Arthur N. Popper - US grants

The number of sensory hair cells in the auditory endorgans of the mammalian ear are not known to change after birth. In contrast, sensory hair cells continue to be added for at least several years (if not throughout the life of the animal) in the otic endorgans that have been studied in elasmobranchs, bony fishes, and amphibians. Since the sensory hair cells of these species are morphologically and functionally similar to those found in birds and mammals, elasmobranchs, bony fishes or amphibians could potentially provide 'model' systems for the experimental investigation of hair cell development, maturation, and cell death. This study will focus on the sensory hair cells of the ears of fishes since earlier work has demonstrated extensive addition of sensory hair cells (average of more than 167 cells per day) for a long time post-embryonically. However, in order to more fully make use of the fish ear as a system for the study of hair cells production we need to generate base-line data on the patterns of development in post-embryonic animals, to compare development to embryonic fish and mammalian cells, and to determine whether cell replacement occurs in adult animals. The specific investigations we will conduct will include: (a) morphological changes in sensory hair cells as they mature in post-embryonic and embryonic animals; (b) the sites on the post-embryonic and embryonic sacculi where new sensory hair cells are added; and (c) determination of hair cell replacement in ears in which hair cells are no longer being added. These studies will ultimately lead to investigations of the anamniotic ear with in vitro studies wehre it will be possible to experimentally manipulate the hair cells environment in order to study aspects of its growth and the effect on growth of such things as ototoxic drugs. Investigations will involve labeling proliferating tissue with 3H thymidine and then doing autoradiography to determine sites of cell addition. Light and transmission electron microscopy will be used for evaluation of tissue. Other investigations will involve use of scanning electron microscopy to determine the numbers of sensory cells in various otic endorgans.

Investigations concern sound detection and processing by fishes. The auditory system in fishes has a number of homologues with the auditory system in other vertebrates. Consequently, understanding functional aspects of hearing in fishes provides insight into general questions of vertebrate auditory system structural and functional relationships. Experimental investigations include measurement of hearing capabilities using operant conditioning techniques to determine hearing sensitivity and other psychophysical functions. Physiological studies are directed at understanding processing in the teleost ear and the roles of more peripheral structures, such as the swimbladder and Weberian ossicles, in sound detection and processing. Ancillary investigations include TEM and SEM studies of the ultrastructure and gross morphology of the ear in order to gain better insight into the functional anatomy of the ear in fishes. Studies are being conducted comparatively in order to ascertain when there is a 'typical' teleost auditory system or whether there are multiple mechanisms of detection and processing in fishes.

This proposal aims to investigate the extent to which isolated teeth from sharks of the genus Carcharhinus exhibit species-specific traits. This data set, once established, will serve as an essential reference against which fossil teeth can be compared in the future.

The Fourth International Congress of Systematic and Evolutionary Biology will be held at the College Park Campus of the University of Maryland from June 30 to July 7, 1990. The theme of the Congress "The Unity of Systematic and Evolutionary Biology" will bring together an international assemblage of systematists, population biologists, and ecologists, as well as members of the "user community", for which systematics is a critical but undervalued resource (e.g., agroeconomists, government policy specialists, conservationists, and biotechnologists). The Congress will host sessions on a variety of topics, several of which will be devoted to the main themes of: 1) Evolution in Perspective: Biodiversity, Conservation, Biotechnology, and Global Change; 2) Evolutionary Mechanisms and Processes; and 3) Systematics and Phylogenetic Reconstructions. Ten scientific societies will also be sponsoring symposia at the Congress. Dr. Reaka's proposal to the National Science Foundation addresses the critical issue of human resources in systematic and evolutionary biology. NSF support for the Congress will allow graduate students, members of under-represented groups of U.S. scientists, and investigators from lesser-developed countries to attend this important meeting. This travel support will promote international collaborations and enhance the research careers of women and minority scientists in the U.S.

