Andrew C. Bellemer, PhD - US grants

[unreadable] DESCRIPTION (provided by applicant): Galphao is the most abundant G protein in the mammalian brain and mediates neurotransmission by clinically relevent neurotransmitters such as serotonin, dopamine, and opioids. Despite its widespread role, the mechanisms by which Galphao signals is not known. C. elegans Galphao is more than 80% identical to the mammalian protein and acts to inhibit neurotransmitter release by an unknown mechanism. To identify this mechanism, a large-scale genetic screen was conducted for suppressors of an activated G protein-coupled receptor that signals constitutively through Galphao. The mutations isolated from this screen may identify molecules that function downstream of Galphao to inhibit neurotransmitter release. Aim 1 of this proposal is to map and clone the genes identified by the suppressor mutations. One gene has already been cloned, and four others have been mapped to small intervals. Aim 2 proposes using behavioral, cellular, and biochemical analyses to determine whether these genes encode downstream effectors of Galphao and to elucidate the mechanisms by which they inhibit neurotransmitter release. Thus the suppressors will be analyzed for defects in behaviors characteristic of defects in Galphao signaling, such as defects in egg- laying behavior and sensitivity to exogenous serotonin. Several of mutants have already shown such defects. Further experiments will determine in what neurons the genes are expressed and function, thus determining if the genes act in the same cells as Galphao. Aim 3 is based on the identification of one of the suppressor genes as encoding potassium-chloride co-transporter, KCC-3. This has led to the hypothesis that Galphao may inhibit neurotransmitter release by regulating chloride flux across the plasma membrane. This hypothesis will be tested through fluorescent imaging of chloride flux in intact worms using the genetically encoded chloride sensor, Clomeleon, and through genetic analysis of genes that encode proteins involved in chloride flux. The identification and analysis of the molecules that mediate the inhibition of neurotransmitter release by Galphao will advance the current understanding of Galphao signaling, as these mechanisms are currently entirely unknown, and may have broad implications for the understanding of basic nervous system function and the molecular phenomena underlying neuropsychiatric disorders. Public Health: The proposed research will investigate the mechanisms by which specialized signaling proteins known as G proteins function. Many neurotransmitters that are important for the regulation of mood, appetite, sleep, and addiction signal using these proteins. A better understanding of the mechanisms by which G proteins signal may provide a better understanding of how these processes are regulated. [unreadable] [unreadable] [unreadable]

An award is made to Appalachian State University (ASU) through the NSF-Major Research Instrumentation program to acquire a new laser scanning confocal microscope (LSM). A LSM is an essential tool for studying cell and gene function in animals, plants, and microorganisms. The LSM also improves the quality and expands the scope of the research, training and educational activities in many scientific disciplines, including biology and chemistry. The new LSM will be housed in the Dewel Microscopy Facility (DMF) (http://casmifa.appstate.edu/) within the College of Arts and Sciences. The DMF serves as the core, multi-user microscopy facility for all departments at ASU as well as surrounding Appalachian communities, including high school students in western North Carolina. Principle investigators that use the facility train undergraduate and master?s level graduate students to utilize this imaging technique in coursework as well as research environments. On a broader level, ASU and the DMF support several programs that train college and high school students from underrepresented groups pursuing degrees. These programs include Upward Bound that provides academic support to high school students in the Appalachian region, the Forensic Program in the Department of Chemistry, and the Duke TIP for gifted middle and high school students. This instrument will also be used for other outreach programs at ASU, such as the STEM education program and annual NC Science Festival. This project will significantly enhance research productivity and educational opportunities at ASU. It will also strengthen ongoing services to communities and collaborations with other universities in the region. Thus, the new LSM will become an essential tool for current and future scientists as well as engineers in all of western North Carolina.

The new instrumentation primarily serves users from the Departments of Biology and Chemistry at ASU for individual research, research training programs, and teaching. The five major research projects that use the LSM are multidisciplinary including investigation of the roles of microtubules in root development; study of the evolution of gene regulatory mechanisms using zebrafish as a model system; research on the morphology and function of sensory neurons in Drosophila melanogaster; exploration of the biophysical processes behind the transport of negatively charged molecules through major facilitator superfamily proteins overexpressed in mammalian cell lines; and examination of the evolutionary transitions in the mechanisms of ammonia excretion in vertebrates. The project award enhances these research areas as the LSM is configured with a pulsed 405nm laser for examining nuclear dyes, for photobleaching experiments and for laser ablation studies. Furthermore, additional lasers include a 25mW Argon with lines 458/488/514nm for many yellow or green dyes, a Diode Pumped Solid State Laser at 561nm for red dyes, and a HeNe 633nm for near infrared dyes. Research projects benefit from increased resolution, enhanced scanning speeds and the ability to conduct fluorescence resonance energy transfer experiments. The new LSM meets both the current and future needs for research and education across multiple disciplines at ASU and western North Carolina.

