An Activatable and Switchable Nanoaggregate Probe for Detecting H2S and Its Application in Mice Brains

Article abstract of DOI:10.1002/asia.202000964

Employing a sequentially activated probe design method, an activatable, switchable and dual-mode probe was designed. This nanoprobe, HSDPP, could be effectively activated by H₂S to form H-type aggregates with green emission; subsequently, the aggregates could bind to mtDNA to form monomers and the emIssion color switched from green to deep-red. We exploited HSDPP to image exogenous and endogenous H₂S in living cells. Of note, for the first time, this novel nanoprobe with an optimal partition coefficient value (LogP=1.269) was successfully applied for tracking the endogenous H₂S upregulation stimulated by cystathionase activator S-adenosyl-L-methionine (SAM) in mice brains. Finally, our work provides an invaluable chemical tool for probing endogenous H₂S upregulation in vitro/vivo and, importantly, affords a prospective design strategy for developing switchable chemosensors to unveil the relationship between biomolecules and DNA in mitochondria in many promising areas.

Full text of DOI:10.1002/asia.202000964

CHEMISTRY
AN ASIAN JOURNAL
www.chemasianj.org
Accepted Article
Title: A novel activatable and switchable nanoaggregate probe for
detecting H2S and its application in mice brains
Authors: Ya-Lin Qi, Li-Li Chen, Long Guo, Yu-Shun Yang, Zhen-Xiang
He, and Hai-Liang Zhu
This manuscript has been accepted after peer review and appears as an
Accepted Article online prior to editing, proofing, and formal publication
of the final Version of Record (VoR). This work is currently citable by
using the Digital Object Identifier (DOI) given below. The VoR will be
published online in Early View as soon as possible and may be different
to this Accepted Article as a result of editing. Readers should obtain
the VoR from the journal website shown below when it is published
to ensure accuracy of information. The authors are responsible for the
content of this Accepted Article.
To be cited as: Chem. Asian J. 10.1002/asia.202000964
Link to VoR: https://doi.org/10.1002/asia.202000964
A Journal of
A sister journal of Angewandte Chemie
and Chemistry – A European Journal

10.1002/asia.202000964
Chemistry - An Asian Journal
A novel activatable and switchable nanoaggregate probe for detecting
H2S and its application in mice brains
Ya-Lin Qi, # Li-Li Chen, # Long Guo, Chen-Wen Shao, Ya-Ni Liu, Yu-Shun Yang,* Zhen-Xiang He,*
and Hai-Liang Zhu*
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University,
Nanjing, 210023, China.
E-mail: zhuhal@nju.edu.cn; ys_yang@nju.edu.cn; zxhe@nju.edu.cn.
# Both authors contributed equally to this work.
Abstract: Employing a sequentially activated
probe design method, an activatable, switchable
and dual-mode probe was designed. This
nanoprobe, HSDPP, could be effectively
activated by H2S to form H-type aggregates with
green emission; subsequently, the aggregates
could bind to mtDNA to form monomers and the
emssion color switched from green to deep-red.
We exploited HSDPP to image the exogenous
and endogenous H2S in living cells. Of note, for
the first time, this novel nanoprobe with an
optimal partition coefficient value (LogP =
1.269) was successfully applying for tracking the
endogenous H2S upregulation stimulated by
cystathionase activator S-adenosyl-L-methionine
(SAM) in mice brains. Finally, our work
provides an invaluable chemical tool for probing
endogenous H2S upregulation in vitro/vivo and,
importantly, affords a prospective design
strategy for developing switchable chemosensors
to unveil the relationship between the
biomolecules and DNAin mitochondrial in many
promising areas.
