Richard W. Tsien, D. Phil. - US grants

Channels carrying slow inward Ca current are vital to the rhythm and contraction of the healthy heart, and play an important part in arrhythmias. We will study the basic properties of cardiac Ca channels, their inhibition by "Ca antagonists", and their modulation by beta-adrenergic and cholinergic neurotransmitters. We will take advantage of two new and powerful approaches: the dialyzed cell method and the patch clamp technique. As we have shown in recent papers, it is possible to use these methods to record Ca channel activity from single heart cells, or even individual channels. The methods will also allow delivery of drugs, enzymes, or putative messenger molecules to the inside surface of the cell membrane. These approaches will help answer some fundamental questions. What is the ion flux through a single open Ca channel? How many functional Ca channels does a single cell have, and does the number change under the influence of neurohormones? Does the Ca channel open by a single one-step process or a multi-step mechanism? How is Ca channel inactivation affected by intracellular Ca and membrane potential? Resolving these unsettled physiological issues will help answer some important pharmacological questions. Although "Ca antagonists" have already achieved clinical prominence, their basic mechanism(s) of action remain poorly understood. Do such drugs block current flowing outward through the channel as well as inward (as lidocaine or tetrodotoxin do in the case of the Na channel)? Do they act from inside the cell? Do they really compete with Ca ions, as the term "antagonist" implies? Do they block open or inactivated channels preferentially? In answering these questions, we might find important differences between verapamil, diltiazem and nifedipine, and the classes of drug they represent, while also gaining basic insights into Ca channel function. We will pursue preliminary hints that isoproterenol increases Ca entry through changes in the percentage of time individual Ca channels stay open, and perhaps also through a slowing of Ca channel inactivation. We will find out whether dimetrically opposite mechanisms are at work when acetylcholine decreases Ca current. The possible involvement of cyclic nucleotides and protein phosphorylation in mediating the actions of beta-adrenergic and cholinergic agents will be tested at the level of single Ca channels.

Calcium entry via Ca channels contributes to a wide range of neuronal functions including pacemaker activity, spike electrogenesis, transmitter release and neurite outgrowth. Multiple components of Ca current have been found in neurons as well as other excitable cells. For example, our previous patch clamp recordings demonstrated that chick dorsal root ganglion (DRG) cells contain three types of Ca channels (T, N and L-type) which differ in single channel conductance, gating kinetics, and pharmacological sensitivity. This proposal describes studies of the properties, distribution and functional roles of the various types of Ca channel in cultured neurons, hippocampal slices and isolated synaptic endings. (1) We will continue development of selective agents for dissecting channel function, having seen selective effects of Bay K 8644 and wCgTX, a 27-amino acid marine snail toxin. We will analyze effects of organic drugs (verapamil, diltiazem, nifedipine, etc), peptide toxins (wCgTx, atrotoxin, B-leptinotarsin), and agents that might selectively block T-type channels. (2) We will explore differences amongst the channel types in mechanisms of ion permeation and gating. Modulation of channel activity by neurotransmitters or internal messengers will be analysed at the level of single Ca channels. (3) The distribution of various Ca channel types on cell bodies, dendrites, and growth cones of cultured neurons will be studied with on-cell, outside-out and loose patch recordings. The electrophysiological findings will be compared with localization of N- and L-type channels by autoradiography with 125I-wCgTx. (4) Participation of Ca channel types in electrical activity of the hippocampus will be studied with patch and chopped clamp recordings from hippocampal pyramidal and granule cells. We will characterize the dendritic Ca channels that underly fast pre-potentials and epileptiform activity, and look for modulatory effects of C-kinase on Ca channels that might accompany long-term potentiation. (5) The involvement of L-type orrN-type Ca channels in mediating Ca entry and transmitter release will be studied in presynaptic terminals of neuromuscular junctions, cultured neurons, brain slices, and synaptosomes. Activity of synaptosome Ca channels will also be recorded following incorporation in planar bilayers.

biophysics; membrane permeability; ion transport; biochemistry;

