Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging
Background: Norbormide (NRB) is a selective rat toxicant endowed with vasoconstrictor activity confined to the rat peripheral arteries. In a recent work we used a fluorescent derivative of NRB (NRB-AF12), obtained by coupling the NBD fluorophore to the parent molecule via a linker, in order to gain...
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Frontiers Media S.A.
2018-09-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphar.2018.01055/full |
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| author | Claudio D'Amore Genny Orso Alessia Forgiarini Giulio Ceolotto David Rennison Giovanni Ribaudo Morgan Jay-Smith Brian Hopkins Margaret A. Brimble Sergio Bova |
| author_facet | Claudio D'Amore Genny Orso Alessia Forgiarini Giulio Ceolotto David Rennison Giovanni Ribaudo Morgan Jay-Smith Brian Hopkins Margaret A. Brimble Sergio Bova |
| author_sort | Claudio D'Amore |
| collection | DOAJ |
| description | Background: Norbormide (NRB) is a selective rat toxicant endowed with vasoconstrictor activity confined to the rat peripheral arteries. In a recent work we used a fluorescent derivative of NRB (NRB-AF12), obtained by coupling the NBD fluorophore to the parent molecule via a linker, in order to gain information about the possible site of action of the unlabeled compound. We found that NRB-AF12 labeled intracellular organelles in both NRB-sensitive and -insensitive cells and we accordingly proposed its use as a scaffold for the development of a new class of fluorescent probes. In this study, we examined the fluorescent properties of a BODIPY FL-conjugated NRB probe (MC009) developed: (A) to verify if NRB distribution could be influenced by the attached fluorophore; (B) to improve the fluorescent performance of NRB-AF12.Methods: MC009 characteristics were investigated by confocal fluorescence microscopy, in freshly isolated rat caudal artery myocytes (FIRCAM) and in LX2 cells, representative of NRB-sensitive and insensitive cells, respectively.Main results: In both FIRCAM and LX2 cells MC009 stained endoplasmic reticulum, mitochondria, Golgi apparatus and lipid droplets, revealing the same intracellular distribution as NRB-AF12, and, at the same time, had both improved photostability and gave a more intense fluorescent signal at lower concentrations than was possible with NRB-AF12, which resulted in a better and finer visualization of intracellular structures. Furthermore, MC009 was effective in cellular labeling in both living and fixed cells. At the concentration used to stain the cells, MC009 did not show any cytotoxic effect and did not affect the regular progression of cell cycle and division.Conclusions: This study demonstrates that the distribution of fluorescently labeled NRB is not affected by the type of fluorophore attached to the parent compound, supporting the idea that the localization of the fluorescent derivatives may reasonably reflect that of the parent compound. In addition, we observed a marked improvement in the fluorescent properties of BODIPY FL-conjugated NRB (MC009) over its NBD-derived counterpart (NRB-AF12), confirming NRB as a scaffold for the development of new, high performance, non-toxic fluorescent probes for the labeling of intracellular structures in both living and fixed cells. |
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| id | doaj.art-ad49c50446db4668944dd62b2acd7d8e |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-20T19:43:41Z |
| publishDate | 2018-09-01 |
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| series | Frontiers in Pharmacology |
| spelling | doaj.art-ad49c50446db4668944dd62b2acd7d8e2022-12-21T19:28:27ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122018-09-01910.3389/fphar.2018.01055409972Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell ImagingClaudio D'Amore0Genny Orso1Alessia Forgiarini2Giulio Ceolotto3David Rennison4Giovanni Ribaudo5Morgan Jay-Smith6Brian Hopkins7Margaret A. Brimble8Sergio Bova9Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, ItalyDepartment of Medicine, University of Padova, Padova, ItalySchool of Chemical Sciences, University of Auckland, Auckland, New ZealandDepartment of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, ItalySchool of Chemical Sciences, University of Auckland, Auckland, New ZealandLandcare Research, Lincoln, New ZealandSchool of Chemical Sciences, University of Auckland, Auckland, New ZealandDepartment of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, ItalyBackground: Norbormide (NRB) is a selective rat toxicant endowed with vasoconstrictor activity confined to the rat peripheral arteries. In a recent work we used a fluorescent derivative of NRB (NRB-AF12), obtained by coupling the NBD fluorophore to the parent molecule via a linker, in order to gain information about the possible site of action of the unlabeled compound. We found that NRB-AF12 labeled intracellular organelles in both NRB-sensitive and -insensitive cells and we accordingly proposed its use as a scaffold for the development of a new class of fluorescent probes. In this study, we examined the fluorescent properties of a BODIPY FL-conjugated NRB probe (MC009) developed: (A) to verify if NRB distribution could be influenced by the attached fluorophore; (B) to improve the fluorescent performance of NRB-AF12.Methods: MC009 characteristics were investigated by confocal fluorescence microscopy, in freshly isolated rat caudal artery myocytes (FIRCAM) and in LX2 cells, representative of NRB-sensitive and insensitive cells, respectively.Main results: In both FIRCAM and LX2 cells MC009 stained endoplasmic reticulum, mitochondria, Golgi apparatus and lipid droplets, revealing the same intracellular distribution as NRB-AF12, and, at the same time, had both improved photostability and gave a more intense fluorescent signal at lower concentrations than was possible with NRB-AF12, which resulted in a better and finer visualization of intracellular structures. Furthermore, MC009 was effective in cellular labeling in both living and fixed cells. At the concentration used to stain the cells, MC009 did not show any cytotoxic effect and did not affect the regular progression of cell cycle and division.Conclusions: This study demonstrates that the distribution of fluorescently labeled NRB is not affected by the type of fluorophore attached to the parent compound, supporting the idea that the localization of the fluorescent derivatives may reasonably reflect that of the parent compound. In addition, we observed a marked improvement in the fluorescent properties of BODIPY FL-conjugated NRB (MC009) over its NBD-derived counterpart (NRB-AF12), confirming NRB as a scaffold for the development of new, high performance, non-toxic fluorescent probes for the labeling of intracellular structures in both living and fixed cells.https://www.frontiersin.org/article/10.3389/fphar.2018.01055/fullfluorescent probeslive cell imagingnorbormideBODIPYvascular smooth muscle cellsLX2 cells |
| spellingShingle | Claudio D'Amore Genny Orso Alessia Forgiarini Giulio Ceolotto David Rennison Giovanni Ribaudo Morgan Jay-Smith Brian Hopkins Margaret A. Brimble Sergio Bova Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging Frontiers in Pharmacology fluorescent probes live cell imaging norbormide BODIPY vascular smooth muscle cells LX2 cells |
| title | Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging |
| title_full | Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging |
| title_fullStr | Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging |
| title_full_unstemmed | Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging |
| title_short | Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging |
| title_sort | synthesis and biological characterization of a new norbormide derived bodipy fl conjugated fluorescent probe for cell imaging |
| topic | fluorescent probes live cell imaging norbormide BODIPY vascular smooth muscle cells LX2 cells |
| url | https://www.frontiersin.org/article/10.3389/fphar.2018.01055/full |
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