Introduction
Molecular probes, which have been widely
utilized to track biomarkers and molecular events
in living biosystems, can be broadly categorized
into two fields including “always on” tags and
activatable sensors.[1] “Always on” tags produce
contrast signals through accumulation and,
particularly, their signals keep stable within the
position or situation of interest.[2] On the contrary,
serving as molecular beacons or optical switches,
activatable probes are commonly in the “off”
state at the beginning and can only be stimulated
by the certain target to send out corresponding
signals.[3,4]
In
comparison
with
the
nonresponsive species ("always off") and the
"always on" tags, the activatable probes benefit
from the signal changes, which further lead to the
1
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
advantages including higher signal-to-noise ratio,
lower detection of limit, permitting sensitive and
so on.[5–7] Therefore, they are capable of
responding specifically to the biomarkers of
diseases and allow disease-specific diagnosis.[8]
Accordingly, investigators have paid an
increasing attention to the activatable probes in
recent years.[9–12]
based signal transmission,[27] and so on. One step
further, we still hope that the signal switching
could be regulated by an "all-or-nothing"
biomolecule which itself is the marker of specific
physiological or pathological models, for
[28]
example, glutathione in solid tumor
or
caspase-3 in apoptosis.[29] For the monitoring of
H2S, the requirements become specialized
because the internal H2S is usually at the
micromole level.[30] Thus, in spite of the high
sensitivity, the major concern is introducing
stable NIR signals under a model-specific
regulation. Accordingly, we put our attention on
On the other hand, as the third multifunctional
gasotransmitter, H2S has been recognized as an
important
signaling
molecule
in
the
cardiovascular and nervous systems.[13,14] There
is not much to debate anymore about the
significance on monitoring the internal level of
H2S,[15–18] however, when the investigations
nowadays come to the tissues or models in vivo,
near-infrared (NIR) implements seem more
favoured due to the strong penetrability and
photothermal/photoacoustic effect.[19] No matter
the probes are small molecular or nanoscale
ones,[20,21] one major concern is maintaining
obvious reporting signals to guarantee both
sensitivity and selectivity. Basically, longer
wavelength means weaker energy, thus a direct
NIR sensing usually need a subsequent
procedure of amplification. Common enlarging
strategies include instrumental calibration,[22]
Aggregation-Induced Emission (AIE),[23,24]
protein-assistant amplification,[25,26] nanocarrier-
Mitochondrial DNA (mtDNA).[31] As
a
significant biomarker in bioenergetics and
biochemistry, mtDNA shows at least two
favourable features for acting as the signal
switcher. One is that mtDNA is ubiquitously
existing in eukaryotic cells with a certain level.[32]
The other is that mtDNA is associated with
several
inherited
diseases,
including
inflammation, autoimmunity, and aging.[33–36]
Among these diseases, the mtDNA level is
remarkably affected in specific positions such as
brain and heart, while these positions are also the
key ones for H2S-associated diagnose.[37–39]
Although
a
variety of small molecular
fluorescent probes have been reported for
detecting H2S or mtDNA independently,[40–43] in
2
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
this work, we attempted to combine them into a
detecting system which was activatable,
switchable and available for applications in
brains.
Herein, we exploited a novel activatable and
switchable probe, HSDPP, for monitoring the
H2S level in vitro and in vivo with two different
channels (Scheme 1). Consulting the recognition
Scheme 1. The design strategy and sensing mode of HSDPP in this work. a) The previous work of probe HS-CyBz
for detecting H2S; b) The previous work of probe MNQI for interacting with DNA. c) The structure and sensing
mode of HSDPP.
group from HS-CyBz (which seemed more
selective with smaller steric hindrance) and the
binding group from MNQI (which interacted
with mtDNA), HSDPP was designed to achieve
the step-by-step detecting system. When initially
activated by H2S, HSDPP presented strong
green fluorescence via the generation of HSDP
and H-type aggregates. Subsequently, the
interaction between HSDP and mtDNA resulted
in the confirmation of monomers, which
switched the fluorescent signal into the red
channel with a practical intensity. Therefore, this
strategy could efficiently avoid the interference
from spectral overlap, and effectively trace
endogenous H2S upregulation promoted by
cystathionase activator SAM in mice brains for
3
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
the first time. More than introducing a novel
activatable implement, this work brought a
mtDNA-regulated switching strategy for
transferring fluorescent signals into NIR region,
which opened a new window for developing
model-specific reporters as well as revealing the
crosstalk between target molecules and mtDNA
in physiological and pathological prodecures.
Results and Discussion
NMR, 13C NMR and mass spectrometry (MS) of
HSDPP, the activated probe HSDP and
intermediates were characterized in Figures S1-
S12.