9310484 Popper This U.S.-Russian Workshop on Comparative Sensory Biology is organized by dr. Arthur N. Popper of the University of Maryland, College Park and Dr. Mikhail A. Ostrovsky of the Institute of Chemical Physics of the Russian Academy of Sciences and professor at Moscow State University. In past years, it has become apparent that different animal species have evolved the ability to extract a good deal of sensory information within their environment. While some of these senses, such as vision and hearing, are similar in aquatic and terrestrial animals, many aquatic animals have evolved special techniques for extracting and processing information. In addition to the more "traditional" senses, the lateral line and electroreception are found uniquely in aquatic species. To date, there has been only a single international meeting on sensory systems of aquatic animals and one on comparative sensory systems, in 1987. Due to the political climate, no Russians were included in the meeting. Only a few Russians were included in a more general 1982 meeting. At the same time, there has been very active research in Russia in various areas of comparative sensory biology and evolution of sensory systems. Thus, this workshop will enable U.S. and Russian scientist to benefit from the interchange of ideas and from potential bilateral collaborations. The workshop topics will be limited to three sensory systems: electroreception, mechanoreception (hearing and the lateral line), and vision. This research in zoology fulfills the program objectives of advancing science by enabling leading experts in the U.S. and Russia to combine complementary talents and pool research resources in areas of strong mutual interest and competence.

This proposal is a joint effort of the Departments of Biology and Psychology and the Neuroscience and Cognitive Science Graduate Program at the University of Maryland, College Park. We have at UMD an exceptional group of 10 faculty with a common research interest in comparative hearing and evolution of hearing. The breadth of experimental approaches and species studies, provides us with an opportunity to provide research training at the graduate and postdoctoral level that exists no where else. At the core of the research training program are 10 funded investigators with proven research and training records, most of whom are located in the same building. There is a strong history of research interaction among this group, and this was further facilitated by the initial five year funding of this training grant. Graduate and postdoctoral students (as well as undergraduates) take advantage of the interaction between faculty and research groups. The proposed program will provide support for four predoctoral and three postdoctoral fellows each year. Two separate but closely related issues are emphasized in our program. The first is that comparative auditory function and structure, and the second is the evolution of the auditory system. The major goal of our program is to produce scientists who have an appreciation for, and an understanding of, the diversity of vertebrate hearing mechanisms and the evolution of the auditory system. We anticipate that the individuals trained in our program will conduct full-time research on evolutionary and/or comparative questions in academic settings. At the very least, our training will ensure that they will be able to put their work into the appropriate content and help add to our growing understanding of these issues. Predoctoral and postdoctoral trainees are recruited nationally. Very importantly, UMD and our participating departments and colleges have strong records of recruiting, retaining, and retention of women and minorities. We continue to take advantage of these opportunities to continue to make recruitment and retention of women and minorities a very important aspect of our program.

Ten biologists from the Departments of Zoology and Psychology of the University of Maryland, College Park are requesting funds to purchase a laser-scanning confocal microscope and image analysis workstation to support and enhance their research. Drs. Mount, Payne, Popper and Rivas will be the primary users of the instrument. Within our multidisciplinary group, the uses of confocal microscopy include investigations of neuronal polarity in cerebellar granule neurons (Dr. Rivas, Co-PI on this grant), regeneration of sensory hair cells in the ear (Drs. Dooling and Popper), the role of calcium ions in phototransduction (Dr. Payne), the development of central auditory projections in the CNS (Drs. Brauth, Carr, and Hall), how the correct processing of RNA from protein-coding genes is specified in higher organisms (Dr. Mount), homology of invertebrate auditory neurons (Yager), and investigations of stream meiofauna (Dr. Palmer). These laboratories have a large number of postdoctoral, graduate and advanced undergraduate students who will use the confocal microscope during their research training. We are requesting a Bio-Rad MRC 1024 confocal imaging system coupled to a Nikon 300 inverted epi-fluorescence microscope, and a Silicon Graphics "Indy." image analysis workstation for quantitative image analysis and three-dimensional image reconstruction. The confocal system will include a krypton-argon laser (with emission lines at 488, 568, and 647 nm), triple dichroic beamsplitter, four detection channels (three PMT-based detector channels and one detector for transmitted light), and high resolution 24-bit acquisition software. This system will allow us to image up to three separate fluorescent channels for multi-label applications. An inverted configuration with DIC optics is needed so that the epi-fluorescence Nikon microscope can be used for cell cultures, and to provide ample access for micromanipulation, as is required by several of our users. Confocal micr oscopy will significantly enhance the research programs of each of the core investigators, and will allow them to pursue areas of research that have heretofore been extremely difficult or impossible to examine with conventional microscopy. In contrast to conventional fluorescence microscopy, in confocal microscopy out-of-focus structures do not contribute to background because they receive little or no illumination, and any fluorescent signal produced by them is rejected by the instrument. Unlike conventional microscopy, where one must physically cut a thick tissue into extremely thin sections for imaging, the extremely shallow depth-of-field of confocal microscopy allows the optical sectioning of either intact cells or thick specimens (such as embryos or brain slices). Optical sections taken at successive focal planes can be re-combined using computer-generated techniques to produce a highly informative three-dimensional reconstruction of an entire tissue or cellular structure. These characteristics make this technique essential for our multi-disciplinary studies, all of which require optical sectioning through thick biological specimens with elimination of out-of-focus fluorescence. Each of the investigators in this proposal will use the optical sectioning and quantitative capabilities of the confocal imaging system. For example, Dr. Rivas will use the confocal to study the development of fluorescently-labeled neurons that were implanted into the brain. Using conventional fluorescence microscopy, out-of-focus fluorescence from thousands of implanted neurons masks the structure of any particular cell. Indeed, individual neurons extend through many focal planes, allowing sharp focusing only on particular portions of the cell. Drs. Brauth, Carr, Dooling, Hall, Popper, and Yager face similar methodological problems as they attempt to study the complex structure of neurons in the auditory nervous system. Dr. Mount will use the confocal to make optical sections th rough thick Drosophila embryos, and Dr. Palmer will use the confocal to section through invertebrate whole mounts. Dr. Payne will extend our use of the confocal into the area of neuronal cell physiology. Dr. Payne will investigate a localized light-induced elevation of intracellular calcium that is co-incident with the electrical response of photoreceptors to light. Drs. Rivas and Yager will also use the confocal for live neuronal imaging, using the optical sectioning capabilities of the confocal to image fluorescently-labeled neurons in thick brain slices or embryos. Thus, in each area of our research, the confocal will greatly improve our ability to study correlated structure and function.

