Multi-coloured fluorescent sensing toolbox for selective detection of nitroxyl in vitro and ex vivo

V. Staikopoulos; X. Zhang; B. P. Pullen; P. Reineck; A. K. Vidanapathirana; S. M. Lee; J. Liu; C. Bursill; M. R. Hutchinson; A. D. Abell

doi:10.1039/D2SD00006G

Multi-coloured fluorescent sensing toolbox for selective detection of nitroxyl in vitro and ex vivo†

V. Staikopoulos,‡^ab X. Zhang,‡^bc B. P. Pullen,^abd P. Reineck,

^be A. K. Vidanapathirana,^abd S. M. Lee,^a J. Liu,^a C. Bursill,^abd M. R. Hutchinson^ab and A. D. Abell

*^bc

Author affiliations

* Corresponding authors

^a Adelaide Medical School, University of Adelaide, Adelaide, South Australia 5005, Australia

^b Australian Research Council (ARC) Centre of Excellence for Nanoscale BioPhotonics, Adelaide Medical School, University of Adelaide, Adelaide, South Australia, Australia

^c Department of Chemistry, University of Adelaide, Adelaide, South Australia, Australia
E-mail: andrew.abell@adelaide.edu.au

^d Vascular Research Centre, Lifelong Health Theme, South Australian Health and Medical Research Institute (SAHMRI), Adelaide, South Australia 5000, Australia

^e School of Science, RMIT University, Melbourne, Victoria, 3001 Australia

Abstract

Methods for the endogenous detection of nitroxyl (azanone; HNO), the reduced and protonated derivative of nitric oxide (NO), are required to define its cardiovascular function and its key role in chronic pain. This study reports the design, synthesis and biological evaluation of 3 super-bright, highly sensitive, specific and non-cytotoxic arylphosphine-based fluorescent nitroxyl sensors that enable the detection of endogenous nitroxyl in vitro and ex vivo. The presence of endogenous nitroxyl is observed in the murine microglial cell line (BV2), primary human coronary artery endothelial cells (HCAECs) and rat cardiomyocyte cell line (H9C2) by spectroscopy and/or confocal microscopy following the addition of relevant stimulants. Furthermore, the presence of endogenous nitroxyl is detected in rat blood samples. The novel sensor 3 (rhodol 4 on a 2-(diphenylphosphanyl)benzoate backbone) is the most sensitive in detecting endogenous nitroxyl following stimulation, in the presence of biological media such as HBSS and DMEM. This then provides a validated tool for detecting nitroxyl in biological systems.

Sensors & Diagnostics

Multi-coloured fluorescent sensing toolbox for selective detection of nitroxyl in vitro and ex vivo†

Abstract

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Multi-coloured fluorescent sensing toolbox for selective detection of nitroxyl in vitro and ex vivo

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