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Principal Investigator: HUI-CHEN LU
Organization: TRUSTEES OF INDIANA UNIVERSITY
Fiscal Year: 2024
Award: $451,319
Funding agency: National Institute on Drug Abuse
SPECIFIC AIMS-Multiscale Imaging Core (MSIC)
Addictive substances trigger plasticity at the molecular, cellular and circuit levels that manifest as persistent
behavioral changes that may cause substance use disorders. Targeting these changes may lead to novel
strategies for preventing or treating substance use disorders. However, our knowledge of the molecular changes,
the cellular processes and the abnormal circuit activity patterns that underlie various aspects of substance use
disorders including compulsion, loss of intake control, withdrawal, and relapse is rather limited. To facilitate a
better understanding of the molecular to circuit level plasticity accompanying drug abuse, the C3A multi-
scale imaging core will support center investigators, affiliates from the Midwest and beyond, and trainees at
different career stages to acquire the conceptual and technical know-how, and to access state-of-the-art
equipment for nanoscale molecular measurements, for microscale anatomical analysis of subcellular and
cellular profiles and for mesoscale physiological imaging of brain circuits. The C3A multi-scale imaging core
will provide unprecedented imaging opportunities to examine models of substance use disorders at multiple
levels, including: (1) molecular and cellular level imaging with internationally unique cell-type- and subcellular
compartment-specific correlated STORM super-resolution imaging, and its recently developed
PharmacoSTORM extension for nanoscale pharmacology; (2) circuit level 2P imaging to examine selective
neural circuits and cell-type-specific dynamic physiological changes among large cell populations.
Aim 1. Determine the cell- and subcellular compartment-specific nanoscale molecular and microscale
cellular alterations triggered by chronic exposure to drugs of abuse. By employing fluorescent small
molecule-based PharmacoSTORM single-molecule nanoscale pharmacology and antibody-based
ImmunoSTORM super-resolution imaging, we and C3A-affiliated researchers will determine if chronic drug
exposure and/or withdrawal elicit persistently altered nanoscale distribution and abundance of important
signaling proteins in the cell types and brain circuits that are most relevant for substance use disorders. By
correlating the nanoscale molecular measurements with microscale confocal microscopy data, we will also
establish the associated morphological changes in identified subcellular compartments. Particular attention will
be devoted to CB1 cannabinoid and D3 dopamine receptors that have essential roles in all phases of the addiction
cycle and whose antagonists/negative allosteric modulators are among NIDA’s ten highest medication
development priorities.
Aim 2. Characterize the mesoscale circuit rewiring of long-range glutamatergic, dopaminergic and
serotonergic axons induced by developmental or chronic exposure to drugs of abuse. Axon tracts
connecting distant brain regions follow irregular trajectories, thus white matter morphology is difficult to evaluate
by standard brain section staining. Therefore, we will exploit our experience in ScaleS methodology combined
with optimized 2P imaging of the entire mouse brain. This approach will be used to determine the impact of
developmental exposure to THC and other drugs on the integrity and trajectory of identified long-range axons.
Because prenatal cannabis exposure modifies human neural circuits and rodent studies found that developing
long-range glutamatergic axons are particularly sensitive to THC, we will initially determine the impact of perinatal
THC exposure on glutamatergic axons originating from medial prefrontal cortex to various brain regions.
Aim 3. Use in vitro and in vivo 2P sensor imaging to determine the mesoscale physiological changes in
brain circuits elicited by chronic exposure to drugs of abuse. Recent advances in genetically encoded
sensors for Ca2+, endocannabinoids, and monoamines provide excellent tools to visualize dynamic changes of
these signaling molecules in a specific cell-type-specific manner in real-time. By combining our established and
comprehensive methodology for Ca2+_imaging in acute brain slices or awake behaving mice (as young as ten
days old) extending from the surgical procedure through the data analysis pipeline with High Performance
Computing together with GRAB-eCB2.1 and GRABDA sensor imaging, we will support center and affiliated
scientists to perform longitudinal 2P imaging to examine endocannabinoid, dopamine, and network activity
changes in their relevant models of substance use disorders. We will also determine if perinatal THC exposure
perturbs the development of endocannabinoid signaling in association with Ca2+-spike patterns in the primary
somatosensory cortex of awake behaving mouse pups from early postnatal to weaning ages.
