Highly Zinc-Selective Fluorescent Sensor Molecules Suitable for Biological Applications

Hirano, Tomoya; Kikuchi, Kazuya; Urano, Yasuteru; Higuchi, Tsunehiko; Nagano, Tetsuo

doi:10.1021/ja002467f

Cited by 337 publications

(211 citation statements)

References 27 publications

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“…1). ZIMIR consists of three moieties: a fluorophore based on fluorescein, a Zn 2+ binding motif derived from dipicolylamine (17)(18)(19), and a pair of dodecyl alkyl chains for membrane tethering. In the absence of Zn 2+ , the fluorescence of ZIMIR is quenched by the photo-induced electron transfer from the amino group to fluorescein.…”

Section: Resultsmentioning

confidence: 99%

Imaging dynamic insulin release using a fluorescent zinc indicator for monitoring induced exocytotic release (ZIMIR)

Chen

Bellomo

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

146

176

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Current methods of monitoring insulin secretion lack the required spatial and temporal resolution to adequately map the dynamics of exocytosis of native insulin granules in intact cell populations in three dimensions. Exploiting the fact that insulin granules contain a high level of Zn 2+ , and that Zn 2+ is coreleased with insulin during secretion, we have developed a fluorescent, cell surface-targeted zinc indicator for monitoring induced exocytotic release (ZIMIR). ZIMIR displayed a robust fluorescence enhancement on Zn 2+ chelation and bound Zn 2+ with high selectivity against Ca 2+ and Mg 2+ . When added to cultured β cells or intact pancreatic islets at low micromolar concentrations, ZIMIR labeled cells rapidly, noninvasively, and stably, and it reliably reported changes in Zn 2+ concentration near the sites of granule fusion with high sensitivity that correlated well with membrane capacitance measurement. Fluorescence imaging of ZIMIR-labeled β cells followed the dynamics of exocytotic activity at subcellular resolution, even when using simple epifluorescence microscopy, and located the chief sites of insulin release to intercellular junctions. Moreover, ZIMIR imaging of intact rat islets revealed that Zn 2+ /insulin release occurred largely in small groups of adjacent β cells, with each forming a "secretory unit." Concurrent imaging of ZIMIR and Fura-2 showed that the amplitude of cytosolic Ca 2+ elevation did not necessarily correlate with insulin secretion activity, suggesting that events downstream of Ca 2+ signaling underlie the cell-cell heterogeneity in insulin release. In addition to studying stimulation-secretion coupling in cells with Zn 2+ -containing granules, ZIMIR may find applications in β-cell engineering and screening for molecules regulating insulin secretion on high-throughput platforms.probe development | zinc imaging | hormone secretion assay

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Section: Resultsmentioning

confidence: 99%

Imaging dynamic insulin release using a fluorescent zinc indicator for monitoring induced exocytotic release (ZIMIR)

Chen

Bellomo

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

146

176

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“…11,19 Fluorescence spectroscopy 25 The fluorescence spectra of ZnAF1 and ZnAF2F were recorded at 25 °C on a Cary Eclipse spectrofluorimeter (Varian) in 1 cm cells, using the fluorophore excitation at 492 nm and following its emission in the range of 500-700 nm. Excitation and emission bandwidths of 2.5 nm were used.…”

Section: Introductionmentioning

confidence: 99%

Ternary complex formation and competition quench fluorescence of ZnAF family zinc sensors

2013

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5Our current understanding of the intracellular thermodynamics and kinetics of Zn(II) ions is largely based on the application of fluorescent sensor molecules, used to study and visualize the concentration, distribution and transport of Zn(II) ions in real time. Such agents are designed for high selectivity for zinc in respect to other biological metal ions. However, the issue of their sensitivity to physiological levels of low molecular weight Zn(II) ligands (LMWLs) has not been addressed. We followed the effects of eight

