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Principal Investigator: Shana M Augustin
Organization: NORTHWESTERN UNIVERSITY AT CHICAGO
Fiscal Year: 2024
Award: $222,978
Funding agency: National Institute on Alcohol Abuse and Alcoholism
Neuromodulation is crucial for information processing throughout the brain. Neuromodulators influence neuronal function by acting through G protein-coupled receptors (GPCRs) to alter neuronal excitability and synaptic transmission, which can then affect circuit functions. GPCRs are major drug targets used to treat a variety of diseases, including neurological disorders. The causal link between in vivo subcellular signaling mechanisms and behaviors is poorly understood due to the limited tools available to monitor signaling in freely behaving animals. Activation of GPCRs stimulates G-protein signaling to increase or decrease cyclic monophosphate (cAMP) accumulation and bidirectionally control Protein Kinase A (PKA) and Exchange Protein directly Activated by cAMP (EPAC) signaling. Although GPCRs are diverse, the downstream second messenger systems are limited. Therefore, the overarching hypothesis of this proposal is that GPCRs decode incoming modulatory inputs by generating distinct spatiotemporal patterns of cAMP-mediated signaling to control basal ganglia circuit functions. To test this hypothesis, I propose two innovative specific aims: Specific Aim 1 – I will determine the spatiotemporal dynamics in real- time of A-kinase phosphorylation using virally expressed A-kinase activity reporter (AKAR) and cAMP using the EPAC Föster resonance energy transfer (FRET) - based sensors before and after the induction of striatal long- term depression (LTD) in specific cell types. To execute this Aim, I will use transgenic mice to target specific neuronal cell types and two-photon fluorescence lifetime imaging microscopy (FLIM) to quantify FRET activity. These results will build on my previous published findings and will be of broad interest to the basal ganglia field. Specific Aim 2 – I will monitor cAMP and PKA temporal signaling profiles in specific striatal cell types in freely-moving mice during spontaneous locomotion and motor-skill learning on the accelerated rotarod using virally expressed AKAR and EPAC sensors and deep brain in vivo fiber photometry. This proposal will be the first to determine the cAMP mediated signaling dynamics in striatum during synaptic plasticity and learned behaviors. Throughout my career, I have been interested in determining the causal link between synaptic plasticity and behaviors. At every stage of my career, I have advanced in my technical abilities and refined my scientific experimental design. As I train with my mentors, Drs. Lovinger and Vogel, I will further expand my technical abilities and increase my scientific sophistication to ask impactful questions and design appropriate experiments to address these questions. Additionally, my mentors will train me to communicate my scientific findings effectively, run a successful lab, and mentor to students. I have recruited two extramural investigators, Drs. Cheer and Gremel to serve as advisory committee members and aid in my successful transition to an independent faculty position. Together, my mentors will ensure that I am trained in the skills required to attain a tenure-track faculty position and succeed as an independent research investigator.
