Grant Abstracts 2008   Buck Institute Julie K. Andersen Novato, CA $1,500,000 2008 Scientists at the Buck Institute share a common goal: to understand aging. We have recently made key discoveries that point to a deep mechanistic relationship between disease and aging across diverse species and various age-related diseases. Our collective data suggest that, rather than viewing aging as a risk factor, it may be more accurately viewed as a principal causal factor of these disorders. We propose to test the hypothesis that age-related human disease is the result of segmentally-accelerated aging – in other words, that diseases of aging such as Alzheimer’s disease, prostate cancer and osteoporosis are part and parcel of the fundamental aging process itself, simply enhanced in the affected tissues. This is based in part on our recent discovery of an acceleration of the aging process in terms of oxidative modification to mitochondrial complex I (CI) in affected regions of the Parkinsonian versus the normal aging brain; selective inhibition of this enzyme complex has been long associated with Parkinson’s disease (PD). To test our hypothesis, we propose two experimental programs: (1) to undertake a functional analysis of mitochondrial post-translational modifications (PTMs) particularly in CI to determine which are critical for neuronal cell dysfunction, death and progression of PD; and (2) to test the generality of this mechanism by surveying Alzheimer’s disease models and post-mortem patient brain tissues for mitochondrial PTMs and their functional correlates particularly but not limited to mitochondrial complex IV whose inhibition has been suggested to be preferentially involved in this age-related disorder versus changes associated with normal aging.   J. David Gladstone Institutes Lennart Mucke San Francisco, CA $1,500,000 2008 The ability to control one’s movements is essential to life. Neural circuits involving the basal ganglia are critical for proper motor control, and disruption of these circuits leads to movement disorders such as Parkinson’s disease and Huntington’s disease. The striatum, which is the input nucleus of the basal ganglia, is a major site of activity-dependent plasticity in both health and disease. Because the striatum lies upstream of other basal ganglia nuclei, cellular and synaptic plasticity within this region alters the transfer of information throughout basal ganglia circuits. However, studies of the striatum have been hampered by difficulties identifying different types of cells during both in vitro and in vivo experiments. Here, we propose to utilize recently developed optical and genetic technologies to characterize the properties of striatal neurons and the neural circuits in which they are embedded. In vivo fiber optic technology will be used in conjunction with expression of channelrhodopsin and halorhodopsin to identify and directly drive neural activity in striatal neurons of awake behaving mice. These experiments will allow us to address how the rate and timing of activity in basal ganglia circuits are causally related to motor behavior. The ultimate goal is to develop a framework that will enable the rational design of novel therapies for devastating disorders affecting the striatum.   Northwestern University Teresa K. Woodruff Evanston, IL $1,600,000 2008 Little is known about the signaling networks that support the integration of the male and female germ cells into a new totipotent cell, the one-cell embryo. We propose that heretofore poorly understood inorganic signaling molecules initiate the massive changes in the physiology of a fertilized egg. Based on preliminary studies, the team hypothesizes that fluxes in zinc ions mediate the first definitive signal in embryonic development. This hypothesis will be tested by two approaches: one targets real time changes in the subcellular concentrations of free zinc and calcium in live cells and the other rigorously maps specific changes in the total zinc pools at the nanometer level. The mouse oocyte is an ideal model system to study this novel inorganic signaling pathway. It undergoes a clear developmental pattern of receptor-mediated events as it transitions from a dormant stage to a fully active state upon fertilization. Also, its large size facilitates spatial localization of key molecular players. New analytical tools will be developed to map the abundance of specific inorganic molecules and biological receptors.   Stanford University Karl Deisseroth Stanford, CA $1,500,000 2008 Excitable cells are the biological building blocks of brain, heart and muscle, and communicate and compute using tiny, transient electrical currents. Widespread throughout the body, these cells underlie remarkable behaviors, from orchestration of movement to high-level cognition. When they malfunction, these cells give rise to devastating diseases, ranging from heart failure to Parkinson’s disease to depression; however, the interventional tools currently available to deal with these conditions are exceedingly primitive, have severe side effects, and yield little or no understanding of healthy cells or of disease processes. New technology is required, powerful enough to address the high speed and structural complexity of these electrical tissues. This grant is for a four-year project to address this fundamental obstacle by developing and applying emerging optical technologies first developed at Stanford. This light-based bioengineering approach has the power to control cellular functioning in vivo with millisecond precision, and to control intracellular messengers in specific cell types, thereby opening new vistas of both investigation and healing. The project personnel will develop the science and technology of this approach and apply these tools for the first time to mammalian models of neurological, neuropsychiatric and cardiac disease, spanning the central, peripheral and autonomic nervous systems.   University of Colorado at Boulder Natalie G. Ahn Boulder, CO $1,200,000 2008 This proposal requests support to develop innovative research strategies in mass spectrometry in order to detect and characterize all proteins in a cellular “proteome” (i.e., all proteins present in a single cell type). Mass spectrometry is the most powerful tool for addressing this problem and worldwide efforts are underway to define the proteomes of organisms, tissues and fluids. However, such efforts are stymied by the inability to comprehensively observe all proteins in highly complex biological samples. The goal is to create innovative experimental and computational technologies that will enhance the accuracy and sensitivity of protein detection by mass spectrometry, and enable comprehensive protein identification. In order to expand the depth to which the proteome can be observed, funds will be used to purchase a high resolution mass spectrometry system with capabilities for electron transfer dissociation. With this instrument, new methods will be developed to overcome limitations in data collection and solve major hurdles in recovering information from large scale datasets. These will be applied to collaborative research efforts between eight laboratories, providing the possibility for unprecedented capabilities to answer questions that were previously impossible to address.