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At least 145 records · Page 8

Data for Mitochondrial ATP Generation is More Proteome Efficient than Glycolysis

Metabolic efficiency profoundly influences organismal fitness. Heterotrophs, from yeast to mammals, derive usable energy primarily through glycolysis and respiration. While respiration is more energy-efficient, some cells favor glycolysis even when oxygen is available (aerobic glycolysis, Warburg effect). A leading explanation is that glycolysis is more efficient in terms of ATP production per unit mass of protein (i.e. faster). Through quantitative flux analysis and proteomics, we find however that mitochondrial respiration is actually more proteome-efficient than aerobic glycolysis. This is shown across yeasts, T cells, cancer cells, and tissues and tumors in vivo. Instead of aerobic glycolysis being valuable for fast ATP production, it correlates with high glycolytic protein expression, which is valuable for hypoxic growth. Aerobic glycolytic yeasts do not excel at aerobic growth, but outgrow respiratory cells in oxygen limitation. Thus, aerobic glycolysis emerges from cells maintaining a proteome conducive to both aerobic and hypoxic growth.

Metabolomics↗

Host population dynamics influence Leptospira spp. transmission patterns among Rattus norvegicus in Boston, Massachusetts, US

Leptospirosis (caused by pathogenic bacteria in the genus Leptospira ) is prevalent worldwide but more common in tropical and subtropical regions. Transmission can occur following direct exposure to infected urine from reservoir hosts, or a urine-contaminated environment, which then can serve as an infection source for additional rats and other mammals, including humans. The brown rat, Rattus norvegicus , is an important reservoir of Leptospira spp. in urban settings. We investigated the presence of Leptospira spp. among brown rats in Boston, Massachusetts and hypothesized that rat population dynamics in this urban setting influence the transportation, persistence, and diversity of Leptospira spp. We analyzed DNA from 328 rat kidney samples collected from 17 sites in Boston over a seven-year period (2016–2022); 59 rats representing 12 of 17 sites were positive for Leptospira spp. We used 21 neutral microsatellite loci to genotype 311 rats and utilized the resulting data to investigate genetic connectivity among sampling sites. We generated whole genome sequences for 28 Leptospira spp. isolates obtained from frozen and fresh tissue from some of the 59 positive rat kidneys. When isolates were not obtained, we attempted genomic DNA capture and enrichment, which yielded 14 additional Leptospira spp. genomes from rats. We also generated an enriched Leptospira spp. genome from a 2018 human case in Boston. We found evidence of high genetic structure among rat populations that is likely influenced by major roads and/or other dispersal barriers, resulting in distinct rat population groups within the city; at certain sites these groups persisted for multiple years. We identified multiple distinct phylogenetic clades of L. interrogans among rats that were tightly linked to distinct rat populations. This pattern suggests L. interrogans persists in local rat populations and its transportation is influenced by rat population dynamics. Finally, our genomic analyses of the Leptospira spp. detected in the 2018 human leptospirosis case in Boston suggests a link to rats as the source. These findings will be useful for guiding rat control and human leptospirosis mitigation efforts in this and other similar urban settings.

Stone, Nathan E.↗

A robust method of nuclei isolation for single-cell RNA sequencing of solid tissues from the plant genus Populus

Single-cell transcriptome analysis has been extensively applied in humans and animal models to uncover gene expression heterogeneity between the different cell types of a tissue or an organ. It demonstrated its capability to discover key regulatory elements that determine cell fate during developmental programs. Single-cell analysis requires the isolation and labeling of the messenger RNA (mRNA) derived from each cell. These challenges were primarily addressed in mammals by developing microfluidic-based approaches. For plant species whose cells contain cell walls, these approaches have generally required the generation of isolated protoplasts. Many plant tissues’ secondary cell wall hinders enzymatic digestion required for individual protoplast isolation, resulting in an unequal representation of cell types in a protoplast population. This limitation is especially critical for cell types located in the inner layers of a tissue or the inner tissues of an organ. Consequently, single-cell RNA sequencing (scRNA-seq) studies using microfluidic approaches in plants have mainly been restricted to Arabidopsis roots, for which well-established procedures of protoplast isolation are available. Here we present a simple alternative approach to generating high-quality protoplasts from plant tissue by characterizing the mRNA extracted from individual nuclei instead of whole cells. We developed the protocol using two different plant materials with varying cellular complexity levels and cell wall structure, Populus shoot apices, and more lignified stems. Using the 10× Genomics Chromium technology, we show that this procedure results in intact mRNA isolation and limited leakage, with a broad representation of individual cell transcriptomes.