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“…Fluorescein is one of the most widely used fluorophore for biological experiments, since it has a high fluorescence quantum yield of 0.90 or grater in aqueous solution and its excitation and emission wavelengths are in the visible region. Recently, Nagano and coworkers have succeeded in developing fluorescent reagents for the specific detection of nitric oxide (NO), [5][6][7][8] singlet oxygen ( 1 O 2 ), 9,10) and others 11,12) using fluorescein as the fluorophore. 4,4-Difluoro-4-bora-3a,4a-diazas-indacenes (BODIPYs) are also well-known fluorophores that have very sharp and narrow fluorescence bandwidths in addition to their high fluorescence quantum yield in aqueous solution.…”

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confidence: 99%

Improvement of Fluorescence Characteristics of Coumarins: Syntheses and Fluorescence Properties of 6-Methoxycoumarin and Benzocoumarin Derivatives as Novel Fluorophores Emitting in the Longer Wavelength Region and Their Application to Analytical Reagents

Murata

Masuda

Kamochi

et al. 2005

CHEMICAL & PHARMACEUTICAL BULLETIN

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Fluorometric analysis is one of the most sensitive methods for detecting organic and/or inorganic compounds and therefore it has been widely used in many scientific fields with improving analytical instruments such as high-performance liquid chromatography (HPLC), capillary electrophoresis (CE), fluorescence confocal microscopy, etc. For these purposes, a great deal of effort has gone into the development of fluorescent reagents using various fluorophores, which can be utilized for derivatizing biologically active small molecules, labeling macromolecules such as proteins and DNAs, and probing ions such as biologically relevant metal cations and inorganic or organic anions. [1][2][3][4] In the development of such fluorescent reagents, it should first be ensured that the fluorophore used has a high fluorescence quantum yield because of the reliably high sensitivity in the detection. Fluorescein is one of the most widely used fluorophore for biological experiments, since it has a high fluorescence quantum yield of 0.90 or grater in aqueous solution and its excitation and emission wavelengths are in the visible region. Recently, Nagano and coworkers have succeeded in developing fluorescent reagents for the specific detection of nitric oxide (NO), [5][6][7][8] singlet oxygen ( 1 O 2 ), 9,10) and others 11,12) using fluorescein as the fluorophore. 4,4-Difluoro-4-bora-3a,4a-diazas-indacenes (BODIPYs) are also well-known fluorophores that have very sharp and narrow fluorescence bandwidths in addition to their high fluorescence quantum yield in aqueous solution. 4,13,14) Therefore they have been used as mother fluorophores for many analytical purposes. [15][16][17][18] However, the difference between the excitation and emission wavelengths of these derivatives are very small, less than approximately 30 nm for fluoresceins and 15 nm for BODIPYs, and it is necessary to correct their spectra from interferences such as Rayleigh or Raman scattering light.On the other hand, coumarin (2H-benzopyran-2-one), is also another interesting fluorophore, since its fluorescence changes drastically with substituents and their introduced positions. [19][20][21] In previous papers, we reported the fluorescence characteristics of methoxycoumarins and discussed the structural features of strongly fluorescing methoxycoumarins from the viewpoint of intramolecular charge transfer (ICT) between push-and pull-substituents in the ground and the excited states.22,23) Based on the above findings, we succeeded in developing novel fluorescent reagents with both high sensitivity and functions such as self-catalytic reactivity. [24][25][26] In the course of those studies, we suspected that most coumarin derivatives showed large Stokes shifts in their fluorescence spectra as compared with other fluorophores. These large Stokes shifts appear to be advantageous for escaping autofluorescence from biological molecules and reducing the self-absorption of chromophores. Further, if the emission bands of coumarins could be shifted to longer wavelengths with larg...

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Highly Zinc-Selective Fluorescent Sensor Molecules Suitable for Biological Applications

Cited by 337 publications

References 27 publications

Imaging dynamic insulin release using a fluorescent zinc indicator for monitoring induced exocytotic release (ZIMIR)

Imaging dynamic insulin release using a fluorescent zinc indicator for monitoring induced exocytotic release (ZIMIR)

Ternary complex formation and competition quench fluorescence of ZnAF family zinc sensors

Improvement of Fluorescence Characteristics of Coumarins: Syntheses and Fluorescence Properties of 6-Methoxycoumarin and Benzocoumarin Derivatives as Novel Fluorophores Emitting in the Longer Wavelength Region and Their Application to Analytical Reagents

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