Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum

Bera, Kallol; Kamajaya, Aron; Shivange, Amol V.; Muthusamy, Anand K.; Nichols, Aaron; Borden, Philip; Grant, Stephen; Jeon, Janice; Lin, Elaine; Bishara, Isaac; Chin, Theodore M.; Cohen, Bruce; Kim, Charlene H.; Unger, Elizabeth K.; Tian, Lin; Marvin, Jonathan S.; Looger, Loren L.; Lester, Henry A.

doi:10.3389/fncel.2019.00499

Cited by 16 publications

(36 citation statements)

References 54 publications

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“…[𝑙𝑖𝑔𝑎𝑛𝑑] at the beginning of the doseresponse relation, emphasizes the response to ligand concentrations in the pharmacologically relevant range (Bera, et al, 2019). Table 1 and Fig.…”

Section: Generation Of Additional Nicotinic Idrugsnfrs: Mutational Tacticmentioning

confidence: 99%

Fluorescence activation mechanism and imaging of drug permeation with new sensors for smoking-cessation ligands

Nichols

Blumenfeld

Fan

et al. 2022

eLife

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Nicotinic partial agonists provide an accepted aid for smoking cessation and thus contribute to decreasing tobacco-related disease. Improved drugs constitute a continued area of study. However, there remains no reductionist method to examine the cellular and subcellular pharmacokinetic properties of these compounds in living cells. Here, we developed new intensity-based drug sensing fluorescent reporters ('iDrugSnFRs') for the nicotinic partial agonists dianicline, cytisine, and two cytisine derivatives - 10-fluorocytisine and 9-bromo-10-ethylcytisine. We report the first atomic-scale structures of liganded periplasmic binding protein-based biosensors, accelerating development of iDrugSnFRs and also explaining the activation mechanism. The nicotinic iDrugSnFRs detect their drug partners in solution, as well as at the plasma membrane (PM) and in the endoplasmic reticulum (ER) of cell lines and mouse hippocampal neurons. At the PM, the speed of solution changes limits the growth and decay rates of the fluorescence response in almost all cases. In contrast, we found that rates of membrane crossing differ among these nicotinic drugs by > 30 fold. The new nicotinic iDrugSnFRs provide insight into the real-time pharmacokinetic properties of nicotinic agonists and provide a methodology whereby iDrugSnFRs can inform both pharmaceutical neuroscience and addiction neuroscience.

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Section: Generation Of Additional Nicotinic Idrugsnfrs: Mutational Tacticmentioning

confidence: 99%

Fluorescence activation mechanism and imaging of drug permeation with new sensors for smoking-cessation ligands

Nichols

Blumenfeld

Fan

et al. 2022

eLife

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show abstract

“…In parallel with development of iAChSnFR, we also mutated OpuBC to recognize nicotine or the smoking-cessation drug varenicline 36 (PDB 6EFR). Further experiments produced variants that detect other neural drugs, including the rapidly acting antidepressant S-ketamine 87 , selective serotonin reuptake inhibitor antidepressants, and opioids 88 . In the original OpuBC and related PBPs 89 , and in the better characterized sensors, ligand binding is mediated by cation-π interactions between the protonated secondary, protonated tertiary, or quaternary amine of the ligand and aromatic residues.…”

Section: Discussionmentioning

confidence: 99%

A fast genetically encoded fluorescent sensor for faithfulin vivoacetylcholine detection in mice, fish, worms and flies

Zhang

Shivange

et al. 2020

Preprint

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Here we design and optimize a genetically encoded fluorescent indicator, iAChSnFR, for the ubiquitous neurotransmitter acetylcholine, based on a bacterial periplasmic binding protein. iAChSnFR shows large fluorescence changes, rapid rise and decay kinetics, and insensitivity to most cholinergic drugs. iAChSnFR revealed large transients in a variety of slice and in vivo preparations in mouse, fish, fly and worm. iAChSnFR will be useful for the study of acetylcholine in all animals. IntroductionAcetylcholine (ACh) is a critical neurotransmitter in all animals. Among invertebrates, it is the most prevalent excitatory transmitter in the brain, sensory ganglia, and frequently the neuromuscular junction (NMJ). Among vertebrates, only a minority of neurons release ACh, but these signals play varying key roles. For instance, ACh signals at the NMJ, in the autonomic nervous system, and in subsets of the central nervous system, particularly projections arising from the brainstem and basal forebrain. Other cholinergic neuron populations in the brain include striatal interneurons, the stria vascularis-medial habenula-interpeduncular nucleus pathway, and sparse, incompletely characterized cell types such as intrinsic cholinergic interneurons in cortex 1 and hippocampus 2 . ACh helps to regulate attention 3 and wakefulness 4 , and participates in memory formation and consolidation 5 . ACh is also an important transmitter in glia, and between the nervous and immune systems 6 .Acetylcholine is synthesized pre-synaptically from choline and acetyl-CoA by choline acetyltransferase (ChAT), then packaged into synaptic vesicles by the vesicular acetylcholine transporter (VAChT). A key, partially understood aspect of cholinergic signaling is co-release with other neurotransmitters, including GABA, ATP, and glutamate 7,8 . To understand the role of co-release, one must measure ACh release alongside emerging measurements of other neurotransmitters.Acetylcholine receptors are among the most diverse neurotransmitter receptor families. Humans possess five muscarinic G protein-coupled receptors (GPCRs) for ACh (mAChRs) with diverse expression in the brain and smooth, cardiac, and skeletal muscle. Vertebrate nicotinic ACh receptors (nAChRs) are pentameric ligand-gated cation channels. Humans have a total of 17 nAChR subunit genes, in five classes: 10 a, 4 b, and one each of g, d, and e. nAChRs occur with many subunit combinations 9 , and others may be undiscovered. Invertebrates also have AChgated chloride channels. On neurons, receptors can be localized pre-, post-, and extrasynaptically, often with different isoforms in each place 10