54 ENVIRONMENTAL SCIENCES↗

Assessing springtime vertebrate prey of sympatric mesopredators in the southeastern United States using metabarcoding analysis

Coyotes (Canis latrans) colonized the eastern United States over the last century and formed a 3-species predator guild with bobcats (Lynx rufus) and gray foxes (Urocyon cinereoargenteus) across much of the southeastern United States. Diets among the three species vary along with respective impacts on game species such as white-tailed deer (Odocoileus virginianus) and wild turkeys (Meleagris gallopavo). To determine predation impacts on vertebrate prey and dietary overlap in consumption of prey items, we assessed diets of coyote, bobcat, and gray fox during spring, coinciding with white-tailed deer fawning and wild turkey nesting and brood rearing. We sampled across three sites along the Savannah River in South Carolina from mid-May through mid-June of 2020–2021. We collected 180 scat samples along 295.9 kilometers (71.1–122.4 km/site) of unpaved secondary roads and used DNA metabarcoding to determine vertebrate diet items. We identified predator species of scat using DNA metabarcoding and species-specific mtDNA fragment analysis (153 were coyote, 20 bobcat, and seven gray fox). Overall, we found evidence that two species, coyote and bobcat, consumed deer while all three consumed turkeys. Frequency of deer in the diet varied across sites for coyotes from 62–86% and wild turkey was present with a frequency of occurrence of 9% for coyotes, 5% for bobcats, and 14% for gray fox. Vertebrate diet specialization was evident across predator species with high frequency of deer in coyote diets, rabbits and small mammals in bobcat diets, and herpetofauna in gray fox diets. During deer fawning and wild turkey nesting and brood rearing, dietary overlap appears to be mediated by disparate selection of prey items, which reduced competition among coyotes, bobcats, and gray foxes. Use of DNA metabarcoding may augment our understanding of dietary preferences within this predator guild by providing increased resolution of diet composition among important game species.

59 BASIC BIOLOGICAL SCIENCES↗

Observations of marine animal interactions with a small tidal turbine

The risk of collisions between animals and operating tidal turbines remains a concern in the scientific and regulatory communities. A sensor package including optical cameras was deployed to monitor animal interactions with a small-scale (1 m 2 ) cross-flow tidal turbine. The turbine was deployed in Washington State, USA for 141 days at a site with peak flow speeds of 2.5 m/s. We analyze optical camera imagery spanning 109 days of turbine operation. The analyzed images contain 1044 observations of fish, fish schools, seabirds, or seals in the vicinity of the turbine. No instances of collision with seabirds or seals were observed. Seabirds were only observed during daylight hours and while the turbine was stationary. Both seals and fish were observed during both day and night and while the turbine was stationary and rotating. Four fish were observed colliding with the moving turbine and in all but one case the animals swam away following the collision. Over the same period of time, over fifty times more fish (224 individual fish and 5 fish schools) were observed passing the moving turbine without collision. Fish encounters were likely under counted due to the difficulty in discerning small fish from plant matter in the water column. These observations represent the first optical camera imagery showing fish, bird, and marine mammal interactions with a tidal turbine in North America. In addition to quantitative and qualitative discussion of the implications of our observations for collision risk, we discuss lessons learned on sampling schemes and deployment of machine learning for detection of animals to inform future data collection strategies in future monitoring campaigns.

