Multimodal imaging for validation and optimization of ion channel-based chemogenetics in nonhuman primates

Chemogenetic tools provide an opportunity to manipulate neuronal activity and behavior selectively and repeatedly in nonhuman primates (NHPs) with minimal invasiveness. Designer Receptors Exclusively Activated by Designer Drugs are one example that is based on mutated muscarinic acetylcholine receptors. Another channel-based chemogenetic system available for neuronal modulation in NHPs uses Pharmacologically Selective Actuator Modules (PSAMs), which are selectively activated by Pharmacologically Selective Effector Molecules (PSEMs). To facilitate the use of the PSAM/PSEM system, the selection and dosage of PSEMs should be validated and optimized for NHPs. To this end, we used a multimodal imaging approach. We virally expressed excitatory PSAM (PSAM4-5HT3) in the striatum and the primary motor cortex of two macaque monkeys, and visualized its location through positron emission tomography (PET) with the reporter ligand [18F]ASEM. Chemogenetic excitability of neurons triggered by two PSEMs (uPSEM817 and uPSEM792) was evaluated using [18F]fluorodeoxyglucose-PET imaging, with uPSEM817 being more efficient than uPSEM792. Pharmacological magnetic resonance imaging showed that increased brain activity in the PSAM4-expressing region began approximately 13 min after uPSEM817 administration and continued for at least 60 min. Our multimodal imaging data provide valuable information regarding the manipulation of neuronal activity using the PSAM/PSEM system in NHPs, facilitating future applications. Significance statement Like other chemogenetic tools, the ion channel-based system called Pharmacologically Selective Actuator Module/Pharmacologically Selective Effector Molecule (PSAM/PSEM) allows remote manipulation of neuronal activity and behavior in living animals. Nevertheless, its application in non-human primates (NHPs) is still limited. Here, we used multi-tracer positron emission tomography (PET) imaging and pharmacological magnetic resonance imaging (MRI) to visualize an excitatory chemogenetic ion channel (PSAM4-5HT3) and validate its chemometric function in macaque monkeys. Our results provide the optimal agonist, dose, and timing for chemogenetic neuronal manipulation, facilitating the use of the PSAM/PSEM system and expanding the flexibility and reliability of circuit manipulation in NHPs in a variety of situations.

[1]  M. Takada,et al.  Chemogenetic attenuation of cortical seizures in nonhuman primates , 2023, Nature Communications.

[2]  Sarah R. Heilbronner,et al.  The use of chemogenetic actuator ligands in nonhuman primate DREADDs-fMRI , 2022, Current research in neurobiology.

[3]  B. Richmond,et al.  Chemogenetic Disconnection between the Orbitofrontal Cortex and the Rostromedial Caudate Nucleus Disrupts Motivational Control of Goal-Directed Action , 2022, The Journal of Neuroscience.

[4]  S. Bouret,et al.  Using Inhibitory DREADDs to Silence LC Neurons in Monkeys , 2022, Brain sciences.

[5]  B. Richmond,et al.  Characterization of ultrapotent chemogenetic ligands for research applications in non-human primates , 2022, bioRxiv.

[6]  P. Rudebeck,et al.  Resting-State fMRI-Based Screening of Deschloroclozapine in Rhesus Macaques Predicts Dosage-Dependent Behavioral Effects , 2021, The Journal of Neuroscience.

[7]  T. Kikusui,et al.  Chemogenetic inactivation reveals the inhibitory control function of the prefronto-striatal pathway in the macaque brain , 2021, Communications Biology.

[8]  I. Fujita,et al.  A mosaic adeno-associated virus vector as a versatile tool that exhibits high levels of transgene expression and neuron specificity in primate brain , 2021, bioRxiv.

[9]  K. Moussawi,et al.  Excitation of medium spiny neurons by ‘inhibitory’ ultrapotent chemogenetics via shifts in chloride reversal potential , 2021, eLife.

[10]  N. Kalin,et al.  Evidence in primates supporting the use of chemogenetics for the treatment of human refractory neuropsychiatric disorders. , 2021, Molecular therapy : the journal of the American Society of Gene Therapy.

[11]  M. Takada,et al.  Chemogenetic dissection of the primate prefronto-subcortical pathways for working memory and decision-making , 2021, Science Advances.

[12]  M. Takada,et al.  Chemogenetic activation of nigrostriatal dopamine neurons in freely moving common marmosets , 2021, bioRxiv.

[13]  B. Richmond,et al.  Single caudate neurons encode temporally discounted value for formulating motivation for action , 2020, bioRxiv.

[14]  I. Hasegawa,et al.  Macaques Exhibit Implicit Gaze Bias Anticipating Others' False-Belief-Driven Actions via Medial Prefrontal Cortex. , 2020, Cell reports.

[15]  M. Baxter,et al.  Effect of chemogenetic actuator drugs on prefrontal cortex-dependent working memory in nonhuman primates , 2020, Neuropsychopharmacology.

[16]  Samuel T. Slocum,et al.  Deschloroclozapine, a potent and selective chemogenetic actuator enables rapid neuronal and behavioral modulations in mice and monkeys , 2019, Nature Neuroscience.

[17]  Ara M. Abramyan,et al.  High-potency ligands for DREADD imaging and activation in rodents and monkeys , 2019, Nature Communications.

[18]  Marleen Verhoye,et al.  Chemogenetic silencing of neurons in the mouse anterior cingulate area modulates neuronal activity and functional connectivity , 2019, NeuroImage.

[19]  M. Michaelides,et al.  Ultrapotent chemogenetics for research and potential clinical applications , 2019, Science.

[20]  Christa Payne,et al.  Chemogenetic Inhibition of the Amygdala Modulates Emotional Behavior Expression in Infant Rhesus Monkeys , 2019, eNeuro.

[21]  Willem M. Otte,et al.  A novel approach to map induced activation of neuronal networks using chemogenetics and functional neuroimaging in rats: A proof-of-concept study on the mesocorticolimbic system , 2017, NeuroImage.

[22]  Yasuyuki Kimura,et al.  PET imaging-guided chemogenetic silencing reveals a critical role of primate rostromedial caudate in reward evaluation , 2016, Nature Communications.

[23]  Chad J. Donahue,et al.  Using Diffusion Tractography to Predict Cortical Connection Strength and Distance: A Quantitative Comparison with Tracers in the Monkey , 2016, The Journal of Neuroscience.

[24]  B. Roth.  DREADDs for Neuroscientists , 2016, Neuron.

[25]  Richard C Saunders,et al.  Chemogenetic disconnection of monkey orbitofrontal and rhinal cortex reversibly disrupts reward value , 2015, Nature Neuroscience.

[26]  Martin G Pomper,et al.  18F-ASEM, a Radiolabeled Antagonist for Imaging the α7-Nicotinic Acetylcholine Receptor with PET , 2014, The Journal of Nuclear Medicine.

[27]  K. Masui,et al.  Evaluation of the predictive performance of a pharmacokinetic model for propofol in Japanese macaques (Macaca fuscata fuscata). , 2013, Journal of veterinary pharmacology and therapeutics.

[28]  L. Looger,et al.  Chemical and Genetic Engineering of Selective Ion Channel–Ligand Interactions , 2011, Science.

[29]  Mark W. Woolrich,et al.  Advances in functional and structural MR image analysis and implementation as FSL , 2004, NeuroImage.