Amanda Crocker, PhD - US grants

[unreadable] DESCRIPTION (provided by applicant): Sleep is one of the few remaining physiological phenomena that have systemic effects for which underlying molecular mechanisms are not known. While many important theories have been raised about its restorative function and, in recent years, its role in memory consolidation, no theory fully explains what might trigger sleep or waking. The overall goal of this work is to determine how sleep is regulated on a cellular and molecular level. Recent efforts by different labs have opened up the field of Drosophila genetics allowing for detailed analysis of sleep. Similar to mammals, Drosophila have an increased arousal threshold during sleep, show stereotypic rest positions and show rebound sleep following a period of deprivation. But Drosophila offer distinct advantages over mammals in research. They have a short regeneration time, genetic tractability and spatial and temporal control over transgenic expression. By using Drosophila to study the anatomical areas, cell populations and cellular pathways involved, we have the unprecedented opportunity to understand how and why animals need to sleep. Preliminary data indicate that sleep regulation can be mapped to specific loci in the Drosophila brain, in particular to the mushroom body (MB). We propose to identify the neurochemical basis of this regulation and refine the anatomical subdivisions that regulate sleep within the mushroom body. We hypothesize that sleep is regulated by specific neurochemicals acting in distinct regions of the MB. In order to address this hypothesis we plan to identia mushroom body specific octopamine receptor, two dopamine receptors and the serotonin 5HT-1a receptor. We also plan to examine the effects of modulating inputs, including dopamine, serotonin, and octopamine into the mushroom body. These studies will identify specific cells and molecules that regulate sleep in Drosophila. It is expected that at least some of these underlying mechanisms will be conserved in mammals, and will provide insight into the regulation and perhaps the function, of sleep. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): The brain has the amazing capacity to form long-lasting memories, but the underlying neural mechanisms remain elusive. To address this issue, I will use the Drosophila olfactory system to examine the cellular mechanisms underlying the formation and maintenance of long-term odor- associated memories. Drosophila are capable of forming long-term memories (LTMs) with underlying molecular pathways and synaptic modifications similar to those seen in higher organisms. A large body of work has defined the Mushroom Body (MB) as the neural substrate for olfactory learning; these studies have also determined that genes involved in memory formation are expressed within this structure. The study of LTM in the Drosophila olfactory system is facilitated by the fact that only a small number of MB extrinsic neurons (MBEs) read out synaptic changes associated with learning in the MB. While there exists a wealth of knowledge on olfactory processing prior to reaching the MBEs, the MBEs themselves remain largely uncharacterized. Here, I address how odor-associative LTMs are encoded at identifiable synapses within MB circuits. This proposal makes use of the microcircuit of the ¿ lobe compartment of the MB, consisting of Kenyon Cell (KC) axons and MBE dendrites. I will employ multiple state-of-the-art methods, including in vivo electrophysiology from identified MBEs, single fly learning and memory assays, along with single-cell gene expression analysis, to achieve a holistic understanding of olfactory memory encoding. Studying odor processing and memory formation at cellular resolution is required for developing novel strategies for the treatment of cognitive and neurodegenerative disorders that impact memory formation or recall. Our studies will also shed new light on the complex processes the brain uses to encode sensory stimuli and modify behavior. Specifically, my proposal will address the following aims: SPECIFIC AIM 1) To determine how odors are represented among a population of MB extrinsic neurons. SPECIFIC AIM 2) To characterize the relationship between behavior and neural signatures of LTM within MB extrinsic neurons. SPECIFIC AIM 3) To identify gene expression changes underlying olfactory LTM.

This Major Research Instrumentation Grant award supports the acquisition of a high-performance computing (HPC) system, the first large-scale, interdisciplinary research cluster at Middlebury College, a research intensive undergraduate institution. Cutting-edge research in the natural and social sciences utilizes digital technology to model and evaluate important social, environmental and scientific problems. Sophisticated modeling techniques and expanding datasets require the processing capabilities of a multi-processor server with the capability to read and analyze datasets sized as large as several hundred gigabytes or low terabytes. High-performance computing systems offer computing power that is useable across any discipline with computer modeling or data analysis applications. The research to be supported by this equipment is varied and impactful, ranging from studies of the impact of trade and agricultural technology on food markets to genomic studies of variation in pain sensitivity. This proposal includes nine separate research projects slated to use the HPC system with many more in queue. Overall, this substantial increase in processing capability enhances the scope of existing research at Middlebury and makes possible new projects that would have otherwise been impossible with existing computing infrastructure. HPC also supports faculty-mentored research-training programs in the social and natural sciences, fulfilling a critical part of the college's teaching mission. Middlebury College takes special effort to recruit and support underrepresented minorities in the STEM and STEM-related social sciences. HPC increases student access to faculty-guided research in these fields. The research to be conducted focuses primarily on human behaviors with the potential to refine best practices in neurological and genomic studies.


Research projects will leverage the HPC resource across political science, economics, neuroscience, computer science and biology. One project examines heterogeneous causes for unemployment and their impact on labor market recovery rates, suggesting more targeted corrective measures for unemployment. A neuroscience study explores pain perception and stress responses, focusing on how animals learn in response to noxious stimuli and tying particular genes to variation in responses. Two other projects involve DNA sequencing. One offers new modeling techniques to improve genotype prediction from observed data; another examines toxic algae blooms and investigates viruses as bloom-mitigation mechanisms using genome sequencing techniques. The flexibility of HPC makes it a multi-disciplinary tool to enhance research across STEM-related social sciences and more traditional STEM fields.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

An award was made to Middlebury College to acquire a confocal microscope. This award provides the rural areas outside central Vermont with access to a key piece of modern research technology. Access to this instrument allows for enhanced education and research opportunities for students and faculty members at Middlebury College, as well as for colleagues from Castleton University and Norwich University. At Middlebury College, this microscope will be used in multiple courses across disciplines including biology, neuroscience, and physics, as well as in student-directed independent studies. This will expose almost a fourth of the entire student body to new, cutting-edge research technology. Within these disciplines, half of the students are women, nearly one fourth are of an ethnic minority, and over 15% are Pell grant recipients or first generation college students. This grant also supports a number of community outreach events and activities. Free and reduced lunch rates are as high as 56% in the communities surrounding Middlebury College, indicating substantial rural poverty. This poverty combined with low college-enrollment rates limits the access of many school-age children to high quality research and learning experiences. The confocal microscope made available by this award will enable Middlebury College to offer several new opportunities to educate and inspire local school-age children.

This award also enables Middlebury and other regional faculty to pursue novel avenues of research, which were previously hampered or impossible. This award supports faculty across disciplines pursuing a wide range of research and education. In physics, a confocal microscope will allow faculty and students to employ a novel absorption-based imaging technique using the photothermal effect for contrast. In neuroscience, on-site access to a confocal microscope will allow faculty and student researchers mentored by accomplished faculty to investigate the development of the nervous system through multi- channel, high-resolution fluorescent immunohistochemistry and 3D reconstructions. This award also supports the pursuit of questions pertaining to the cellular response in neurons and glia to mechanical stress. The confocal microscope will allow Biology faculty to pursue questions involved in the regulation of vesicle release in C. elegans, memory formation in rats, and oocyte development. This award will improve undergraduate participation in advanced research by supporting a microscope that can rapidly colocalize multiple fluorescent labels from in-vivo and in-vitro specimens while providing an accessible user-interface for student learning. This will support the grantees? mission to train the next generation of scientists and policy makers. This award enables novel research avenues pursued by undergraduate students and faculty, and results will be disseminated in peer-reviewed journals and at local, national, and international scientific conferences.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.