DESCRIPTION (provided by applicant): Many long-standing questions about fundamental mechanisms of neurotransmission recur in current attempts to understand how channelopathies give rise to synaptic defects. A striking overlap between cell biological and pathophysiological questions arises in the case of P/Q-type voltage-gated Ca2+ channels, whose pore-forming alpha1A subunit (also called alphaCa-v2.1) plays a dominant role in supporting voltage-gated presynaptic Ca2+ entry and fast excitatory and inhibitory neurotransmission. Working together with N- and R-type Ca2+ channels, P/Q-type channels contribute the majority of presynaptic Ca2+ entry and also emerge as the principal molecular target in known calcium channelopathies, including monogenetic forms of such neurological disorders as ataxia, epilepsy and headache. This project will investigate fundamental issues about P/Q-type channels and neurotransmission that may also illuminate pathophysiological mechanisms. Working with dissociated neuronal cultures from alpha1A -/- mice, we will examine the ability of various mutant human CCIA subunits to support Ca2+ channel activity and synaptic transmission. We will test the hypothesis that the relative efficacy of P/Q-type channels is governed by P/Q-selective "slots" that cap the contribution of P/Q-type channels even when they are greatly overexpressed. Specific questions to be addressed include the following: Do type selective slots for presynaptic N- and R-type Ca2+ channels co-exist along with those functionally defined for P/Q channels? If type-specific slots exist, does deficiency in Ca2+ channel function translate into an overall reduction in nerve terminal Ca2+ influx? What is the basic topography of Ca2+ channels in small presynaptic terminals? Where does the competition for slots occur and what is its molecular basis? What are the cell-biological or biophysical mechanisms by which human disease mutations affect Ca2+ channel function and higher-order pathophysiology? Why do P/Q channel diseases show a dominant negative inheritance? What are the homeostatic mechanisms that compensate for loss of P/Q channel function and what is their role in governing the overall outcome of the disease?

No parent abstract provided.

Our experiments are motivated by the conviction that progress in research on LTP and other forms of synaptic plasticity must proceed hand with gaining a better understanding of fundamental characteristics of synaptic transmission. Basic knowledge about the physiological factors determining the reliability of transmission and the amplitude of the postsynaptic response in CNS neurons is still lacking. Non-uniform behavior from one synapse to the next poses great problems for analysis of synaptic transmission in preparations where the number and location of stimulated synapses is not known. To overcome these problems, we will use two new approaches to study the behavior of single visualized synapses in hippocampal cultures: first, the use of differential uptake of antibodies against a lumenal protein marker, synaptotagmin, to monitor changes in presynaptic activity independently of postsynaptic responsiveness (Malgaroli et al., Science, in press), and second, the focal stimulation of transmitter release from single presynaptic boutons or glutamatergic responses from individual postsynaptic receptor clusters (Lie et al., nature, in press). These strategies will help us to settle some long- standing questions about normal synaptic transmission, and to study the enduring modification of transmission during LTP. Our specific goals are as follows: (1) Mechanisms governing the reliability of the synaptic response (failures or successes) and variability in quantal response size will be studied at the level of single, visualized synapses, (2) LTP will be induced at single synapses and the spread of potentiation and the relative importance of presynaptic and postsynaptic changes will be assessed by direct methods. (3) The locus and mechanisms of late long-term potentiation (L-LTP) -- potentiation lasting many hours -- will be studied with cell biological and morphological approaches, (4) Signaling mechanisms for initiation and maintenance of various forms of LTP will be analyzed separately with regard to their pre- and postsynaptic components.

DESCRIPTION: Neuronal Ca2+ entry via voltage-gated Ca2+ channels and NMDA receptor Channels provides a critical link between synaptic activity and changes in gene expression. Despite the importance of communication from synapse to nucleus, relatively little is known about basic mechanisms. A key target of such signaling is the nuclear transcription factor CREB undergoes phosphorylation at Ser133, it can turn on transcription and thereby activate a host of CRE-dependent genes. Recently, our group has uncovered a novel signal transduction pathway that links CA2+ entry to rapid (<1 min) CREB phosphorylation. The signaling is initiated by opening of L-type (but not P/Q or N-type) CA2+ channels, and depends critically on the translocation of calmodulin from sites near the surface membrane to within the nucleus itself, where it activates nuclear CaMKIV, a potent CREB kinase. In this project, the investigators propose to study cellular and molecular aspects of the synapse t nucleus signaling. What gives L-type channels the specific ability to initiate CaM translocation? What are the spatiotemporal properties of CaM signaling following general or focal stimulation? What molecular mechanisms support translocation of CaM to the nucleus and stabilization of its Ca2+-bound form? How do the multiple targets of CaM (e.g. kinase kinase, kinase, phosphatase) interact together to control CREB phosphorylation levels and downstream gene expression? What are the consequences of disrupting fast CaM signaling in hippocampal slices? The investigators will approach these questions as an extension of our long-term interest in the functional impact of diverse Ca2+ channels, using a potent combination of techniques, including patch clamp and calmodulin distributions.