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“…We examined the subcellular pharmacokinetics of the nicotinic agonists in mammalian cell lines and primary mouse hippocampal neurons. The nicotinic iDrugSnFRs were targeted to the plasma membrane (PM) (iDrugSnFR_PM) or the endoplasmic reticulum (ER) (iDrugSnFR_ER) as previously described (Bera, et al, 2019;Shivange, et al, 2019). We then performed a doseresponse experiment using wide-field fluorescence imaging with each iDrugSnFR and its drug partner, sampling a range of concentrations covering a log scale surrounding the EC50 as determined for the purified protein (Fig.…”

Section: Characterization Of Nicotinic Idrugsnfrs In Hela Cells and Primary Mouse Hippocampal Culturementioning

confidence: 99%

“…[𝑙𝑖𝑔𝑎𝑛𝑑] at the beginning of the dose-response relation, emphasizes the response to ligand concentrations in the pharmacologically relevant range (Bera, et al, 2019). Table 1 and Fig.…”

Section: Generation Of Additional Nicotinic Idrugsnfrs: Mutational Tacticmentioning

confidence: 99%

“…as previously described (Bera, et al, 2019;Shivange, et al, 2019). We then performed a doseresponse experiment using wide-field fluorescence imaging with each iDrugSnFR and its drug partner, sampling a range of concentrations covering a log scale surrounding the EC50 as determined for the purified protein (Fig.…”

Section: Characterization Of Nicotinic Idrugsnfrs In Hela Cells and Primary Mouse Hippocampal Culturementioning

confidence: 99%

See 1 more Smart Citation

Fluorescence Activation Mechanism and Imaging of Drug Permeation with New Sensors for Smoking-Cessation Ligands

Nichols

Blumenfeld

Fan

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Nicotinic partial agonists provide a partial aid for smoking cessation and thus contribute to decreasing tobacco-related disease. Improved drugs constitute a continued area of study. However, there remains no reductionist method to examine the cellular and subcellular pharmacokinetic properties of these compounds in living cells. Here, we developed new intensity-based drug sensing fluorescent reporters (iDrugSnFRs) for the nicotinic partial agonists dianicline, cytisine, and two cytisine derivatives – 10-fluorocytisine and 9-bromo-10-ethylcytisine. Development of the series was aided and explained by the first atomic-scale structural studies on liganded periplasmic binding protein-based biosensors. Members of the series detect their drug partners in solution, as well as at the plasma membrane (PM) and in the endoplasmic reticulum (ER) of cell lines and primary mouse hippocampal neurons. At the PM, the speed of solution changes limits the growth and decay rates of the fluorescence response in almost all cases. In contrast, we found that rates of membrane crossing differ among these nicotinic drugs by > 30 fold. The new nicotinic iDrugSnFRs, in combination with previously described nicotine and varenicline sensors, provide insight into the real-time pharmacokinetic properties of nicotinic agonists and provide a methodology whereby iDrugSnFRs can inform both pharmaceutical neuroscience and addiction neuroscience.

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Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum

Cited by 16 publications

References 54 publications

Fluorescence activation mechanism and imaging of drug permeation with new sensors for smoking-cessation ligands

Fluorescence activation mechanism and imaging of drug permeation with new sensors for smoking-cessation ligands

A fast genetically encoded fluorescent sensor for faithfulin vivoacetylcholine detection in mice, fish, worms and flies

Fluorescence Activation Mechanism and Imaging of Drug Permeation with New Sensors for Smoking-Cessation Ligands

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