16 TIDAL AND WAVE POWER↗

The Monkeys and Parrots of Gold Rush-era California

As immigrant gold miners migrated en masse to San Francisco and northern California during the Gold Rush-era (ca. 1849–1855), they experienced new animals. Stopping in ports throughout Central and South America, these argonauts saw, felt, smelled, heard, and occasionally consumed, mammals, birds, reptiles, and many more creatures, which were wholly exotic to those species found at home. Two types of animals that the Gold Rush populace encountered during this era include parrots and monkeys. Although found throughout tropical environments in areas far distant from northern California, these animals became quickly imported to San Francisco during the early 1850s. A wild, turbulent Gold Rush-era helped facilitate the importation of these exotic animal types, both for comfort and entertainment, as they helped provide a source of companionship for miners unaccustomed to the shock of 1850s northern California.

59 BASIC BIOLOGICAL SCIENCES↗

AmeriFlux US-BFS Robert J. Bernard Biological Field Station

This is the AmeriFlux version of the carbon flux data for the site US-BFS Robert J. Bernard Biological Field Station. Site Description - The Bernard Field Station (BFS), an 86-acre academic resource of the Claremont University Consortium, is situated within the campus of The Claremont Colleges in Claremont, CA. Its principal habitats comprise coastal sage scrub, Riversidian alluvial fan scrub, live oak forest, and grassland. The eddy covariance tower is located in the 'neck' of the field station, home to a protected area of Coastal Sage Scrub, dominated by Artemisia californica. BFS is within walking distance of Harvey Mudd College, serving as an invaluable outdoor laboratory resource for the students. With extensive flora and fauna, including over 170 species of birds, 27 species of mammals, and more than 700 species of insects, it embodies a significant ecological haven in the region that is otherwise Southern California suburbs.

Kavassalis, Sarah [Harvey Mudd College]↗

Opiate Sensitivity in Fruit Flies

Substance use disorder is a debilitating clinical condition in which behavioral dependence results from biological, environmental, genetic, and psychosocial factors. An epidemic surrounding the use and abuse of opioids is ravaging the world. While considerable efforts have explored the social drivers of addiction, a deeper understanding of biological causes and genetic vulnerabilities, preventative interventions, and effective treatments, have all proven elusive. This perspective article aims to remind readers that addictive natural compounds such as cocaine, nicotine, cathinone, or morphine, evolved as defensive metabolites to deter insect herbivory. The molecular mechanisms underlying motivational seeking and learning/reward show remarkable conservation since their early emergence in bilateral metazoans. An extended coevolutionary arms race subsequently weaponized these compounds into disruptors of learning, motivation, and incentivized attention. When plant chemical defenses attack insect physiology, humans are rendered susceptible due to strong conservation in the underlying molecular machinery. This perspective addresses the paradox that opiates were shaped to target insect neuropharmacology, even though this taxon appears to lack the recognized opioid receptor clade of mammals. We argue that the link is to be found in the allatostatin receptor, a basal ortholog of opioid receptors. Moreover, preliminary evidence indicates that morphine reduces Drosophila feeding and locomotion, concordant with a purported role as a defensive compound reducing herbivory. An implementation via allatostatin-mediated mechanisms is likely. This research argues for a broader heuristic perspective of substance abuse and a recognition of the evolutionary constraints that have likely shaped the biological drivers of opioid sensitivity and of its behavioral targets.

59 BASIC BIOLOGICAL SCIENCES↗

Exposure to ionizing radiation disrupts normal epigenetic aging in Japanese medaka