[unreadable] DESCRIPTION (provided by applicant): Following vesicle fusion and exocytosis of neurotransmitter, vesicular retrieval is critical to allow transmission to continue during extended activity. The demands on the vesicle cycle are particularly great in tiny nerve terminals that have a limited number of vesicles. Classical full-collapse fusion (FCF) proceeds by vesicles completely flattening into the plasma membrane, thereby releasing their transmitter, but also completely losing their lipid and protein content and spherical shape. Vesicles are then reconstructed via clathrin- mediated membrane retrieval. Another set of phenomena generically known as "kiss and run" (K&R) has been proposed wherein vesicular identity is maintained and the same vesicle is repeatedly reused. The likely consequence is increased synaptic efficiency and information throughput. To gain further insights into these apparently distinct fusion modes, the following specific aims are proposed: (1) To describe fundamental properties of modes of fusion/retrieval using novel optical approaches. New measurements will indicate the time course by which vesicles undergo fusion for the first time after a period of rest and provide a simple way of dissecting the contributions of FCF and K&R. Quantum dots (QDs) will be used to specifically register FCF events. Modification of QDs by conjugation with the pH-sensitive dye Flubi2 will permit real-time, single vesicle tracking of both K&R and FCF. (2) To clarify key structural determinants of fusion/retrieval modes. The new techniques will allow examination of the cell biological and molecular factors that govern the balance between K&R and FCF. Do vesicles in the readily releasable pool (RRP) continue to stay in the RRP over multiple bouts of release? Can the concept of vesicle reuse be demonstrated definitively? The location of RRP and total recycling pool (TRP) vesicles will be examined. Candidate molecular mechanisms for regulation of fusion/retrieval modes involving dynamin-1 and -2 will be tested. (3) To determine the impact of fusion/retrieval modes on quanta! neurotransmission. Single quantal events during K&R and FCF will be separately analyzed by recording excitatory postsynaptic currents in combination with imaging of fusion/retrieval events. This work will clarify how vesicles are deployed for efficient transmission in nerve terminals and how this usage is modified by parameters such as resting potential and firing pattern. Such questions are central to understanding synaptic communication in both healthy and diseased neural circuits. [unreadable] [unreadable]

The overall goal of this project is to gain a clearer picture of how voltage-gated calcium channels link membrane excitation to rises in intracellular Ca2+ and vital cellular responses. The specific focus is on the alpha1A subunit, which generates P/Q type channels, the dominant pathway for Ca2+ current in many CNS neuron cell bodies and nerve terminals. Deletion of alpha1A drastically changes the profile of pre- and postsynaptic Ca2+ entry and alters key properties of synaptic transmission; mutations in alpha1A can cause neurological diseases, including migraine, episodic ataxia, and spinocerebellar ataxia in human, and various forms of absence epilepsy in mice. Here one major aim is to use molecular strategies to shed light on the molecular relationship between Ca2+ channels and Ca2+ sensors for transmitter release and short term facilitation. Engineered versions of human or mouse alpha1A subunits will be introduced as transgenes driven by the endogenous mouse promoter. Among the modified alpha1A subunits will be one unable to support Ca2+ permeation but with cytoplasmic 0domains intact, and another fully able to permeate but lacking in domains thought to be necessary for interaction with the release machinery. In this way, distinctions will be made between possible roles of alpha1A subunits as providers of Ca2+ influx and a putative structural elements within the synapse. Similar approaches will be directed toward understanding familial hemiplegic migraine (FHM), which arises from points mutations in alpha1A. Previous biophysical studies of the FHM mutants expressed in oocytes or HEK293 cells have not yielded a clear picture of what Ca2+ channel characteristics cause the disease. However, it is expected that a more consistent pattern will emerge when these mutants are studies in a physiological setting within mammalian brain cells and their participation in Ca2+ homeostasis is taken into account. Regulation of pre- and postsynaptic Ca2+ transients and synaptic transmission will be examined to see how these are affected by the various modifications in alpha1A. This work may provide useful clues about the dominant pattern of inheritance of FHM and the intermittent features of the disease.