The proposed study will investigate changes in the hearing capabilities and ear in aging zebrafish (Danio rerio). This teleost species has become a useful and important vertebrate model system for genetic studies, and it is of special interest since many features of the zebrafish ear, including the structure and function of the sensory receptors, are homologous to those in mammals. Among the numerous known zebrafish mutants, over 50 affect the ear as their primary target, while others affect the ear secondarily. Many of these mutations resemble those found in other vertebrates. Zebrafish have the added advantages, compared to the most popular genetic model for hearing studies, mice. These include their: having large broods; reaching sexual maturity at a relatively young age; having clear embryos; and being low cost to obtain and maintain in the laboratory. This study will investigate specific changes in the aging nervous system of zebrafish, starting before the fish reach sexual maturity and continuing through sexual maturity and into old age. The results will provide baseline data for understanding changes in the auditory system of the aging zebrafish, and potentially provide a model system in which we can apply known genetic changes in the ear to better understand the ways in which the vertebrate ear develops and ages. The study will test whether auditory sensitivity of zebrafish remains stable from a young age until a period past sexual maturity, but then get poorer in older animals. It will also determine the pattern of sensory hair cell and eighth nerve proliferation in post-embryonic fish. Auditory sensitivity will be determined using a classical conditioning paradigm that has been successfully used for other species. Hair cell addition will be done using flourescent markers and confocal microscopy.

The ability to detect ultrasonic sounds (frequencies over 20 kHz) is not limited to mammals among the vertebrates. Instead, American shad, a clupeid fish (related to herring) can detect. The shad ear is clearly involved in ultrasound detection, but the mechanism of ultrasound is detection is not known. The shad ear has a number of characteristics that are unique among vertebrates. Moreover, there is evidence that the support structure of the sensory epithelial of the utricle, the primary auditory endorgan in shad, may be functionally analogous to the high frequency regions of the mammalian organ of Corti. The proposed work is to do a detailed physiological and ultrastructural analysis of the ear and eighth nerve in order to determine how ultrasound is detected. It will also quantify the ultrasound detection capabilities of American shad and other-related species in order to get a better understanding of the kinds of signals that can be detected. Physiological studies will examine the response characteristics of each inner ear otolithic endorgan, and determine the ear region(s) involved with ultrasound detection. Neural responses from these regions will be quantified. Using extracellular and intracellular techniques. Ultrastructural and morphological approaches will be used to understand the structure of the ear, and the mechanical adaptations for ultrasound detection. Psychophysical studies at the University of Maryland will examine hearing capabilities at sonic and ultrasonic ranges, and characterize the ability of these animals to detect signals in noise. These studies will also help elucidate the precise mechanisms of ultrasound detection. Behavioral studies at Mote Marine Laboratory (Sarasota, Florida) will help characterize response to echolocation sounds, and help determine which species are response to ultrasound. These investigations will provide an opportunity to examine ultrasound detection in a highly accessible ear. This system has the potential of serving as a model system where ultrasonic region of the ear is amenable to investigation. This system has the potential for contributing to our understanding of how hair cell systems, and their innervating neurons, are able to respond to ultrasonic stimuli. Moreover, this system could be useful in analysis of how otolithic organs of the ear in mammals can potential contribute to hearing.

