Robert M. Friedman, PhD - US grants

DESCRIPTION: (Adapted from the investigator's abstract) Lysyl oxidase (LO) functions as a suppressor of the tumorigenicity of the ras oncogene. In NIH 3T3 cells transformed by multiple copies of LTR-C-H-ras (RS485), the transcription of LO is markedly decreased. Long term treatment with interferon (INF) beta created revertants that still overexpressed ras but had restored LO expression. Transfection of persistent revertant cells with antisense lysly oxidase expression constructs led to retransformation and loss of LO expression. Because the ras oncogene is involved in several human cancers, it might be possible to prevent or treat such cancers by maintaining or restoring LO expression. Although LO is known as a secreted enzyme involved in extracellular collagen maturation, the gene is expressed in normal epithelial cells of breast, prostate, and colon. In human tumors derived from breast and prostate epithelium, LO expression is reduced or lacking. The mechanism(s) by which ras transformation down regulates LO expression will be studied. Preliminary studies of bisulfite modified genomic DNA suggest that the CpG island in the LO promoter is differentially methylated between NIH 3T3 and TS485. Methylation patterns in the LO promoter of NIH 3T3, RS485, and persistent revertant cells will be determined and compared to elucidate the possible contribution of methylation to the down regulation of LO expression. The mechanism by which restoration of LO expression suppressed the tumorigenic ras phenotype will also be investigated. In addition to its extracellular function in the maturation of collagen and elastin, LO was recently shown to be present and active in nuclei, suggesting that LO does have an intracellular function. LO deletion mutants will be used to determine which protein domains contribute to the functionality of reversion of FS485. Studies with antibody to IRF1 showed that in RS485 cells there was a protein smaller than IRF-1 that also bound IRF-1 antibody. The relationship of these two proteins and the contribution of the smaller species to LO transcription will be investigated. Treatment of RS485 cells with a combination of IFN beta and retinoic acid gave rise to a high percentage of revertants that had lost all of the multiple copies of the transforming LTR-c-H-ras oncogence. The mechanism involved in this deletion will be investigated as it might be useful for the treatment of HTLV or HIV induced diseases, which involve LTR-linked viruses.

Interferons (IFNs) have an unusual mechanism of action against some membrane-associated viruses. IFN-treated cells produce defective virus particles that have decreased infectivity because they are deficient in a membrane-associated glycoprotein. This is the principal antiviral activity of IFNs against retroviruses, but it is also an important mechanism for inhibiting the replication of vesicular stomatitis virus (VSV) in some cells such as IFN-treated LB cells which produce VSV particles of low infectivity that are deficient in VSV-G glycoprotein. In the IFN-treated cells G does not efficiently localize in the plasma membrane from which site it is ordinarily incorporated into budding VSV particles. Recent findings indicate that in IFN- treated cells the G protein of VSV is inefficiently transported to the plasma membrane from a cytoplasmic structure. Preliminary biochemical and morphological data suggest that this structure is the trans compartment of the Golgi complex (GC). The aims of this proposal are to analyze this localization in a clone of LB cells in which a large majority of the cells clearly manifest the cytoplasmic localization of G in IFN-treated cells. Experiments in IFN-treated cells will be carried out to study the morphological and biochemical localization of G. We also plan to analyze the transport of G protein within the GC. Other experiments will be focused on what biochemical effects of IFN might be involved in the inhibition of G transport such as changes in the pH of acid vesicles, in the fatty-acid composition of the cell, or in association of G with membranes. These studies may be useful in enlarging our understanding of how proteins are targeted by processing.