DESCRIPTION (provided by applicant): Synaptic strength must be regulated in the face of changing levels of input in order to ensure that total strength of all inputs are controlled so as to maintain output firing within reasonable limits and these mechanisms must work in conjunction with mechanisms of synaptic plasticity. Its clinical importance can be best illustrated by the condition of epilepsy where these mechanisms are disrupted at the extreme. The association of epilepsy with numerous forms of inherited disorders of cognitive function such as Fragile X and Down Syndrome and the increasing use of anti-epileptic drugs for the treatment of mood disorders highlight the importance of these mechanisms in maintaining normal brain function. This grant proposes to study the underlying mechanisms by which synaptic adaptation is achieved in the face of changing levels of input and the effects of this adaptation on plasticity, building on recent progress in the lab. The first two parts will use strategies to describe more completely the signaling events in the pathway from synaptic input to synaptic modification, taking advantage of the ease of manipulation and measurement in the primary culture system. This part of the study will begin with a careful dissection of the sources of electrical input and calcium entry responsible for the induction of the key signaling events, followed by an elucidation of the mechanisms of regulation of key intermediate signaling products, examining the regulation of their enzymatic activity, stability, transcription, translation and transport to the synapse. In addition, a more complete description of the signaling cascade will be drawn out from microarray studies comparing results from disruption of the signaling pathway at various points. The next part of the study will look closely at the microphysiology of this phenomenon (a) to elucidate the elements of adaptation that are cell-autonomous, by comparing effects of network changes and manipulations in only one cell (b) to describe adaptive changes at the level of single synapses, and (c) to determine the pre- or postsynaptic locus of induction of the various elements of adaptation. These studies will take advantage of long standing expertise in the lab in studying the physiology of single synapses involving focal stimulation and measurement of synaptic responses at unitary sites. The last part of the project will apply the knowledge gained in the previous sections to try to understand the impact of adaptation m intact hippocampal circuits, using the organotypic slice culture. Primary questions addressed will be differences in the expression of adaptation at two different sets of afferents and consequences of this adaptation on the induction and expression of LTP. We will look for forms of LTP not previously found in pyramidal neurons and for changes in the performance of preexisting mechanisms of LTP.

Abstract not provided.

bioimaging /biomedical imaging; technology /technique development

DESCRIPTION (provided by applicant): Optical tools to dissect synaptic changes underlying epilepsy A prevailing hypothesis about epilepsy contends that neural circuits become overexcitable because of a pathological imbalance between synaptic excitation and inhibition. However, many questions remain about whether this is the dominant principle of epileptiform activity, and whether the imbalance comes about because excitatory synapses are bolstered, because inhibitory synapses are weakened, or both. These issues are challenging to approach, in part because conventional recordings of network activity do not allow the strength of individual types of synaptic input to be readily resolved or dissected. To overcome such difficulties, we are engaged in developing new approaches that use genetically encoded optical indicators to track the contributions of different kinds of presynaptic terminal. We have constructed a new optical probe for vesicle fusion, called sypHTomato, which fluoresces in the red when synaptic vesicles fuse and release neurotransmitter. We are currently generating a mouse that will express sypHTomato within specific types of neurons under control of genetically targetable enzyme, Cre recombinase. SypHTomato can be used in conjunction with existing green probes such as synaptopHluorin or GCaMP3. This will enable independent and simultaneous monitoring at multiple types of synapses, be they excitatory, generically inhibitory, or inhibitory neurons of a particular subclass; it will also allow synaptic activity to be tracked along with action potential firing. We will validate and optimize this two-color system, using neural networks of increasing complexity and relevance to epilepsy. Optical recordings will be performed in conjunction with advanced methods for electrical recording currently in use within the lab. Probes for monitoring synaptic activity will be co-expressed in conjunction with light-sensitive proteins such as Channelrhodopsin-2 and Halorhodopsin to allow manipulation of selected synaptic inputs to a circuit while monitoring the output. In this way, the activity of specific synapses can be assessed during the development of interictal and ictal activity in brain slices and that activity can be further enhanced or turned off by appropriate illumination as tests of their causative role. As proof-of-principle, we will clarify the changes in synaptic input that favor or restrain the genesis of epileptiform bursts in select experimental models of epilepsy. Our molecular reagents, animals and technical approaches will be freely available to epilepsy investigators and to the scientific community at large. The reporter strategy can be easily integrated with existing lines of mice that serve as animal models of human epilepsy. Thus, powerful optical approaches to elucidate the underpinnings of epileptiform activity can be readily put to use in a wide range of mutational and experimental settings, thereby leveraging recent advances in the genetics of epilepsy.