Alterations to the epigenome are a hallmark of biological aging and age-dependent patterning of the DNA methylome (“epigenetic aging”) can be modeled to produce epigenetic age predictors. Rates of epigenetic aging vary amongst individuals and correlate to the onset of age-related disease and all-cause mortality. Yet, the origins of epigenetic-to-chronological age discordance are not empirically resolved. Here, we investigate the relationship between aging, DNA methylation, and environmental exposures in Japanese medaka (Oryzias latipes). We find age-associated DNA methylation patterning enriched in genomic regions of low CpG density and that, similar to mammals, most age-related changes occur during early life. We construct an epigenetic clock capable of predicting chronological age with a mean error of 61.1 days (~8.4% of average lifespan). To test the role of environmental factors in driving epigenetic age variation, we exposed medaka to chronic, environmentally relevant doses of ionizing radiation. Because most organisms share an evolutionary history with ionizing radiation, we hypothesized that exposure would reveal fundamental insights into environment-by-epigenetic aging interactions. Radiation exposure disrupted epigenetic aging by accelerating and decelerating normal age-associated patterning and was most pronounced in cytosines that were moderately associated with age. These findings empirically demonstrate the role of DNA methylation in integrating environmental factors into aging trajectories.

60 APPLIED LIFE SCIENCES↗

Coronaviruses: Epidemics and Pandemics

Coronaviruses constitute a large group of viruses that circulate in domestic and wild mammals and birds (human, cat, dog, pig, mouse, rat, bat, cattle, deer, giraffe, llama, sheep, elk, caribou, and birds). Seven of them have made their way into humans. This presentation gives an overview of three coronavirus epidemics with an emphasis on Severe Acute Respiratory Syndrome Coronavirus-2 (SARS CoV-2) which causes coronavirus disease -2019 (COVID-19).

59 BASIC BIOLOGICAL SCIENCES↗

2020 State of the Science Report, Chapter 3: Collision Risk for Animals around Turbines

The potential for marine animals to encounter and collide with turbines, especially tidal and river turbines, along with the biological, ecological, and regulatory consequences of any such interactions, remain active areas of research and topics of global interest. Uncertainty and knowledge gaps associated with collision risk continue to present challenges within consenting/permitting (hereafter consenting) processes for turbine developments. Consequently, collision risk continues to be the focus of significant research effort, which in recent years has included environmental monitoring of operational devices and arrays. This chapter addresses the overall progress and growth in knowledge across this topic area, and specific progress related to marine mammals, fish, and seabirds. https://tethys.pnnl.gov/publications/state-of-the-science-2020-chapter-3-collision

13 HYDRO ENERGY↗

Tribal Revegetation Project Final Project Report: 92-Acre Area, Area 5 Radioactive Waste Management Complex, Nevada National Security Site, Nevada

