Determining the pharmacokinetics of nicotinic drugs in the endoplasmic reticulum using biosensors

Shivange, Amol V.; Borden, Philip; Muthusamy, Anand K.; Nichols, Aaron; Bera, Kallol; Bao, Huan; Bishara, Isaac; Jeon, Janice; Mulcahy, Matthew J.; Cohen, Bruce; O'Riordan, Saidhbhe L.; Kim, Charlene; Dougherty, Dennis A.; Chapman, Edwin; Marvin, Jonathan S.; Looger, Loren L.; Lester, Henry A.

doi:10.1085/jgp.201812201

Cited by 52 publications

(149 citation statements)

References 78 publications

(123 reference statements)

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“…We have developed iAChSnFR into a sensor specific for serotonin by machine learning-guided protein redesign and directed evolution 86 . 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 .…”

Section: Discussionmentioning

confidence: 99%

“…Fig. S5b) and differs by only a few amino acids from the simultaneously developed nicotine sensor, iNicSnFR 36 . For control experiments, we made a non-binding iAChSnFR mutant (iAChSnFR-NULL; mutation Tyr357Ala; no response > 100 µM ACh, choline Kd > 50 mM; Supp.…”

Section: Sensor Engineeringmentioning

confidence: 99%

“…X-ray datasets were collected at Stanford Synchrotron Radiation Lightsource (SSRL) beamline 12-2 with a Pilatus 6M detector using a Blu-Ice interface 106 The data was integrated with XDS 107 , and scaled with aimless 103 . Molecular replacement was carried out using Phaser in Phenix 105 with sub-domains of iNicSnFR (PDB ID: 6EFR) 36 as the input model. Subsequent refinement and model building cycles were carried out with phenix.refine 105 and Coot 104 , respectively.…”

Section: Iachsnfr Periplasmic Binding Protein-onlymentioning

confidence: 99%

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A fast genetically encoded fluorescent sensor for faithful in vivo acetylcholine detection in mice, fish, worms and flies

Zhang

Shivange

et al. 2020

Preprint

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133

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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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Section: Discussionmentioning

confidence: 99%

Section: Sensor Engineeringmentioning

confidence: 99%

Section: Iachsnfr Periplasmic Binding Protein-onlymentioning

confidence: 99%

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A fast genetically encoded fluorescent sensor for faithful in vivo acetylcholine detection in mice, fish, worms and flies

Zhang

Shivange

et al. 2020

Preprint

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133

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“…Intensity-based nicotine-sensing fluorescent reporters (iNicSnFRs): A fluorescent nicotine sensor Determining the relative importance of outside-in and insideout nicotine signaling was previously impossible because the nicotine concentration inside the ER was unknown. To address this knowledge gap, Shivange et al (2019) developed a novel set of engineered protein-based fluorescent sensors called iNicSnFRs. Because the protein-based sensors can be targeted to different subcellular locations, they can provide detailed information about nicotine concentration and dynamics in different organelles.…”

mentioning

confidence: 99%

Inhale, exhale: Probing the inside-out mechanism of nicotine addiction using novel fluorescent sensors

McManus

Werley

Dempsey

2019

Journal of General Physiology

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“…Nicotine crosses the blood brain barrier and binds with high affinity to nAChRs widely distributed throughout the nervous system [ 32 , 38 , 48 , 49 , 50 ]. This interaction promotes a variety of neurophysiological changes including chaperone-mediated nAChR upregulation [ 30 , 35 , 37 , 51 , 52 , 53 , 54 ], activation of the mesocorticolimbic reward and reinforcement pathways [ 55 , 56 ], enhanced synaptic plasticity [ 57 , 58 , 59 , 60 ], and enhanced neuronal firing [ 53 , 60 , 61 , 62 , 63 ], ultimately leading to the development of nicotine addiction [ 33 , 64 ] ( Table 1 ).…”

Section: Background Of Nicotine Addictionmentioning

confidence: 99%

The Impact of Electronic Nicotine Delivery System (ENDS) Flavors on Nicotinic Acetylcholine Receptors and Nicotine Addiction-Related Behaviors

Cooper

Henderson

2020

Molecules

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Over the past two decades, combustible cigarette smoking has slowly declined by nearly 11% in America; however, the use of electronic cigarettes has increased tremendously, including among adolescents. While nicotine is the main addictive component of tobacco products and a primary concern in electronic cigarettes, this is not the only constituent of concern. There is a growing market of flavored products and a growing use of zero-nicotine e-liquids among electronic cigarette users. Accordingly, there are few studies that examine the impact of flavors on health and behavior. Menthol has been studied most extensively due to its lone exception in combustible cigarettes. Thus, there is a broad understanding of the neurobiological effects that menthol plus nicotine has on the brain including enhancing nicotine reward, altering nicotinic acetylcholine receptor number and function, and altering midbrain neuron excitability. Although flavors other than menthol were banned from combustible cigarettes, over 15,000 flavorants are available for use in electronic cigarettes. This review seeks to summarize the current knowledge on nicotine addiction and the various brain regions and nicotinic acetylcholine receptor subtypes involved, as well as describe the most recent findings regarding menthol and green apple flavorants, and their roles in nicotine addiction and vaping-related behaviors.

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Determining the pharmacokinetics of nicotinic drugs in the endoplasmic reticulum using biosensors

Cited by 52 publications

References 78 publications

A fast genetically encoded fluorescent sensor for faithful in vivo acetylcholine detection in mice, fish, worms and flies

A fast genetically encoded fluorescent sensor for faithful in vivo acetylcholine detection in mice, fish, worms and flies

Inhale, exhale: Probing the inside-out mechanism of nicotine addiction using novel fluorescent sensors

The Impact of Electronic Nicotine Delivery System (ENDS) Flavors on Nicotinic Acetylcholine Receptors and Nicotine Addiction-Related Behaviors

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