ABSTRACT Homeostatic regulation of excitability and synaptic efficacy works in conjunction with acutely induced Hebbian plasticity to maintain neuron firing within limits and thus preserve network stability and information flow. There is general agreement that homeostatic plasticity can affect intrinsic properties (action potential duration controlling neurotransmission) or synaptic properties (unitary synaptic current amplitude, for example) and involves diverse molecular mechanisms. Dysfunctional homeostasis has been invoked as a basis for brain diseases such as autism spectrum disorders (ASD). Despite major effort, the molecular underpinnings of various forms of homeostatic adaptation are still not clear. In this project, we will examine various aspects of neuronal homeostasis with relevance to neuropsychiatric disorders. The first question is how neuronal inactivity initiates local signaling near postsynaptic CaV1 channels and causes propagation of signals to the nucleus to regulate alternative mRNA splicing (AS) and thus affect spike duration. We will extend our findings on how one ASD-related gene (CACNA1C, L-type Ca2+ channel subunit) controls the expression of another (KCNMA1, BK channel subunit). Our data suggest that signaling to the nucleus via bCaMKK (encoded by CAMKK2) plays a critical role in AS through effects on localization of the splice factor Nova-2. In another subproject, we will clarify how the same activity silencing affects synaptic properties, and the striking switchover of postsynaptic glutamate receptors from Ca2+-impermeable to Ca2+-permeable AMPA receptors. We will decipher how various signaling pathways, generating both negative and positive feedback, work in coordination to trigger a damped oscillatory response of synaptic properties following TTX silencing, a novel observation from our group. We will take studies of homeostasis to recurrent circuits in cultured hippocampal slices, using an all-optical approach to visualize reallocation of presynaptic weights following inactivity and their postsynaptic consequences. Each of the Aims are of relevance to disease states such as ASD and schizophrenia. Using a mouse model of Timothy Syndrome, a rare form of ASD, we will probe how physiological phenomena are altered in a pathogenic setting, for example exploring why inactivity-driven BK splicing is much more severe in Timothy Syndrome neurons and probing how this affects higher order functions of relevance to ASD.

Oxytocin is a peptide hormone synthesized and released from the hypothalamus for reproduction and maternal behavior. Recent studies have tagged oxytocin as a ?trust? hormone, promising to improve social deficits in various mental disorders, such as autism. Despite the enthusiasm for oxytocin, contradictory results in the efficacy of oxytocin in improving human social behaviors have been reported. Such inconsistency in literature is likely due to our poor understanding of complexity of oxytocin action, which likely varies with behavioral state, experience and brain structures. We believe that a better understanding of the endogenous action of oxytocin is the key to unleash the therapeutic potential of this highly evolutionary conserved neuropeptide. Advancing our understanding requires cross-level and comparative inter-disciplinary studies by a group of investigators with overlapping interests and the technical capability to analyze oxytocin signaling across molecular, physiological, systems behavioral and levels. This includes multi-animal interactions, as many mental disorders are impactful on social behavior, over the lifespan and throughout the brain. Oxytocin action in maternal brain is especially important as it represents the most ancient and important function of oxytocin under a social context. To these ends, the proposed Brain Initiative Project in NYU School of Medicine on ?Oxytocin Modulation of Neural Circuit Function and Behavior? consists of four inter-related Projects and four Core facilities, including an Administrative Core, a Data Science Core, a Behavioral Optogenetics Core, and an Oxytocin Receptor Antibody Production Core. The overarching goal of the four Projects and the Cores is to achieve a better understanding of the oxytocin modulation in socio-spatial behaviors, which we define as social interactions within a specific context or behavioral environment. Our team will join forces to tackle the oxytocin system from both the source (oxytocin neurons) and the receiving ends (oxytocin receptor-expressing neurons) From the source, Project 1 and 2 will address the connectivity, behavioral influence, in vivo responses, release and experience- dependent changes of the oxytocin neurons. From the receiving ends, Project 3 will dive into detailed cellular, synaptic and microcircuit mechanisms that mediate the oxytocin actions. Lastly, Project 4 will combine the knowledge and techniques developed from Projects 1, 2, and 3 to investigate the state-dependent oxytocin modulation of aggressive behaviors at a brain site essential for aggression, the ventrolateral part of the ventromedial hypothalamus (VMHvl). Intriguingly, a group of oxytocin neurons are found neighboring the VMHvl, potentially providing a local source of oxytocin although its behavioral relevance is currently unknown. Together these Projects and Cores develop new tools, use cutting-edge techniques, and large-scale methods to provide an in-depth description of the neural circuitry for maternal socio-spatial behavior.