The proposed sensing mechanism of HSDPP
for sequentially visualizing H2S and mtDNAwas
shown in Scheme 1c. We hypothesized that the
free probe initially exhibited weak fluorescence
due to the prevention from the excited-state
intramolecular proton-transfer (ESIPT) process
by ester. However, the nucleophilic substitution
on ester by H2S would release the hydroxyl
group of HBT to induce ESIPT, lead to the
formation of the H-type aggregates (the activated
probe HSDP) and lighted up the platform,
accompanied by a shorter emission wavelength;
subsequently, HSDP would bind to mtDNA to
Design and Recognition Mechanism Studies.
In HSDPP, iodobenzoic acid methyl ester was
selected as the reaction site due to its high
selectivity and sensitivity towards H2S,[40] as
shown in Scheme 1a. For mtDNA, the cationic
quinolinium group was served as the part of
binding unit,[44] which could be also anchored in
the mitochondrial (Scheme 1b). In addition, the
quinolinium unit could contribute to the
aggregation-induced-emission (AIE) feature of
the nanoaggregates to some extent.[8] As for the
form monomers, which resulted in
a
bathochromic shift with far red fluorescence
emission. Therefore, this nanoprobe could
realize the detection of H2S regulated by mtDNA
in biological applications, with multi-channel
fluorescent signals.
fluorophore,
2-(2-hydroxy-phenyl)
benzothiazole (HBT), a universal platform,
served as the scaffold ascribed to its unique
properties (Scheme 1c). It included advantages
such as large Stokes shift, environmental
sensitivity and potential for ratiometric
sensing.[45] The molecular probe HSDPP was
synthesized according to the route shown in
Spectroscopic Analysis of Probe HSDPP
towards H2S and Mitochondrial DNA. With
the probe HSDPP on hand, the spectral
properties of HSDPP in the absence or presence
of H2S (Na2S as the donor) and mtDNA (calf
thymus DNA (ctDNA) as a model) were
1
Supporting Information (Scheme S1). The H
4
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
determined. As expected, the inactivated probe
HSDPP (10 μM) alone presented very weak
fluorescence and absorbance (Figure 1a and 1b).
However, the addition of 200 μM H2S to the
solution of HSDPP produced a strong green
fluorescence with a peak at 550 nm when excited
at 443 nm (Figure 1d), and the equilibrium was
reached within 55 min with a 35-fold
fluorescence enhancement (Figures 1a, 1c, 1d
and S13). At the same time, a remarkable
absorbance band at 628 nm was observed (Figure
0 to 220 μM H2S treatment (Figures 1c and S13),
and the detection limit was calculated to as 0.124
μM (Figures S14). At this stage, no deep-red
signal was obtained (Figure 1a). Further, upon
the addition of ctDNAto the above solution, only
a slight green fluorescence enhancement could
be observed, while the absorption spectrum
displayed negligible changes (Figures 1a and 1b).
This results suggested that ctDNA itself could
not trigger on HSDPP fluorescence and the
green emission enhancement was caused by H2S
alone.
1b). Moreover,
a
concentration-dependent
fluorescent enhancement could be obtained from
Figure. 1 Response of HSDPP towards H2S and HSDP towards ctDNA, respectively. a) Fluorescence emission
spectra of HSDPP (10 µM) in the presence or presence of ctDNA (3500 μM)/H2S (200 μM); b) The absorption
spectra of HSDPP (10 µM) in the absence or presence of ctDNA (3500 μM)/H2S (200 μM); c) Fluorescence emission
spectra of HSDPP after treatment with H2S for different incubation concentration (0–220 μM). Inset: Time-dependent
changes in the fluorescence intensity of HSDPP at 550 nm; d) Fluorescence excitation spectra of HSDPP (10 µM)
in the presence or presence of H2S (200 μM) and HSDP in the presence or presence of ctDNA (3500 μM); e)
5
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
Fluorescence emission spectra of HSDP after treatment with ctDNA for different incubation concentration (0–4000
μM); f) The absorption spectra of HSDP in the absence or presence of ctDNA (3500 μM). All data were acquired in
phosphate-buffered saline (PBS) buffer (10 mM, pH 7.4, 1% DMSO) at 37 °C with excitation at 443 nm, the data
represent the average of three independent experiments.