Nuwu (Southern Paiute), Newe (Western Shoshone), and Nuumu (Owens Valley Paiute) are linguistically related, Numic-speaking peoples who are part of the broader Uto-Aztecan language group. Numic peoples view the land as a holistic, living, sentient being with feelings and purpose. The land is personified with human characteristics and it needs to be experienced to be understood through “learning by doing.” Numic peoples do not support ground disturbing activities within their ancestral lands, including activities tied to the storage of low-level radioactive waste or classified materials on the NNSS, which they view as culturally inappropriate. These deep-rooted ancestral connections are the impetus for reinforcing Numic responsibility for healing disturbed areas by integrating respect and patience with consistent Tribal interaction. Tribal Ecological Knowledge (TEK) is the science of describing Tribal approaches for understanding natural resources. Numic TEK is embedded in traditional teachings learned incrementally over time though experience and it evolves through lessons learned and responses to environmental changes over millennia. Therefore, TEK can broaden and enhance Western scientific knowledge associated with revegetation, especially in highly disturbed areas. The project blended TEK with Western scientific ecological methods to create a vegetative cover within test plots on the 92-Acre Area located at the Radioactive Waste Management Complex (RWMC) located in Area 5 on the Nevada National Security Site (NNSS). The vegetated test plots were systematically created for the Department of Energy (DOE) in tandem with the existing Federal Facilities Agreement and Consent Order (FFACO) with the Nevada Department of Environmental Protection (NDEP). Three previous contractor-lead attempts at revegetation, one targeting full cover revegetation and two targeting test plot revegetation, did not achieve the anticipated results at this location. When presented to the 16 American Indian Tribal nations and affiliated groups with cultural and historical ties to the NNSS, the group appointed a Tribal Revegetation Committee (TRC) that included six expert Tribal knowledge holders to collaborate with an ethnoecologist/cultural anthropologist and two biologists. Project design, planning, seed and outplant selection, spiritual land preparation, and methodology were guided by the TRC and an ethnoecologist/cultural anthropologist from Portland State University (PSU) and biologists from Desert Research Institute (DRI). Using TEK, the TRC recommended a specific seed mixture that contained nine native plant species and three species of outplants. The revegetation effort included preparing and planting thirty 10 m × 10 m (32.8 ft × 32.8 ft) seeded plots, twelve of which also included outplants; and eight 10 m × 100 m (32.8 ft × 328 ft) plots that only received outplants, all atop a waste cell cover. The TRC and the project team creatively adapted TEK with Western scientific methods so that the planned revegetation efforts could occur within the safety and security parameters of the RWMC. Test plots were subjected to one of five soil treatments with varying combinations of straw or mulch applications, soil amendment, and/or outplant planting and one of two watering treatments (watered or unwatered). Planting was divided into two events: one in the fall season and another during the subsequent spring season based on TEK and a corresponding recommendation from the TRC. Monitoring and spiritual management was conducted by the TRC to evaluate plant progress on a monthly basis (in conjunction with the biologist and anthropologist) during each respective growing season for a period of three years after planting. This approach allowed Tribal members the opportunity to conduct traditional blessings and other culturally appropriate activities to restore balance to the land in accordance with Tribal protocols. Following TEK-guided methods, successful plant establishment from seed stock and outplants was observed in many plots. Overall, plots planted in the spring, as recommended by the TRC, showed higher rates of outplant survival and native seedling emergence than those planted in the fall. This finding is significant because it is contrary to the original guidance and previous approaches provided for planting in this region. The TRC believes the frequent co-occurrence of native seedlings near surviving outplants indicates an important symbiotic relationship understood by Tribal communities and overlooked by others. Watered outplants displayed much higher survivability than unwatered plants, even after watering was reduced after the plants were established. Soil amendments and mulch created higher densities of native plants from seed. Many native seedlings showed significant delays in germination, which is considered a normal adaptation to desert climates. Some native plants did not germinate until the third growing year, whereas others germinated during the first growing year, which demonstrates the complexity of the desert environment. Evidence of native insects, reptiles, mammals, and birds, as well as native plants that were not part of the planted species, were noted and considered culturally significant. Despite the presence of non-native plants, native outplants continued to thrive and the incidence of native plant germination from seed increased over time. These successful revegetation results where previous efforts were unsuccessful reinforce the importance of integrating regionally appropriate, TEK-guided methodology with Western science to achieve positive results and the necessity of integrating Tribal involvement in all stages of the revegetation effort. Expanded approaches coupled with Tribal knowledge and tools from Western science addressed a complex problem tied to revegetating atop a low-level radioactive waste cell. The level of Tribal participation serves as a progressive model for building collaborative relationships and addressing ecological challenges on the NNSS.

54 ENVIRONMENTAL SCIENCES↗

Environmental Effects Assessment for Proposed Offshore Wind Farm off the Coast of Grays Harbor, Washington

