Intracellular viscosity is an essential microenvironmental parameter and HS is a critical gaseous signaling molecule, which are both related to various physiological processes. It is reported that the change of viscosity and an imbalance of HS production in the mitochondria are both associated with overexpression of amyloid betapeptide (Aβ), which is thought to play a central role in the pathogenesis of Alzheimer's disease (AD). However, to our best knowledge, no fluorescent probe is found for dual detection of mitochondrial viscosity and HS. Herein, a dual-response fluorescent probe (Mito-VS) is designed and synthesized to monitor the level of viscosity and HS, respectively. Mito-VS itself is nonfluorescent due to a free intramolecular rotation between dimethylaniline and pyridine. After the increase of viscosity, the rotation is prohibited and an intense red fluorescence is released. Upon the addition of HS, the probe can react with HS to form compound 3 and a strong green fluorescence can be observed. Moreover, the probe possesses a good mitochondrion-targeting ability and is applied for imaging the change of viscosity on the red channel and visualizing the variation of exogenous and endogenous HS concentration on the green channel in mitochondria. Most importantly, the probe is capable of studying the cross-talk influence of viscosity and HS in mitochondria, which is very beneficial for knowing the pathogenesis of AD.
Alkaline phosphatase (ALP) is an essential enzyme and widely distributes in a variety of tissues. To date, various nanomaterial and small-molecule fluorescent probes for ALP have been constructed successfully, but the emission wavelengths of these probes are in the ultraviolet or visible range, which is not beneficial for bioimaging. Herein, a hemicyanine-based near-infrared (NIR) fluorescent probe named CyP is first synthesized and used to detect ALP activity. The characteristics of probe CyP are as follows: (1) The probe possesses a facile structure, which can be obtained by easy synthetic steps. (2) The fluorescence emission of the sensing system is at 738 nm belonging to NIR region, which is suitable for bioimaging in vivo. (3) The probe exhibits high sensitivity to ALP with 10-fold fluorescence enhancement and low detection limit (0.003 U/mL) can match the level of ALP in vivo. (4) The fluorescent change of the probe is attributed to the fact that ALP-catalyzed cleavage of the phosphate group in CyP induces the transformation of CyP (fluorescence off) into CyOH (fluorescence on), which is proved by HPLC, P NMR, MS, and DFT calculation. (5) The NIR fluorescent probe is applied for the detection of endogenous ALP activity in various biological samples such as cell, tissue, and living animal with satisfactory results.
Nitric
oxide (NO) is a vital gaseous signal molecule and plays
an important role in diverse physiological and pathological processes
including regulation of vascular functions. Endoplasmic reticulum
(ER) stress is caused by the accumulation of misfolded or unfolded
protein in the ER. Besides, ER stress induced by NO can be involved
in the pathogenesis of various vascular diseases. Unfortunately, to
the best of our knowledge, no ER-targeting probe for NO is reported
to study the relationship between ER stress and the level of NO in
a biological system. Herein, an ER-targeted fluorescent probe named
ER-Nap-NO for imaging of NO is designed and synthesized. ER-Nap-NO
consists of three main parts: naphthalimide (two-photon fluorophore), o-phenylenediamino (NO recognition group), and methyl sulfonamide
(ER-targetable group). The probe itself is nonfluorescent because
a photoinduced electron transfer (PET) process exists. After the addition
of NO, the PET process is inhibited and thus strong fluorescence is
released. Moreover, the response mechanism is confirmed by 1H NMR and mass spectra and DFT calculation in detail. In addition,
from the experimental results, we can conclude that the probe displays
several obvious advantages including high sensitivity, selectivity,
and ER-targetable ability. Based on these excellent properties, the
probe is used for the two-photon imaging of exogenous and endogenous
NO in ER of living cells. Most importantly, the ER-targetable probe
has potential capability as a tool for investigating the level of
NO during tunicamycin-induced ER stress in cells and tissues, which
is beneficial for revealing the role of NO in ER-associated vascular
diseases.
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