@article {68411, title = {Characterization of Ultrapotent Chemogenetic Ligands for Research Applications in Nonhuman Primates.}, journal = {ACS Chemical Neuroscience}, volume = {13}, year = {2022}, month = {2022 Nov 02}, pages = {3118-3125}, abstract = {

Chemogenetics is a technique for obtaining selective pharmacological control over a cell population by expressing an engineered receptor that is selectively activated by an exogenously administered ligand. A promising approach for neuronal modulation involves the use of "Pharmacologically Selective Actuator Modules" (PSAMs); these chemogenetic receptors are selectively activated by ultrapotent "Pharmacologically Selective Effector Molecules" (uPSEMs). To extend the use of PSAM/PSEMs to studies in nonhuman primates, it is necessary to thoroughly characterize the efficacy and safety of these tools. We describe the time course and brain penetrance in rhesus monkeys of two compounds with promising binding specificity and efficacy profiles in studies, uPSEM792 and uPSEM817, after systemic administration. Rhesus monkeys received subcutaneous (s.c.) or intravenous (i.v.) administration of uPSEM817 (0.064 mg/kg) or uPSEM792 (0.87 mg/kg), and plasma and cerebrospinal fluid samples were collected over 48 h. Both compounds exhibited good brain penetrance, relatively slow washout, and negligible conversion to potential metabolites-varenicline or hydroxyvarenicline. In addition, we found that neither of these uPSEMs significantly altered the heart rate or sleep. Our results indicate that both compounds are suitable candidates for neuroscience studies using PSAMs in nonhuman primates.

}, keywords = {Animals, Brain, Ligands, Macaca mulatta, Neurons, Varenicline}, issn = {1948-7193}, doi = {10.1021/acschemneuro.2c00525}, author = {Raper, Jessica and Eldridge, Mark A G and Sternson, Scott M and Shim, Jalene Y and Fomani, Grace P and Richmond, Barry J and Wichmann, Thomas and Galvan, Adriana} } @article {48979, title = {Chemogenetic tools for causal cellular and neuronal biology.}, journal = {Physiological Reviews}, year = {2017}, month = {2017 Dec 12}, abstract = {

Chemogenetic technologies enable selective pharmacological control of specific cell populations. An increasing number of approaches have been developed that modulate different signaling pathways. Selective pharmacological control over G protein-coupled receptor signaling, ion channel conductances, protein association, protein stability, and small molecule targeting allows modulation of cellular processes in distinct cell types. Here, we review these chemogenetic technologies and instances of their applications in complex tissues in vivo and ex vivo.

}, doi = {10.1152/physrev.00009.2017}, author = {Atasoy, Deniz and Sternson, Scott M} } @article {47057, title = {Cell type-specific pharmacology of NMDA receptors using masked MK801.}, journal = {eLife}, volume = {4}, year = {2015}, month = {2015 Sep 11}, abstract = {

N-Methyl-D-aspartate receptors (NMDA-Rs) are ion channels that are important for synaptic plasticity, which is involved in learning and drug addiction. We show enzymatic targeting of an NMDA-R antagonist, MK801, to a molecularly defined neuronal population with the cell-type-selectivity of genetic methods and the temporal control of pharmacology. We find that NMDA-Rs on dopamine neurons are necessary for cocaine-induced synaptic potentiation, demonstrating that cell type-specific pharmacology can be used to dissect signaling pathways within complex brain circuits.

}, issn = {2050-084X}, doi = {10.7554/eLife.10206}, author = {Yang, Yunlei and Lee, Peter and Sternson, Scott M} } @article {47033, title = {Cell type-specific transcriptomics of hypothalamic energy-sensing neuron responses to weight-loss.}, journal = {eLife}, volume = {4}, year = {2015}, month = {2015 Sep 2}, abstract = {

Molecular and cellular processes in neurons are critical for sensing and responding to energy deficit states, such as during weight-loss. AGRP neurons are a key hypothalamic population that is activated during energy deficit and increases appetite and weight-gain. Cell type-specific transcriptomics can be used to identify pathways that counteract weight-loss, and here we report high-quality gene expression profiles of AGRP neurons from well-fed and food-deprived young adult mice. For comparison, we also analyzed POMC neurons, an intermingled population that suppresses appetite and body weight. We find that AGRP neurons are considerably more sensitive to energy deficit than POMC neurons. Furthermore, we identify cell type-specific pathways involving endoplasmic reticulum-stress, circadian signaling, ion channels, neuropeptides, and receptors. Combined with methods to validate and manipulate these pathways, this resource greatly expands molecular insight into neuronal regulation of body weight, and may be useful for devising therapeutic strategies for obesity and eating disorders.

