Designing Caged Compounds for Spatiotemporal Control of Cellular Chemistry

Furuta, Toshiaki

doi:10.5059/yukigoseikyokaishi.70.1164

Cited by 16 publications

(11 citation statements)

References 29 publications

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“…We previously developed Bhc caged compounds of various biologically active molecules that exhibit high photolytic efficiencies 28,45,46,47 . With the aim of expanding the repertoire of Bhc caging groups, we also reported platforms of modular caged compounds that can be modified easily by the introduction of various functional units 32,40,41 .…”

Section: Discussionmentioning

confidence: 99%

Design, Synthesis, and Photochemical Properties of Clickable Caged Compounds

Suzuki

Shiraishi

Aoki

et al. 2019

JoVE

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Caged compounds enable the photo-mediated manipulation of the cell physiology with high spatiotemporal resolution. However, the limited structural diversity of currently available caging groups and the difficulties in synthetic modification without sacrificing their photolysis efficiencies are obstacles to expanding the repertoire of caged compounds for live cell applications. As the chemical modification of coumarin-type photocaging groups is a promising approach for the preparation of caged compounds with diverse physical and chemical properties, we report a method for the synthesis of clickable caged compounds that can be modified easily with various functional units via the copper(I)-catalyzed Huisgen cyclization. The modular platform molecule contains a (6-bromo-7-hydroxycoumarin-4-yl)methyl (Bhc) group as a photo-caging group, which exhibits a high photolysis efficiency compared to those of the conventional 2-nitrobenzyls. General procedures for the preparation of clickable caged compounds containing amines, alcohols, and carboxylates are presented. Additional properties such as the water solubility and cell targeting ability can be readily incorporated into clickable caged compounds. Furthermore, the physical and photochemical properties, including the photolysis quantum yield, were measured and were found to be superior to those of the corresponding Bhc caged compounds. The described protocol could therefore be considered a potential solution for the lack of structural diversity in the available caged compounds.

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Section: Discussionmentioning

confidence: 99%

Design, Synthesis, and Photochemical Properties of Clickable Caged Compounds

Suzuki

Shiraishi

Aoki

et al. 2019

JoVE

Self Cite

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“…Furthermore, the spatial resolution of these compounds may be compromised due to the rapid diffusion of the caged compounds. At this point, it is worth mentioning that the application of the photocaging techniques in whole bodies of animals is still challenging and restricted to transparent model organisms such as zebrafish embryos [ 202 ]. The irreversible photoreactions, the low cellular delivery capability, the high concentration requirement, and the limited penetration within the deep tissue are the prominent obstacles for in vivo applications [ 121 , 123 , 203 ].…”

Section: Optogenetics To Unravel the Mechanism Of Ca 2+ Ion Channelsmentioning

confidence: 99%

Deciphering Molecular Mechanisms and Intervening in Physiological and Pathophysiological Processes of Ca2+ Signaling Mechanisms Using Optogenetic Tools

Maltan

Najjar

Tiffner

et al. 2021

Cells

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Calcium ion channels are involved in numerous biological functions such as lymphocyte activation, muscle contraction, neurotransmission, excitation, hormone secretion, gene expression, cell migration, memory, and aging. Therefore, their dysfunction can lead to a wide range of cellular abnormalities and, subsequently, to diseases. To date various conventional techniques have provided valuable insights into the roles of Ca2+ signaling. However, their limited spatiotemporal resolution and lack of reversibility pose significant obstacles in the detailed understanding of the structure–function relationship of ion channels. These drawbacks could be partially overcome by the use of optogenetics, which allows for the remote and well-defined manipulation of Ca2+-signaling. Here, we review the various optogenetic tools that have been used to achieve precise control over different Ca2+-permeable ion channels and receptors and associated downstream signaling cascades. We highlight the achievements of optogenetics as well as the still-open questions regarding the resolution of ion channel working mechanisms. In addition, we summarize the successes of optogenetics in manipulating many Ca2+-dependent biological processes both in vitro and in vivo. In summary, optogenetics has significantly advanced our understanding of Ca2+ signaling proteins and the used tools provide an essential basis for potential future therapeutic application.

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“…One such approach, caged compound photoactivation, allows the application of biologically active molecules with micrometer spatial and sub-second temporal resolution. When treated with light of sufficient energy, the photoremovable protecting group on a caged compound separates from the biologically active form of the molecule. − This photochemical delivery method has been used to activate biologically dormant or inactive biosystems such as neurons and sensory cells in ex vivo and in vivo tissue platforms as well as reactions at the molecular level such as enzymatic reactions and protein folding. Moreover, the protecting group’s rapid release occurs within nanoseconds (ns) to microseconds (μs) depending on the protecting group and the substrate, making this photoactivation process compatible for probing temporal and spatial parameters by electrophysiological and electrochemical detection methods.…”

mentioning

confidence: 99%

“…Moreover, the protecting group’s rapid release occurs within nanoseconds (ns) to microseconds (μs) depending on the protecting group and the substrate, making this photoactivation process compatible for probing temporal and spatial parameters by electrophysiological and electrochemical detection methods. For example, caged substrate photoactivation combines the photorelease process with patch clamp detection to examine the influence of the amino acids glutamate and γ-aminobutyric acid (GABA) as agonists and antagonists for neuronal function. ,,,− The function of dopaminergic neurons and circuits can be explored using recently developed caged dopamine and its caged receptor antagonists. , …”

mentioning

confidence: 99%

In Situ Electrochemical Monitoring of Caged Compound Photochemistry: An Internal Actinometer for Substrate Release

et al. 2021

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Caged compounds are molecules that release a protective substrate to free a biologically active substrate upon treatment with light of sufficient energy and duration. A notable limitation of this approach is difficulty in determining the degree of photoactivation in tissues or opaque solutions because light reaching the desired location is obstructed. Here, we have addressed this issue by developing an in situ electrochemical method in which the amount of caged molecule photorelease is determined by fastscan cyclic voltammetry (FSCV) at carbon-fiber microelectrodes. Using p-hydroxyphenyl glutamate (pHP-Glu) as our model system, we generated a linear calibration curve for oxidation of 4hydroxyphenylacetic acid (4HPAA), the group from which the glutamate molecule leaves, up to a concentration of 1000 μM. Moreover, we are able to correct for the presence of residual pHP-Glu in solution as well as the light artifact that is produced. A corrected calibration curve was constructed by photoactivation of pHP-Glu in a 3 μL photoreaction vessel and subsequent analysis by high-performance liquid chromatography. This approach has yielded a linear relationship between 4HPAA concentration and oxidation current, allowing the determination of released glutamate independent of the amount of light reaching the chromophore. Moreover, we have successfully validated the newly developed method by in situ measurement in a whole, intact zebrafish brain. This work demonstrates for the first time the in situ electrochemical monitoring of caged compound photochemistry in brain tissue with FSCV, thus facilitating analyses of neuronal function.

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Designing Caged Compounds for Spatiotemporal Control of Cellular Chemistry

Cited by 16 publications

References 29 publications

Design, Synthesis, and Photochemical Properties of Clickable Caged Compounds

Design, Synthesis, and Photochemical Properties of Clickable Caged Compounds

Deciphering Molecular Mechanisms and Intervening in Physiological and Pathophysiological Processes of Ca2+ Signaling Mechanisms Using Optogenetic Tools

In Situ Electrochemical Monitoring of Caged Compound Photochemistry: An Internal Actinometer for Substrate Release

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