Technical Abstract

The University of Maryland will form a regional microscopy facility through the acquisition of key instrumentation for a pair of new transmission electron microscopes (TEMs). The requested equipment includes analytical spectrometers for a JEOL 2100F field-emission TEM (installation 2/06) and specialized specimen holders for use with a new JEOL 2100 LaB6 TEM (installation 7/06). The spectrometers include an energy dispersive X-ray spectrometer and an electron energy loss spectrometer with spectral camera. The specimen holders will allow heating to 1273 K and cryogenic specimen transfer at 77 K. These additions to our facility will create a one-of-a-kind regional university facility in the Washington, DC metropolitan area, and will be a shared experimental facility for the NSF-MRSEC at UMD. Some of the research areas to be addressed with the requested instrumentation include: (a) structure and composition of multi-functional materials (both hybrid organic-inorganic systems and multiferroic self-assembled nanocomposite films); (b) the understanding of auditory sensory cells in birds and fish; (c) the structure and composition of doped semiconductor nanomaterials for spintronic applications; and (d) phase transitions of ferromagnetic shape memory alloys. The microscopy facility will be used in educational programs for undergraduate, graduate and postdoctoral students, and for training student researchers at the University of Maryland and five university partners in the DC area. The microscopes will enhance our K-12 educational outreach activities by providing virtual microscopy courses over the web and support other campus activities such as the ASM Materials Training Camp for teachers and Project Lead the Way.

Lay Abstract

Transmission electron microscopy is one of the most powerful tools for research in the fields of nano- and bio- technology. These microscopes can magnify up to 1.5 million times and allow the observation of materials at the atomic level. Through this proposal, the University of Maryland will acquire spectrometers for our new microscopes allowing us to form a unique new regional facility for cutting edge electron microscopy research and training across all educational levels. The facility will service the needs of institutions throughout the Washington, DC metropolitan area, including five partner Universities. This facility builds on a strong foundation at the University of Maryland in the areas of electron microscopy, nanotechnology and bioscience and will allow researchers to find pathways to better magnetic sensors, faster microelectronic devices, materials that change shape under applied fields, new nanoparticle catalysts, the understanding of sensory cells used in hearing and other biological functions. Many of these projects are also keystone areas in the Materials Research Science and Engineering Center already funded at UMD by NSF. The microscopy facility will be used in educational programs for undergraduate, graduate and postdoctoral students, and for training student researchers at the University of Maryland and five university partners in the DC area. The microscopes will enhance our K-12 educational outreach activities by providing virtual microscopy courses over the web and support other campus activities such as the ASM Materials Training Camp for teachers and Project Lead the Way.

This award is made through the Ethics Education in Science and Engineering solicitation (NSF 06-524). The project will develop a set of ethics courses and workshops that focus on graduate-student education, but also provide training on research integrity for postdoctoral associates and for new assistant professors that will enable these individuals to be better "role models" in research ethics for their graduate students. These courses will be developed first within the colleges of the Principal Investigators at the University of Maryland, College Park, using the extensive experience of the Principal Investigators in such courses. In the second year of the grant, the courses will be extended to the other science and engineering colleges and departments on the College Park campus. In the third year, the courses will be made available across the State of Maryland in the University System of Maryland. The courses will engage graduate students in actively solving real-world research-integrity problems, and the influence of the courses will be extended by training postdoctoral associates as ethics teachers. These trained teachers will later disperse across this country and abroad. The course for graduate students, Research Ethics, will include an introduction to the philosophy of ethics and value systems, and an introduction to the philosophy of science and how science is structured. Then it will discuss the issues that arise in trying to do "good Science" and to avoid bias in research, and the cross-checks against error and bias. Other topics will include animal subjects, human subjects, attribution and authorship, mentoring, intellectual property, and under-represented groups. A second course, Advanced Research Ethics, will be developed for postdoctoral associates and for graduate students who wish to obtain the Graduate Certificate in Research Ethics that will also be developed. This course will develop the course topics more deeply and will cover the pedagogy of ethics training. A new Research Ethics Workshop will be developed as a 4- to 6-hour one-day workshop for untenured faculty members in science and engineering, to introduce them to ethical concerns and to prepare them to mentor their own students on research integrity.
Intellectual merit: The project will engage graduate students and postdoctoral research associates in the analysis of the serious and complex ethical issues that face all scientists and engineers in the course of their careers.
Broader impacts: The project will produce (1) graduate students who know how to address ethical issues; (2) postdoctoral research associates who not only understand these issues, but know how to teach on these issues and can expand the impact of the project across Maryland, the United States, and beyond; (3) new faculty members who appreciate the importance of research integrity and can apply it in the training of their own students; and (4) extension of the courses developed at College Park to other campuses in Maryland.