Terms: <2-photon><3'5'-cyclic ester of AMP><3,5 cyclic AMP synthetase><AC5 enzyme><Acceleration><Address><Adenosine Cyclic 3',5'-Monophosphate><Adenosine Cyclic Monophosphate><Adenosine Cyclic Monophosphate-Dependent Protein Kinases><Adenosine, cyclic 3',5'-(hydrogen phosphate)><Adenyl Cyclase><Adenylate Cyclase><Adenylyl Cyclase><Advisory Committees><Affect><Anatomic Sites><Anatomic structures><Anatomy><Animals><Basal Ganglia><Basal Nuclei><Behavior><Behavioral><Brain><Brain Nervous System><Causality><Cell Body><Cell Communication and Signaling><Cell Culture Techniques><Cell Signaling><Cells><Cognition><Committee Members><Communication><Corpus Striatum><Corpus striatum structure><Cyclic AMP><Cyclic AMP-Dependent Protein Kinases><Cyclicity><Data><Decision Making><Development><Disease><Disorder><Dissociation><Dorsal><Drug Targeting><Drug usage><Drugs><EXTMR><Encephalon><Endocrine Gland Secretion><Energy Transfer><Ensure><Equilibrium><Etiology><Experimental Designs><Extramural><Extramural Activities><FLIM imaging><FRET><Faculty><Fiber><Fiber Optics><Fluorescence Resonance Energy Transfer><Fostering><Förster Resonance Energy Transfer><G Protein-Complex Receptor><G Protein-Coupled Receptor Genes><G-Protein-Coupled Receptors><G-Proteins><GPCR><GTP-Binding Proteins><GTP-Regulatory Proteins><Guanine Nucleotide Coupling Protein><Guanine Nucleotide Regulatory Proteins><Hormones><Image><Imaging Procedures><Imaging Technics><Imaging Techniques><Impairment><Intracellular Communication and Signaling><Intracellular Second Messenger><Investigators><Kinases><Lateral><Learning><Learning Skill><Ligand Binding><Ligands><Link><Lipids><Location><Locomotion><Long-Term Depression><Long-Term Synaptic Depression><Measures><Mediating><Medication><Medicine><Memory><Mentors><Mice><Mice Mammals><Molecular Fingerprinting><Molecular Profiling><Molecular Target><Monitor><Moods><Motor><Murine><Mus><Nerve Cells><Nerve Impulse Transmission><Nerve Transmission><Nerve Unit><Nervous System Diseases><Nervous System Disorder><Neural Cell><Neural Transmission><Neurocyte><Neurologic Disorders><Neurological Disorders><Neuromodulator><Neuronal Transmission><Neurons><Olfaction><Organism-Level Process><Organismal Process><Outcome><PKA><Paralysis Agitans><Parkinson><Parkinson Disease><Pathway interactions><Pattern><Performance><Periodicity><Pharmaceutical Preparations><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Photometry><Photons><Physiologic><Physiologic Processes><Physiological><Physiological Processes><Position><Positioning Attribute><Preparation><Primary Parkinsonism><Process><Protein Kinase A><Protein Phosphorylation><Proteins><Publishing><Receptor Protein><Reporter><Research><Research Personnel><Researchers><Rhythmicity><Running><Second Messenger Systems><Second Messengers><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Signaling Factor Proto-Oncogene><Signaling Pathway Gene><Signaling Protein><Smell><Smell Perception><Specificity><Striate Body><Striatum><Students><Synapses><Synaptic><Synaptic Transmission><Synaptic plasticity><System><Task Forces><Testing><Therapeutic Hormone><Time><Tissue imaging><Training><Transgenic Mice><Transphosphorylases><Viral><adenosine 3'5' monophosphate><adenylyl cyclase type 5><adenylyl cyclase type V><advisory team><axon signaling><axon-glial signaling><axonal signaling><balance><balance function><behavior response><behavioral response><biological signal transduction><brain tissue><cAMP><cAMP-Dependent Protein Kinases><career><causation><cell culture><cell cultures><cell type><design><designing><developmental><disease causation><drug use><drug/agent><experiment><experimental research><experimental study><experiments><fluorescence life-time imaging><fluorescence life-time imaging microscopy><fluorescence lifetime imaging><fluorescence lifetime imaging microscopy><glia signaling><glial signaling><imaging><improved><in vivo><information processing><innovate><innovation><innovative><interest><learned behavior><learning behavior><longterm depression><longterm synaptic depression><molecular profile><molecular signature><motor skill learning><nerve signaling><neural><neural control><neural regulation><neural signaling><neurological disease><neuromodulation><neuromodulatory><neuronal><neuronal excitability><neuronal signaling><neuroregulation><neurotransmission><odor perception><olfactory perception><optic imaging><optical imaging><pathway><perceptual motor learning><postsynaptic><preparations><receptor><recruit><response><sensor><shRNA><short hairpin RNA><signal transduction second messengers><skills><small hairpin RNA><spatiotemporal><striatal><synapse><tenure process><tenure track><tool><two-photon>