Leukotriene (LT) A4 hydrolase (E.C. 3.3.2.6) is a Zn-containing metalloenzyme classified as a cytosolic epoxide hydrolase. It catalyzes the conversion of LTA4 to LTB4, a potent pro-inflammatory agent. The crux of this investigation is to understand the nature and mechanism of small molecule inhibition of the hydrolase activity of the LTA4 enzyme with the objective to facilitate the design and development of pharmaceuticals to treat inflammation. Although aminopepidase activity was also found with this protein, its "natural" substrate has not been identified.

touch; somesthesis; sensory discrimination; brain electrical activity; somesthetic sensory cortex; proprioception /kinesthesia; behavioral /social science research tag; Macaca;

Description (provided by applicant): The goal of this proposal is to investigate how the brain encodes tactile motion information in the brain. Responses within different areas of primary somatosensory cortex (SI) will be examined (areas 3a, 3b, 1, and 2). Central to understanding their functional roles is finding out what distinguishes one area from another. By using simple versus complex stimuli, we aim to distinguish between areas whose responses are closely associated with the physical nature of the stimulus and areas whose responses are invariant and more closely reflect the motion percept. We will employ optical imaging, voltage sensitive dye imaging, BOLD fMRI, single unit recording, and anatomical tracing methods to address these questions. These experiments will elucidate the neural circuitries underlying tactile behavior and attention, understanding that will have clinical relevance for recovery of function from stroke and development of tactile prosthetics. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to investigate how the brain encodes tactile motion information in the brain, in particular in the primary somatosensory cortex (SI). These studies will elucidate the neural circuitries underlying tactile behavior and attention, understanding that will have clinical relevance for recovery of function from stroke and development of tactile prosthetics.

This unique history-of-science research project will investigate the Amundsen and Nansen polar explorations. Although the project coincides with the 100th anniversary of Amundsen?s South Pole expedition and the 150th anniversary of Nansen?s birth, the significance transcends the particularity of 2011. As part of the project, PI Robert Marc Friedman will develop a play focussing on the relationship between these two titans of polar exploration. An initial professional production of the proposed play is being planned, based on non-NSF funding. The dramatization will be based on contemporary scholarly research, as well as the research by the PI. Through this research and the development of the stage play the PI hopes to illuminate two primary themes: 1) the role of science in giving meaning and moral ballast to polar exploration, and 2) the role of native Arctic peoples? know-how as a critical resource for polar researchers in the age of heroic exploration.

The PI has several prior dramatizations about science, based on historical research, that have been professionally produced. Dr. Friedman wrote a play in 2002 about the process by which physicist Lise Meitner was denied a Nobel Prize, which was awarded to her male collaborator. Based on his experience with the Meitner stage drama, which has enjoyed numerous international performances and continues to be produced today also for conferences related to women in physics and the world nuclear-physics meeting, Dr. Friedman proposes that the Amundsen-Nansen play has similar potential to reach a broad audience, stimulate reflection, and create broader interest in polar science.

Although Amundsen and Nansen have previously appeared on screen and stage, this theatrical drama will bring the two men face to face for the first time. As Friedman states, "A stage play should not be a history lecture. While seeking to be faithful to the scholarly record, the play will nevertheless create its own theatrical reality and seek to grasp and hold the audience as a compelling drama. Why bring these two men together onto a stage? Why allow audiences to be drawn into an epic struggle of accusation, anger, envy, and admiration between two giants of polar exploration? How might we deconstruct myth and legend, expose the ignoble along with the noble, and yet find opportunity to celebrate?" These are questions that hold great interest not only to historians of science but anyone interested in the processes of science and how individual personality plays a role in the construction of scientific discovery itself.

In addition to the play and research the PI will create materials to accompany
performances into high school and university classrooms in order to expose students
to broader questions about the process of science, as well as create interest in
polar science.

The insights gained from the research entailed for the stageplay will be used as a starting point to analyze exchanges of knowledge about polar environment between members of what is often taken to be incommensurable cultures of knowing. Exchanges of knowledge between native Arctic peoples and professional scientists is increasingly receiving attention among Arctic social and natural scientists. Building upon a growing literature on this topic, the PI will develop research programs for studying such exchanges historically. Were those explorers who were willing to learn Arctic natives? techniques of travel, making clothing, building shelter, and preparing food also willing to consider these peoples? observations and insight about nature? What did explorers, including scientists, and Arctic peoples learn from each other about the nature of ice, aurora, weather and climate, behavior and migratory patterns of animals, or plant ecology?

