Microscopy Using Fluorescent Drug Biosensors for “Inside-Out Pharmacology”

Muthusamy, Anand K.; Shivange, Amol V.; Nichols, Aaron; Kamajaya, Aron; Jeon, Janice; Borden, Philip; Marvin, Jonathan S.; Unger, Elizabeth K.; Bao, Huan; Chapman, Edwin; Tian, Lin; Looger, Loren L.; Lester, Henry A.

doi:10.1016/j.bpj.2017.11.1990

Cited by 2 publications

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“…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

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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%

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

Zhang

Shivange

et al. 2020

Preprint

Self Cite

138

View full text Add to dashboard Cite

show abstract

“…Other neuronal drugs may also operate via inside-out pathways (Jong et al, 2009; Lester et al, 2012, 2015). With further modifications to the ligand site, preliminary data show that it may be possible to develop families of biosensors for optical subcellular pharmacokinetics of several amine-containing drug classes (Muthusamy et al, 2018; Shivange et al, 2017. Annual Meeting of the Society of General Physiologists.…”

Section: Discussionmentioning

confidence: 99%