Project Summary (Project 3, Co-PIs: Tsien, Froemke, Buzsaki) Neuromodulators act across many timescales?a consequence of the dynamics of their release, receptor activation and downstream signaling. Their actions target numerous subcellular compartments, shaping synaptic transmission, intrinsic excitability and long-term plasticity. How, in turn, these phenomena translate to behavior is a fundamental goal of neuroscience research. In Project 3, we grapple with this complexity by deconstructing the actions of the peptide and hormone oxytocin. Famous for its roles in the periphery and in social behavior, the biophysical and cellular consequences of oxytocin signaling in the central nervous system are poorly described. A thorough understanding of how oxytocin?s role in the brain is further motivated by disruption of oxytocin signaling in various neuropsychiatric disorders, including ASD and schizophrenia. To address this gap in knowledge, we will study the cellular, synaptic and microcircuit signaling mechanisms of oxytocin in the hippocampus, focusing on the CA2 subregion. Long overlooked, CA2 is enriched in OXTRs and, intriguingly, has been implicated in social behavior. Our most recent efforts have focused on how activation of the OXTR depolarizes CA2 pyramidal cells and causes them to enter into a burst firing mode. This effect was attributable to inhibition of a Kv7-mediated potassium current (or M-current), downstream of a Gq- coupled signaling pathway. In Project 3, we take these biophysical results into increasingly more physiological contexts. In Aim 1, we ask how endogenous activity patterns of oxytocinergic fibers translate into oxytocin release, receptor activation and changes in intrinsic excitability. In Aim 2, we test the strength of our model (in which oxytocin?s effects in the hippocampus are primarily mediated by M-current inhibition), by developing optical tools that test the sufficiency and necessity of M-current inhibition in oxytocin signaling. In Aim 3, we ask how profound changes in hippocampal activity, specifically in CA2, are transmitted beyond the hippocampus. We primarily focus our efforts on the lateral septum; a region long implicated in social behaviors, densely innervated by the hippocampus and rich itself in OXTRs. In sum, we propose a research plan that distills oxytocin signaling in the hippocampus into its most elementary components: peptide release, receptor activation and cell-type specific modulation of the M-current. Then, as an acid test of our understanding, we attempt to reconstruct oxytocin?s modulatory actions using our newly developed optical tools. Finally, we consider how oxytocin signaling in the hippocampus may propagate to downstream structures, ultimately influencing social behavior.

Project Summary: Administrative Core This BRAIN Initiative proposal on ?Oxytocin Modulation of Neural Circuit Function and Behavior? will be located at NYU School of Medicine, and each of the four Project team labs (Tsien, Buzsaki, Lin, and Froemke) are housed within the Neuroscience and Skirball Institutes at NYU School of Medicine and NYU Langone Health. This proximity facilitates their interactions, already leading to a number of collaborations that collectively form the foundation for this proposal. Our team studies and activites will therefore benefit from integration with the larger NYU scientific environment, including the Neuroscience and Physiology graduate program, an existing and robust scientific outreach program, and insititional resources including a cutting-edge health science library and data management system together with a world-class biostatistics department. This Administrative Core will help ensure the management, coordination, and interactions between these components of this proposal. Aim 1 of the Administrative Core is to enable productive and synergistic interactions between Project team labs, by organizing the Internal and External Advisory Committees, and scheduling regular meetings for open communication, troubleshooting, and sharing of scientific results. Aim 2 is to enhance the scientific training and careers of junior colleagues in team labs, including graduate students, postdoctoral fellows, and technicians, by co-advising and interactions with NYU training programs. Aim 3 is to ensure that our results are clearly communicated in a timely manner to NIH staff, other scientists, and the public. We will build and maintain a website to feature team members, activities, and discoveries. Our labs are committed to data sharing, and we will work with the proposed Data Science Core on these efforts. The NYU Neuroscience graduate program has substantial connections with high schools and museums throughout New York City, and engages in activities both statewide and national (particularly through the Society for Neuroscience). Finally, Aim 4 will ensure the appropriate management of Project and Core finances and the overall budget. Administrative Core Co-Directors and staff members are highly experienced at managing multi-PI projects and budgets.These aims of the Administrative Core will help maximize Project team productivity, enabling synergistic and accelerated scientific progress across team labs, and effectively communicate our findings to the greater scientific and lay community.