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Popper
This award to University of Maryland provides support for scientific participant travel and expenses for participation by US scientists in a conference to be held in Denmark in 2007 addressing issues related to the impacts of sound on marine fauna. The topic of anthropogenic sound in the marine environment has become an issue of significant regulatory and public concern, both domestically and internationally, but the scientific information available to make informed decisions regarding actual impacts is quite limited. In conjunction with separate support from other US and international groups, support is provided here to bring experts together to address the scientific findings and additional needs on this topic, and to produce a set of publications that help define the state of knowledge on the use of sound by various marine organisms, processes and changes in underwater acoustics and ambient noise, sources of anthropogenic noise and their characteristics, effects of noise on various animal groups, and the current national and international regulatory framework for use of noise in the marine environment.
Broader Impacts: Support provided here will improve efforts to assess potential impacts of natural and anthropogenic sounds on marine animals. Many human activities, including scientific research, introduce sound into the marine environment intentionally or incidentally, and a clear understanding of the nature of such impacts on organisms is an important element in our effort to properly assess potential environmental impacts based on the best available science.
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The PI requests funding to partially support two activities related to the effects of noise. The first is The Second International Conference on the Effects of Noise on Aquatic Life workshop which will take place in the city of Cork, in the Republic of Ireland August 15-20, 2010. The second is a Working Group on the Effects of Noise on Fish and Turtles. The Working is in the final stages of completing a draft manuscript that lays out the data in the literature and research recommendations.

Broader Impacts

The goal of the conference is to draw together new knowledge on the importance of underwater sound to animals and to the effects of sounds upon them, whether those sounds occur naturally, are made by the animals themselves, or result from human activities. The product of the Working Group will provide the first consensus document, produced by the scientists who have been doing much of the work on fish and turtle noise effects and fish and turtle bioacoustics, on the value of the data collected to date, and on their usefulness in setting criteria.

The PI requests funding for The Third International Conference on the Effects of Noise on Aquatic Life will take place in Budapest, Hungary, August 11-16, 2013. This meeting follows two successful meetings in Nyborg (2007) and Cork (2010), both of which NSF provided partial support. The major goal of the conference will be to define the current state of knowledge on the impact of underwater noise and, in particular, explore the progress made in this field in the three years since the second Conference. The third Conference will place strong emphasis on recent research results, the sharing of ideas, discussion of experimental approaches, and analysis of regulatory issues.

Broader Impacts

The meeting should help shape future research and understanding of effects of noise on aquatic life. Based on experience from the earlier meetings, the interactions and networking that took place has resulted in new collaborations between investigators, a broader understanding of the science that can be used in their work by regulators, a better understanding of regulatory issues by scientists and industry, and substantial cross-fertilization of experimental design and approaches by investigators.

This award provides partial support for The Fourth International Conference on the Effects of Noise on Aquatic Life will take place in Dublin, Ireland, July 10-16, 2016. This meeting follows the very successful meetings on the same general topic held in Nyborg, Denmark (2007), Cork,Ireland (2010), and Budapest, Hungary (2013). The major goal of the conference will be to define the current state of knowledge on the impact of underwater noise and, in particular, explore the progress made in this field in the three years since the previous Conference. The extension of renewable energy and petroleum projects to offshore waters, the prospect of industrial development of previously pristine waters (e.g., polar regions), and recent proposals to open new areas for petroleum lease (e.g., US Atlantic coast) has resulted in greater interest in the effects of underwater noise on aquatic life on the part of regulators, industry, and scientists.

The meeting should help shape future research and understanding of effects of noise on aquatic life. Based on experience from the earlier meetings, the interactions and networking that took place has resulted in new collaborations between investigators, a broader understanding of the science that can be used in their work by regulators, a better understanding of regulatory issues by scientists and industry, and substantial cross-fertilization of experimental design and approaches by investigators.