Grays Harbor Wind LLC (GHW) is proposing to develop a floating offshore wind farm offshore of west Grays Harbor County, Washington (Grays Harbor). The proposed GHW Offshore Wind Project (Project) would entail construction, installation and operation of a 1,000-megawatt (MW) offshore wind farm consisting of approximately 75 floating units, each containing a floating foundation and wind turbine generator (WTG). The Project location is approximately 25 miles (21.7 nautical miles [nmi]) offshore west of Grays Harbor, at waters depths of 360 to 700 feet. The Pacific Northwest National Laboratory (PNNL) was contracted by Herrera Environmental Consultants, Inc. on behalf of GHW to carry out this preliminary scoping study to evaluate baseline conditions and potential effects on fish and marine mammals from development and operation of a floating offshore wind farm installed within a designated area off the coast of Washington1. Floating offshore wind units installed in an ocean environment as part of the Project would interact with marine wildlife. This Study report provides an initial data aggregation and analysis, using publicly available data, of the Project effects, both negative and positive, on the marine environment. The scope of this assessment is limited by the fact that the Project development is presently at the conceptual level. Data on marine organisms were aggregated and evaluated; however seabirds were evaluated by Herrera Environmental Consulting and, along with bats, were not included in the scope of this study. Significant additional work is necessary to characterize ocean, seafloor, and environmental conditions; select appropriate floating offshore wind technologies; identify construction methods and locations; and assess facility locations, including electrical interconnection. Data aggregation and analysis of seabirds and bats is also needed. Evaluation of the full range of potential environmental effects would be conducted following an award of a lease from the Bureau of Ocean Energy Management (BOEM) as part of the leasing, National Environmental Policy Act (NEPA)/State Environmental Policy Act (SEPA) environmental review and permitting processes. While this initial data aggregation and analysis uses best available public scientific information and current assumptions about the Project configuration, the effects discussed herein are based on the status of review to-date and may change as Project-specific details are developed.

17 WIND ENERGY↗

Brain Connectivity Workshop Series Report

The National Institutes of Health (NIH) BRAIN Initiative aims to accelerate development and application of technologies that show how individual brain cells in complex neural circuits interact at the speed of thought and action. Complete wiring diagrams of the mammalian brain will revolutionize the capabilities of researchers to formulate and test models of how activity within brain circuits drives coordinated function and behavior. A top priority for the BRAIN Initiative, motivated by the strategic guidance of the BRAIN 2.0 Working Group report, is to muster the resources and foster the collaborations needed to generate these wiring diagrams at the level of long-range projections (“projectomes”) and synapses (“connectomes”) in whole mammalian brains, including those from rodents, humans, and other large-brained mammals. Achieving the scale and resolution necessary to characterize and use these projectomes and connectomes will not only push the boundaries of current imaging and computational capabilities to generate faster, cheaper, and more scalable technologies, but also drive broad innovation in data science, artificial intelligence, and machine learning.

42 ENGINEERING↗

A Miniaturized Long-Life, Low-Frequency Acoustic Transmitter for Fish Tracking in Marine Environment - CRADA 447

Advance state-of-art technologies for tracking marine mammals, fish, sea turtles, and other protected species, based on acoustic telemetry. Objectives include: (1) develop a new acoustic transmitter that has a greatly improved detection range and service life while being significantly smaller and lighter than the existing commercial counterparts; and (2) reach a TRL of 5 so the technology is ready for organizations interested in small- or medium-scale field trials.

42 ENGINEERING↗

TEAMER: DAISY Flow-noise Testing (Abstract)

Evaluating impacts of marine energy devices is generally difficult given the dynamic environment where these devices need to be placed. The University of Washington’s (UW) DAISY (The Drifting Acoustic Instrumentation SYstem) is designed to measure radiated noise around marine energy converters operating in energetic waves and currents. In currents, a primary limitation for measurement fidelity at low frequencies (< 100 Hz) is the potential for non-propagating “flow-noise” to mask propagating sound and inflate estimates of the radiated noise from marine energy converters at frequencies that overlap with hearing sensitivities of fish and some marine mammals. While free-drifting measurements help to minimize the relative velocity that produces flow-noise, significant levels were still observed during initial DAISY tests. This motivated the development of a fabric “flow shield” around the hydrophone that disrupts both flow-noise generation mechanisms proposed by the initial tests. First, the flow shield is a source of substantial drag which keeps the hydrophone package moving with the approximate velocity of the surrounding water, compensating for differential wind or current forcing on the surface expression. By minimizing relative velocity around the hydrophone, turbulence shed by the hydrophone is also minimized. Second, the flow shield creates a largely quiescent pocket around the hydrophone, minimizing advection of free stream turbulence over the hydrophone element. Field data collected in these experiments will test effectiveness of these flow shields and provide quantitative data use and deployment.