}, issn = {2050-084X}, doi = {10.7554/eLife.09800}, author = {Henry, Fredrick E and Sugino, Ken and Tozer, Adam and Branco, Tiago and Sternson, Scott M} } @article {45167, title = {Chemogenetic synaptic silencing of neural circuits localizes a hypothalamus{\textrightarrow}midbrain pathway for feeding behavior.}, journal = {Neuron}, volume = {82}, year = {2014}, month = {2014 May 21}, pages = {797-808}, abstract = {

Brain function is mediated by neural circuit connectivity, and elucidating the role of connections is aided by techniques to block their output. We developed cell-type-selective, reversible synaptic inhibition tools for mammalian neural circuits by leveraging G protein signaling pathways to suppress synaptic vesicle release. Here, we find that the pharmacologically selective designer Gi-protein-coupled receptor hM4D is a presynaptic silencer in the presence of its cognate ligand clozapine-N-oxide (CNO). Activation of hM4D signaling sharply reduced synaptic release probability and synaptic current amplitude. To demonstrate the utility of this tool for neural circuit perturbations, we developed an axon-selective hM4D-neurexin variant and used spatially targeted intracranial CNO injections to localize circuit connections from the hypothalamus to the midbrain responsible for feeding behavior. This synaptic silencing approach is broadly applicable for cell-type-specific and axon projection-selective functional analysis of diverse neural circuits.

}, keywords = {Agouti-Related Protein, Animals, Basic Helix-Loop-Helix Transcription Factors, Clozapine, Feeding Behavior, Humans, Hypothalamus, Membrane Potentials, Mesencephalon, Mice, Mice, Inbred C57BL, Mice, Transgenic, Models, Molecular, Molecular Sequence Data, Nerve Net, Neural Inhibition, Neurons, Receptor, Muscarinic M4, Repressor Proteins, Rhodopsin, Time Factors}, issn = {1097-4199}, doi = {10.1016/j.neuron.2014.04.008}, author = {Stachniak, Tevye J and Ghosh, Anirvan and Sternson, Scott M} } @article {45051, title = {Chemogenetic tools to interrogate brain functions.}, journal = {Annual Review of Neuroscience}, volume = {37}, year = {2014}, month = {2014 Jul 8}, pages = {387-407}, abstract = {

Elucidating the roles of neuronal cell types for physiology and behavior is essential for understanding brain functions. Perturbation of neuron electrical activity can be used to probe the causal relationship between neuronal cell types and behavior. New genetically encoded neuron perturbation tools have been developed for remotely controlling neuron function using small molecules that activate engineered receptors that can be targeted to cell types using genetic methods. Here we describe recent progress for approaches using genetically engineered receptors that selectively interact with small molecules. Called "chemogenetics," receptors with diverse cellular functions have been developed that facilitate the selective pharmacological control over a diverse range of cell-signaling processes, including electrical activity, for molecularly defined cell types. These tools have revealed remarkably specific behavioral physiological influences for molecularly defined cell types that are often intermingled with populations having different or even opposite functions.

}, issn = {1545-4126}, doi = {10.1146/annurev-neuro-071013-014048}, author = {Sternson, Scott M and Roth, Bryan L} } @article {3977, title = {Chemical and genetic engineering of selective ion channel-ligand interactions.}, journal = {Science}, volume = {333}, year = {2011}, month = {2011 Sep 2}, pages = {1292-6}, abstract = {

Ionic flux mediates essential physiological and behavioral functions in defined cell populations. Cell type-specific activators of diverse ionic conductances are needed for probing these effects. We combined chemistry and protein engineering to enable the systematic creation of a toolbox of ligand-gated ion channels (LGICs) with orthogonal pharmacologic selectivity and divergent functional properties. The LGICs and their small-molecule effectors were able to activate a range of ionic conductances in genetically specified cell types. LGICs constructed for neuronal perturbation could be used to selectively manipulate neuron activity in mammalian brains in vivo. The diversity of ion channel tools accessible from this approach will be useful for examining the relationship between neuronal activity and animal behavior, as well as for cell biological and physiological applications requiring chemical control of ion conductance.

}, keywords = {5-HT3, Animals, Benzamides, Bicyclo Compounds, Brain, Feeding Behavior, Female, Glycine, HEK293 Cells, Humans, Inbred C57BL, Ion Channel Gating, Ligand-Gated Ion Channels, Ligands, Membrane Potentials, Mice, Mutagenesis, Neurons, Nicotinic, Patch-Clamp Techniques, Protein Binding, Protein Engineering, Protein Structure, Quinuclidines, Receptors, Recombinant Fusion Proteins, Serotonin, Small Molecule Libraries, Stereoisomerism, Tertiary}, issn = {1095-9203}, doi = {10.1126/science.1206606}, author = {Magnus, Christopher J and Lee, Peter H and Atasoy, Deniz and Su, Helen H and Looger, Loren L and Sternson, Scott M} }