Appalachian State University (Appalachian) proposes to build on an existing Inclusive Excellence initiative at Appalachian that has produced significant gains in the recruitment and success of underrepresented minority students, but which has not yet successfully increased the recruitment and success of women and underrepresented racial and ethnic minority STEM faculty. The ADVANCE Adaptation project called ADVANCE APPALACHIAN will support efforts to: 1) incorporate ongoing institutional climate change into STEM departments by training of chairs and faculty using a framework of Inclusive Excellence; 2) support STEM faculty by developing new professional development and mentoring with attention to issues impacting equity for women and underrepresented racial and ethnic minority faculty; and 3) create new and improve existing work-life integration resources and advocacy across campus. Appalachian is a rural, masters level institution in North Carolina.

The goals of the project are based on analysis of institutional demographic and survey data and adapt strategies that have been demonstrated to be successful at other ADVANCE institutions. The data highlighted deficiencies in recruitment and retention of women and underrepresented racial and ethnic minorities into STEM faculty positions, as well as structural disconnects between senior university leadership and faculty that may limit the effectiveness of ongoing equity efforts. Analysis of successful ADVANCE projects submitted by other institutions showed that this barrier is not unique. Therefore, ADVANCE APPALACHIAN will implement activities such as training department and college leadership, providing professional development opportunities for all STEM faculty, and implementing work-life initiatives designed to target deficiencies in support for women in STEM at Appalachian and have the potential to produce sustained change that would reshape the environment for women in STEM at Appalachian. In addition, the ADVANCE APPALACHIAN project will form a partnership with the North Carolina American Council on Education (ACE) Women?s Network to expand the network of mentors and professional development opportunities for women STEM faculty at Appalachian.

The NSF ADVANCE program is designed to foster gender equity through a focus on the identification and elimination of organizational barriers that impede the full participation and advancement of diverse faculty in academic institutions. Organizational barriers that inhibit equity may exist in policies, processes, practices, and the organizational culture and climate. ADVANCE "Adaptation" awards provide support for the adaptation and adoption of evidence-based strategies to academic, non-profit institution of higher education as well as non-academic, non-profit organizations.

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.

While it is understood that students experience tremendous educational benefits from exposure to authentic research as part of their undergraduate coursework, the availability of these experiences may be limited by high cost and difficult implementation. The Connectomes for Undergraduate Neuroscience Education and Learning (CUNEL) project seeks to provide students with access to cutting-edge neuroscience research tools and provide the framework to integrate these tools into their undergraduate education. The CUNEL project will accomplish this by recruiting and training a network of neuroscience instructors who will implement and assess laboratory protocols that provide authentic research experiences in connectomics, the burgeoning field of neuroscience that seeks to create comprehensive maps of synaptic connectivity, the ‘wiring diagram’ showing how nerves connect within a nervous system. The broad outcome of the CUNEL project will be to enhance both the quality and accessibility of neuroscience education, while providing undergraduate students with real-world scientific experience within an area of neuroscience at the forefront of the field. <br/><br/>The CUNEL project will create, implement, and assess undergraduate laboratory materials that make use of the publicly available Female Adult Fly Brain (FAFB) dataset and Collaborative Annotation Toolkit for Massive Amounts of Image Data (CATMAID) software platform. The CUNEL team will create instructional materials that facilitate adoption, implementation, and assessment of FAFB/CATMAID laboratory modules in undergraduate classes at diverse undergraduate institutions. These modules will be disseminated and assessed through a user network of neuroscience instructors who will be recruited, supported, and coordinated by the project leadership. Students who work with laboratory materials created by the CUNEL project will have opportunities to engage scientifically with students at other network institutions and with the connectomics research community. The outcomes of this project will include: 1) the creation of laboratory materials that can be successfully implemented at a wide variety of undergraduate institutions to provide authentic research experiences in connectomics as part of undergraduate coursework, and 2) the creation and organization of a network of neuroscience educators who will collaborate to enhance neuroscience education using these authentic research tools.<br/><br/>This project is being jointly funded by the Directorate for Biological Sciences, Division of Biological Infrastructure, and the Directorate for Education and Human Resources, Division of Undergraduate Education as part of their efforts to address the challenges posed in Vision and Change in Undergraduate Biology Education: A Call to Action (http://visionandchange/finalreport/).<br/><br/>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.