16 TIDAL AND WAVE POWER↗

'Omics and Big Data in Harmful Algal Bloom Research

Phytoplankton, a group including eukaryotic microalgae and cyanobacteria, play a crucial climate role converting CO 2 into organic carbon through global primary production. They support a wide range of life, both freshwater and marine, from zooplankton to fish and mammals. While they are essential in nutrient cycles, certain phytoplankton species can proliferate excessively under favorable conditions, leading to harmful algal blooms (HABs) that pose significant threats to human and ecosystem health through the toxins they produce.

59 BASIC BIOLOGICAL SCIENCES↗

CRCNS US-France Research Proposal: Collaborative Research: Encoding reward expectation in Drosophilia

The fruit fly Drosophila melanogaster has been a valuable model for investigating the genetic and neural bases that underlie learning and memory. Early and most current studies use basic behavior conditioning protocols to study learning in controlled laboratory settings. More recently, the ability to transgenically manipulate many of the brain neurons in the fruit fly with exquisite specificity, and the recent knowledge of the synaptic ‘connectome’ of the fruit fly brain, makes these animals almost unique as a comprehensive model for studies of learning, memory and motivated behavior. In fact, the connectome has revealed many types of new connections that had until now been overlooked. Within this context, the thesis of this proposal is that studies of learning and memory will be greatly enhanced by using more sophisticated means for evaluating memory representations, such as have been developed in vertebrates, and combining those studies with information from the connectome guided by computational modelling. We propose to push beyond the boundaries of existing conditioning protocols for fruit flies to investigate more complex memory representations. In particular, we will investigate the function of reinforcement pathways in relation to the absence of expected reinforcement. More specifically, we propose a series of experiments designed to investigate the memory representations in fruit flies when an expected consequence of a Conditioned Stimulus (CS) fails to occur. Although studies have evaluated how this failure can establish extinction memory for the CS, our studies will go beyond studying extinction. Specifically, we predict that in Drosophila when a CS is associated with a failed expectation of an appetitive food reinforcement it will acquire aversive value, and vice versa for a failed expectation of an aversive reinforcer. We combine these studies with manipulations of reinforcement pathways in the CNS inspired from the connectome, iteratively knitted in with established computational models. Intellectual Merit: The concept of reinforcement expectation and incentive contrast have been influential in the development of studies of associative learning in mammals. These questions are particularly challenging to answer in vertebrates because they require exquisite cellular, temporal, and genetic specificity of experimental manipulations. The recent development of work with identified neurons and their connectomes makes the larval and adult fly brains ripe as models for pushing our understanding of neural bases for these higher- order conditioning phenomena. Broader Impacts: Public health: These analyses and the conceptual framework of prediction error processing underlying them have a profound impact on our understanding of reinforcement-related behavior in humans, including monetary rewards and the mnemonic consequences of traumatic experiences, and for pathologies of the dopamine reinforcement system. Educational: This project will provide interdisciplinary training for postdoctoral researchers, Ph.D. and undergraduate students. The PIs will act as co-supervisors or mentors of students working in the different labs via face-to-face and internet-based technologies. We will also work with ASU’s award-winning Ask- A-Biologist program. This is an online science program designed to enrich the learning experiences of students of all ages and to provide classroom material for use by K-12 teachers. We will develop an extension of a game developed under a prior NSF award, and the new game will include modules to teach K-12 students about how insects learn. We will also integrate into the AAB site a program developed by a collaborator (B Gerber) at the Leibniz Institut für Neurobiologie, Magdeburg, and now in use in schools in Germany, to teach K-12 students how to train animals using the fruit fly larval learning paradigm. Underrepresented groups: All PIs will work with their university offices of Academic Diversity and Equal Opportunity for reaching underrepresented students.

59 BASIC BIOLOGICAL SCIENCES↗