School of Medicine. In the summer between Junior and Senior years, students start mentored laboratory work that contributes toward their master?s thesis. After graduating from their undergraduate institution, these students will continue to spend 10 ? 11 more months almost exclusively devoted to bench research. During the Master year, BS/MS students also participate in an intensive MATLAB bootcamp with beginning Ph.D. students in the Program in Neuroscience at Harvard Medical School. In addition to sharpening quantitative skills, the Harvard experience is a great networking opportunity. Career counseling of BS/MS students is given throughout the three-year program. The Master year ends with a public thesis defense leading to the earned master?s degree. In order to attract top tier students from the Atlanta University Consortium, tuition is paid by the program and students also receive a stipend for their time working in the laboratory. This experience will hopefully increase the probability of under-represented minority students to become successful in career tracts as Ph.D. scientists, M.D., Ph.D. physician scientists, or physicians interested in neurology, neurosurgery or other fields where they can participate in research or easily collaborate with researchers.