New properties of a bioinspired pyridine benzimidazole compound as a novel differential staining agent for endoplasmic reticulum and Golgi apparatus in fluorescence live cell imaging

Felipe M. Llancalahuen, Juan A. Fuentes, Alexander Carreño, César Zúñiga, Dayán Páez-Hernández, Manuel Gacitúa, Rubén Polanco, Marcelo D. Preite, Ramiro Arratia-Pérez, Carolina Otero

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Abstract

In this study, we explored new properties of the bioinspired pyridine benzimidazole compound B2 (2,4-di-tert-butyl-6-(3H-imidazo[4,5-c]pyridine-2-yl)phenol) regarding its potential use as a differential biomarker. For that, we performed 1D 1HNMR (TOCSY), UV-Vis absorption spectra in different organic solvents, voltammetry profile (including a scan-rate study), and TD-DFT calculations that including NBO analyses, to provide valuable information about B2 structure and luminescence. In our study, we found that the B2 structure is highly stable, where the presence of an intramolecular hydrogen bond (IHB) seems to have a crucial role in the stability of luminescence, and its emission can be assigned as fluorescence. In fact, we found that the relatively large Stokes Shift observed for B2 (around 175 nm) may be attributed to the stability of the B2 geometry and the strength of its IHB. On the other hand, we determined that B2 is biocompatible by cytotoxicity experiments in HeLa cells, an epithelial cell line. Furthermore, in cellular assays we found that B2 could be internalized by passive diffusion in absence of artificial permeabilization at short incubation times (15 min to 30 min). Fluorescence microscopy studies confirmed that B2 accumulates in the endoplasmic reticulum (ER) and Golgi apparatus, two organelles involved in the secretory pathway. Finally, we determined that B2 exhibited no noticeable blinking or bleaching after 1 h of continuous exposure. Thus, B2 provides a biocompatible, rapid, simple, and efficient way to fluorescently label particular organelles, producing similar results to that obtained with other well-established but more complex methods.

Original languageEnglish
Article number345
JournalFrontiers in Chemistry
Volume6
Issue numberAUG
DOIs
Publication statusPublished - 15 Aug 2018

Fingerprint

Luminescence
Hydrogen bonds
Fluorescence
Imaging techniques
Fluorescence microscopy
Biomarkers
Voltammetry
Cytotoxicity
Bleaching
Phenol
Discrete Fourier transforms
Organic solvents
Labels
Absorption spectra
Assays
Geometry
Experiments
benzimidazole
pyridine
Epithelial Cells

Keywords

  • Benzimidazole
  • Differential staining
  • Endoplasmic reticulum
  • Fluorescence
  • Golgi apparatus
  • Hydrogen bond

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

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title = "New properties of a bioinspired pyridine benzimidazole compound as a novel differential staining agent for endoplasmic reticulum and Golgi apparatus in fluorescence live cell imaging",
abstract = "In this study, we explored new properties of the bioinspired pyridine benzimidazole compound B2 (2,4-di-tert-butyl-6-(3H-imidazo[4,5-c]pyridine-2-yl)phenol) regarding its potential use as a differential biomarker. For that, we performed 1D 1HNMR (TOCSY), UV-Vis absorption spectra in different organic solvents, voltammetry profile (including a scan-rate study), and TD-DFT calculations that including NBO analyses, to provide valuable information about B2 structure and luminescence. In our study, we found that the B2 structure is highly stable, where the presence of an intramolecular hydrogen bond (IHB) seems to have a crucial role in the stability of luminescence, and its emission can be assigned as fluorescence. In fact, we found that the relatively large Stokes Shift observed for B2 (around 175 nm) may be attributed to the stability of the B2 geometry and the strength of its IHB. On the other hand, we determined that B2 is biocompatible by cytotoxicity experiments in HeLa cells, an epithelial cell line. Furthermore, in cellular assays we found that B2 could be internalized by passive diffusion in absence of artificial permeabilization at short incubation times (15 min to 30 min). Fluorescence microscopy studies confirmed that B2 accumulates in the endoplasmic reticulum (ER) and Golgi apparatus, two organelles involved in the secretory pathway. Finally, we determined that B2 exhibited no noticeable blinking or bleaching after 1 h of continuous exposure. Thus, B2 provides a biocompatible, rapid, simple, and efficient way to fluorescently label particular organelles, producing similar results to that obtained with other well-established but more complex methods.",
keywords = "Benzimidazole, Differential staining, Endoplasmic reticulum, Fluorescence, Golgi apparatus, Hydrogen bond",
author = "Llancalahuen, {Felipe M.} and Fuentes, {Juan A.} and Alexander Carre{\~n}o and C{\'e}sar Z{\'u}{\~n}iga and Day{\'a}n P{\'a}ez-Hern{\'a}ndez and Manuel Gacit{\'u}a and Rub{\'e}n Polanco and Preite, {Marcelo D.} and Ramiro Arratia-P{\'e}rez and Carolina Otero",
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T1 - New properties of a bioinspired pyridine benzimidazole compound as a novel differential staining agent for endoplasmic reticulum and Golgi apparatus in fluorescence live cell imaging

AU - Llancalahuen, Felipe M.