Aim 4. Develop in vivo protocols for Fluorescence Lifetime Imaging Microscopy (FLIM) in addiction
research. Drugs of abuse evoke substantial metabolic changes and perturb astrocyte-neuron interactions. We
will use 2P-FLIM imaging to develop in vivo applications using FLIM-based sensors to monitor energy
metabolism, signaling cascades, protein-protein interactions and to estimate the proximity between astrocytes
and neurons in the substance use disorder models established by local and affiliate researchers of the imaging
core.
Terms: <Acute><Age><Anatomic Sites><Anatomic structures><Anatomy><Antibodies><Astrocytes><Astrocytus><Astroglia><Attention><Axon><Behavioral><Brain><Brain Nervous System><Brain imaging><Brain region><CB1><CB1 Receptor><CB1R><CNR1 gene><Cannabinoid Receptor CB1><Cannabinoids><Cannabis><Cell Body><Cell Communication and Signaling><Cell Function><Cell Physiology><Cell Process><Cell Signaling><Cells><Cellular Function><Cellular Physiology><Cellular Process><Chronic><Confocal Microscopy><Data><Development><Distant><Dopamine><Drug Exposure><Drug abuse><Drugs><ECB signaling><Encephalon><Endocannabinoids><Endogenous Cannabinoids><Energy Expenditure><Energy Metabolism><Equipment><Exposure to><FLIM imaging><Functional Imaging><Glutamates><High Performance Computing><Human><Hydroxytyramine><Image><In Vitro><Indiana><Intake><International><Intracellular Communication and Signaling><Investigators><Knowledge><L-Glutamate><Measurement><Medial><Medication><Metabolic><Methodology><Mice><Mice Mammals><Midwest><Midwest U.S.><Midwest US><Midwestern United States><Modeling><Modern Man><Molecular><Monitor><Morphology><Murine><Mus><NIDA><National Institute of Drug Abuse><National Institute on Drug Abuse><Nerve Cells><Nerve Unit><Neural Cell><Neurocyte><Neurons><Operative Procedures><Operative Surgical Procedures><Pattern><Perinatal><Peripartum><Pharmaceutical Preparations><Pharmacology><Phase><Physiologic><Physiologic Imaging><Physiological><Population><Prefrontal Cortex><Protocol><Protocols documentation><Relapse><Research><Research Personnel><Researchers><Rodent><Rodentia><Rodents Mammals><Role><Scientist><Signal Transduction><Signal Transduction Systems><Signaling><Signaling Factor Proto-Oncogene><Signaling Molecule><Signaling Pathway Gene><Signaling Protein><Slice><Somatosensory Cortex><Staining method><Stains><Subcellular Process><Substance Use Disorder><Surgical><Surgical Interventions><Surgical Procedure><THC exposure><Time><Universities><Visualization><Weaning><Withdrawal><abuse of drugs><abused drug><abused drugs><abuses drugs><addiction><addictive disorder><ages><antagonism><antagonist><astrocytic glia><awake><biological signal transduction><brain visualization><cannabinoid receptor 1><cannabinoid receptor type 1><cannabinoid type 1><cannabis in utero><career><cell type><compulsion><data analysis pipeline><data processing pipeline><developmental><disease model><disorder model><dopamine D3 receptor><drug abused><drug of abuse><drug withdrawal><drug/agent><drugs abused><drugs of abuse><endocannabinoid signaling><experience><fetal cannabis exposure><fetal marijuana exposure><fluorescence life-time imaging><fluorescence life-time imaging microscopy><fluorescence lifetime imaging><fluorescence lifetime imaging microscopy><gestational marijuana exposure><glutamatergic><high-end computing><imaging><in vivo><microscope imaging><microscopic imaging><microscopy imaging><molecular scale><monoamine><nano meter scale><nano meter sized><nanometer scale><nanometer sized><nanoscale><neural circuit><neural circuitry><neurocircuitry><neuronal><new approaches><novel approaches><novel strategies><novel strategy><physiological imaging><postnatal><prenatal cannabis exposure><prenatal marijuana exposure><prenatally exposed to cannabis><prenatally exposed to marijuana><prenatally marijuana exposed><prevent><preventing><protein protein interaction><pup><sensor><single molecule><small molecule><social role><somesthetic sensory cortex><substance use and disorder><substantia alba><superresolution imaging><surgery><synaptic circuit><synaptic circuitry><tetrahydrocannabinol exposure><tool><white matter>