Anita K. McCauley - US grants

Awarding the 1998 Nobel Prize in Medicine to researchers investigating nitric oxide (NO) in the periphery demonstrates the importance of NO as a modulator of nervous system function. However, the understanding of NO in the CNS has lagged behind the extensive progress in peripheral structures. The objectives of this research proposal are to increase our understanding of the dynamic roles NO plays in the CNS throughout the lifespan and to characterize NO release in the thalamus. The temporally distinct patterns of NOS expression in the thalamus make it an ideal model system to study retrograde and anterograde release. During development, NO may be important in guiding retinogeniculate axon termination. NO association with the parabrachial brainstem (PBR), a region involved in sleep/wake states, in the adult suggests it may be involved in modulating brain states, and may be a critical modulator of sensory information flow from the periphery to cortex. The aims of this proposal are 1) to determine the source of NO in the developing LGN, 2) to determine if retinogeniculate stimulation causes NO release in the developing LGN and 3) to determine if activation of the PBR pathway results in NO release in the adult thalamus. Using the ferret LGN as a model, immunocytochemistry will be used to identify the neuronal elements expressing NOS in Aim 1 while a slice preparation will be used to investigate NO release in Aim 2. The cat LGN will serve as a model for complementary in vitro and in vivo approaches for Aim 3. Terminals will be activated pharmacologically and electrically, and the continuous rate and level of NO release will be determined. The mechanism of NO release will be verified through manipulation of the bNOS system The specific aims and experiments in this proposal are the cornerstone of my dissertation project. At the time of funding, I will have completed my preliminary exams and be ready to devote 100 percent effort to this research proposal.

This award supports purchase of a research-grade, epifluorescence stereomicroscope together with a digital camera, image analysis system and a vibration-isolation table. The equipment will allow users to view and capture images of large, intact structures with the magnification permitted by a stereomicroscope, and to combine fluorescence with low magnification imaging. The equipment will be housed and maintained in the Microscopy Core Facility, where it will be accessible to all members of the Biology department and to members of other departments. The equipment will be used in diverse research programs that span four major focus groupings: Ecology, Evolution, and Systematics; Comparative Physiology and Behavior; Cellular and Molecular Biology; and Integrative Plant Biology. Within these broad groupings, faculty pursue research at the level of the intact organism by examining the structure and function of intact, often living organisms. The research uses include, among others, the study of in vivo responses to changing physiological conditions, the time-lapse imaging of animal behavior, and determination of gene expression and protein localization at the level of the whole organism. In addition to its role in research, the stereoscopic imaging system imaging will be used to expand the types of microscopy used in the training of significant numbers of undergraduate and graduate students, including use in formal coursework and individual projects.

An award has been made to Wake Forest University under the direction of Dr. Anita K. McCauley for the acquisition of a laser scanning confocal microscope that will be used in research and education in biology and physics. The instrument will allow fluorescent imaging in thick tissues in biological samples, and of nanoparticles in various studies in physics. This provides a more three-dimensional view of the samples than available with conventional microscopes. Research projects that will be enhanced include studies of gene regulation of plant proteins, gene expression in bee brains, cellular behavior in insects, and excitable nanostructures in thin films. The institution will establish collaborations with several nearby colleges, including a historically Black college.

This Major Research Instrumentation award funds the acquisition of system upgrades for a Zeiss LSM 710 confocal microscope at Wake Forest University. The instrumentation enables faculty, postdoctoral fellows, graduate students, and undergraduates to study and publish important findings on brain plasticity; hormonal regulation of plant development; stress response systems, gene expression, and protein localization in Drosophila; mechanisms of DNA repair, protein complex formation, protein oxidation, and posttranslational modification. The requested accessories permit live-cell, dynamic, and multi-label experiments and enable more educational opportunities. Consistent with WFU's mission to strengthen connections beyond campus, the LSCM is also used by regional investigators for research and education, and is utilized during outreach workshops. The results of these research and teaching efforts will be broadly disseminated through abstracts and peer reviewed publications, as well as by active participation of students and faculty at professional meetings.

The primary research objective of this grant is to advance the fundamental understanding of the different physical and mechanical properties of cells as they advance through different stages of neoplastic transformation from normal to the metastatic state. Since recent reports indicate there is significant ambiguity about how these properties change for different cancer cells, the investigators plan to measure these properties for a single line of cells, and to determine whether the changes vary for different cellular components: i.e. whether the change in physical properties is due to a change in the cytoskeleton, the cell membrane, the cytoplasm, or a combination of these elements. Measurements using a wide array of techniques from physics and cellular biology will be applied to the different cellular structures. In addition, the investigators will create a biophysical computer model that brings together all measurements to account for the observed variations in physical properties at each state of neoplastic transformation.

This work will help to establish and disseminate new protocols and techniques for determining the differences in physical properties between cancerous and noncancerous cells. A team of physicists and biologists will train graduate and undergraduate students to work in this highly interdisciplinary field. Particular emphasis will be given to the education of minorities, through the connections to the MARC U* STAR programs of NC A&T University and Winston-Salem State University.