AU - Fuentes, Juan A.

AU - Carreño, Alexander

AU - Zúñiga, César

AU - Páez-Hernández, Dayán

AU - Gacitúa, Manuel

AU - Polanco, Rubén

AU - Preite, Marcelo D.

AU - Arratia-Pérez, Ramiro

AU - Otero, Carolina

PY - 2018/8/15

Y1 - 2018/8/15

N2 - In this study, we explored new properties of the bioinspired pyridine benzimidazole compound B2 (2,4-di-tert-butyl-6-(3H-imidazo[4,5-c]pyridine-2-yl)phenol) regarding its potential use as a differential biomarker. For that, we performed 1D 1HNMR (TOCSY), UV-Vis absorption spectra in different organic solvents, voltammetry profile (including a scan-rate study), and TD-DFT calculations that including NBO analyses, to provide valuable information about B2 structure and luminescence. In our study, we found that the B2 structure is highly stable, where the presence of an intramolecular hydrogen bond (IHB) seems to have a crucial role in the stability of luminescence, and its emission can be assigned as fluorescence. In fact, we found that the relatively large Stokes Shift observed for B2 (around 175 nm) may be attributed to the stability of the B2 geometry and the strength of its IHB. On the other hand, we determined that B2 is biocompatible by cytotoxicity experiments in HeLa cells, an epithelial cell line. Furthermore, in cellular assays we found that B2 could be internalized by passive diffusion in absence of artificial permeabilization at short incubation times (15 min to 30 min). Fluorescence microscopy studies confirmed that B2 accumulates in the endoplasmic reticulum (ER) and Golgi apparatus, two organelles involved in the secretory pathway. Finally, we determined that B2 exhibited no noticeable blinking or bleaching after 1 h of continuous exposure. Thus, B2 provides a biocompatible, rapid, simple, and efficient way to fluorescently label particular organelles, producing similar results to that obtained with other well-established but more complex methods.

AB - In this study, we explored new properties of the bioinspired pyridine benzimidazole compound B2 (2,4-di-tert-butyl-6-(3H-imidazo[4,5-c]pyridine-2-yl)phenol) regarding its potential use as a differential biomarker. For that, we performed 1D 1HNMR (TOCSY), UV-Vis absorption spectra in different organic solvents, voltammetry profile (including a scan-rate study), and TD-DFT calculations that including NBO analyses, to provide valuable information about B2 structure and luminescence. In our study, we found that the B2 structure is highly stable, where the presence of an intramolecular hydrogen bond (IHB) seems to have a crucial role in the stability of luminescence, and its emission can be assigned as fluorescence. In fact, we found that the relatively large Stokes Shift observed for B2 (around 175 nm) may be attributed to the stability of the B2 geometry and the strength of its IHB. On the other hand, we determined that B2 is biocompatible by cytotoxicity experiments in HeLa cells, an epithelial cell line. Furthermore, in cellular assays we found that B2 could be internalized by passive diffusion in absence of artificial permeabilization at short incubation times (15 min to 30 min). Fluorescence microscopy studies confirmed that B2 accumulates in the endoplasmic reticulum (ER) and Golgi apparatus, two organelles involved in the secretory pathway. Finally, we determined that B2 exhibited no noticeable blinking or bleaching after 1 h of continuous exposure. Thus, B2 provides a biocompatible, rapid, simple, and efficient way to fluorescently label particular organelles, producing similar results to that obtained with other well-established but more complex methods.

KW - Benzimidazole

KW - Differential staining

KW - Endoplasmic reticulum

KW - Fluorescence

KW - Golgi apparatus

KW - Hydrogen bond

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U2 - 10.3389/fchem.2018.00345

DO - 10.3389/fchem.2018.00345

M3 - Article

VL - 6

JO - Frontiers in Chemistry

JF - Frontiers in Chemistry

SN - 2296-2646

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