? DESCRIPTION (provided by applicant): The goal of this proposal is to investigate the organization and circuitry within the hand and digit region of primary somatosensory cortex in nonhuman primates. Hand and digit behavior are defining aspects of object identification in primate behavior, and are central to different types of tool use, typing, texting, and body language. Despite this, little is known regarding the cortical circuitry underlying different stage of sensory (tactile and proprioceptive) processing. This proposal will use multiple anatomical and functional approaches to discover the intra-areal and inter-areal connectivity patterns between functional columns in the hand representation of primary somatosensory areas (3a, 3b, 1, and 2). These approaches will include anatomical tract tracing and functional tract tracing (using optical imaging and fMRI during focal electrical and pulsed near infrared laser stimulation). The role of these circuits during behavior will be examined. These studies will provide invaluable data for incorporation of sensory guidance in manual motor prosthetics, an area in which there is currently little understanding.

PROJECT SUMMARY Understanding how the brain processes what we see from the very center of our visual gaze (foveal vision) is essential for maintaining healthy vision, including high spatial acuity vision, color vision, and visual attention. Deficits of central vision lead to blindness as in the case of macular degeneration, or loss of depth perception as in the case of lazy eye (amblyopia). Macaque monkeys have visual systems very similar to humans and thus are an ideal animal model for studies of foveal vision. During the first few months of postnatal development in the macaque monkey and first few years of life in the human, retinal photoreceptors undergo an important migration to establish the adult foveal cone and rod distribution. Despite the importance of this part of vision, there is very little known regarding cortical representation of the fovea during development. Here, we investigate the relationship between postnatal retinal photoreceptor migration and changes in foveal cortical representation in infant monkeys. Using a combination of in vivo retinal photoreceptor imaging and cortical optical imaging and electrophysiological approaches, we aim to answer questions regarding the relationship between retinal cone photoreceptor migration and changes in foveal cortical representation in the first few months of postnatal development. Multiple developmental timepoints will be studied over the first 3 postnatal months, a period when foveal cone density can be mapped with adaptive optics. Paired retinal and cortical investigation will be conducted and data correlated. Revelations regarding the mechanisms of cortical plasticity during development will have great impact on understanding the development of central vision, computational models of cortical development, and on understanding retinal and cortical bases of visual developmental disorders which may lead to new approaches to treat neurological diseases like amblyopia and improve capabilities of brain-machine interfaces for the treatment of blindness.

PROJECT SUMMARY Decades of studies in animals have been conducted in conditions of strict experimental control of perception and behavior in order to understand the functions of the brain. For instance, in the study of visual perception visual paradigms often involve unnaturally long eye fixation periods and attention to locations away from the center of gaze (covert attention). These animal paradigms require long training periods (months to a year). While such studies have advanced our knowledge of brain function tremendously, a recurring question is how relevant the gathered data are to natural behavior. Today, the availability of cutting-edge technologies and computational power can free us from these limitations of the classical experimental paradigms and usher in a new generation of neuroscience questions focused on how the functioning of the brain in real world settings. The goal of this project is to develop a small wireless multimodal device for stimulating and recording domain- based cortical activity in freely moving primates. We will study the function of primary visual cortical areas with intrinsic optical imaging and optogenetic stimulation using a multimodal device that consists of a wireless camera and a wireless multisite LED stimulator. To evaluate the functionality of this approach, animals will perform visual detection and discrimination tasks. The end goal is the production of a multimodal device, constructed from off the shelf components, that provides unrestrained, multiareal and targeted (to specified cortical domains) stimulation and recording capabilities. The successful outcome of this project will contribute to our understanding of cortical encoding of perception and behavior and will have clinical relevance for the development of brain-machine interfaces.