However, addition of ctDNA (3.5 M) to the
solution of activated probe HSDP (10 μM) to
form monomers yielded more than 10-fold
fluorescence enhancement after being excited at
443 nm (Figure 1d), and the absorbance band at
530 nm was dramatically generated (Figures 1e,
S15 and 1f). These results confirmed that
HSDPP only banded to ctDNAafter activated by
H2S.
for visualizing H2S through the regulating by
mtDNA in live cells and in vivo with different
imaging channels.
To confirm the sensing mechanism, the
products of HSDPP reacted with H2S were
further confirmed by Electrospray ionization
mass spectrometry (ESI-MS) and Ultra
performance liquid chromatography coupled
with mass spectrometry (UPLC/Triple TOF-
MS/MS) (Figure S17 and S18).
Isothermal titration calorimetry (ITC) assay
showed that the binding constant KD was
3.03×10-5 mol-1 and the mtDNA/HSDP
stoichiometry was 2:1 (Figure S16). In addition,
the quantum yield (φ) of HSDP and HSDPP
were 0.0372 (reference: methylene blue (MB))
and 0.00197 (reference: rhodamine B),
separately. Furthermore, the detecting system
had a large signal resolution of 100 nm between
the fluorescence peaks of H2S and ctDNA
responses, which reduced the mutual
interference between the two fluorescent peaks
and the occurrence of cross-color during cell
imaging. Thus, based on this desirable results,
the nanoprobe HSDPP indicated great potential
AIE feature of activated probe HSDP. To
confirm the AIE feature of the activated probe
HSDP, we initially conducted the spectral
experiment of HSDP in organic solvent (i.e.,
dimethyl sulfoxide (DMSO)) with different
water content. For the absorption spectra, with
the increase of water content, the blue-shifted
absorption peak at about 650 nm gradually
decreased (Figure S19a). This phenomenon
reflected a typical formation of H-type
aggregation.[46,47] As for the fluorescence spectra,
a
certain aggregation-induced quenching
phenomenon was also observed, which further
indicated that the increase of water content
6
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
would cause the aggregation (Figure S19b). Next,
in order to ensure whether the aggregation of
HSDP formed nanoparticles in aqueous solution,
dynamic light scattering (DLS) and transmission
electron microscope (TEM) experiments were
performed. Evidently, the sizes of the aggregates
formed by HSDP were confirmed to be about
140−190 nm by TEM, while their spherical
morphology was also apparently observed
(Figure 2a). Besides, DLS results clearly showed
that the aggregates displayed a uniform particle
size distribution in Figure 2b. However, no
aggregates about 140-190 nm was found when
ctDNA (3500 μM) was added to the solution of
HSDP by TEM and DLS (Figure S20). Those
results indicated that the aggregates could bond
to mtDNA to form monomers in solutions.
Selectivity and Stability of probe HSDPP. To
investigate the selectivity and anti-interference
ability of HSDPP towards H2S, the fluorescence
changes of HSDPP were also observed in the
presence of various relevant species (Figures. 3a
and 3b). We found that the strongest response
was only towards H2S, while negligible emission
changes were found for other relevant reactive
thiols and bio-molecules. This results indicating
that the probe was a highly selective sensor for
H2S.
To test the response of this nanoprobe under
physiological conditions, the probe was exposed
in different pH buffers from 3.0 to 12.0.
Accordingly, a relatively wide range of pH
applications was observed (Figure S21).
Therefore, those results implied that HSDPP
could be only activated by H2S in living systems
and was stable in mitochondria which possess a
weakly alkaline environment.
Imaging exogenous and endogenous H2S in
living HeLa cells. Before cell imaging
applications, the colorimetric methyl thiazolyl
tetrazolium (MTT) assay was conducted to study
Figure. 2 a) TEM image of HSDP in aqueous solution.
b) DLS determination for hydrodynamic diameter of
HSDP in aqueous solution. The scale bar is 500 nm.
the cytotoxicity of HSDPP on HeLa cells. As
shown in Figure S22, treatment with 0.5–6.0 μM
HSDPP for 9 h, 12 h and 24 h respectively
brought negligible toxicity to living cells, as the
cell viability was all above 80%. Thus, the
working condition of 5 μM HSDPP in 1 h was
safe for physiological applications. We then
applied this nanoprobe to monitor exogenous and
7
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
endogenous H2S in HeLa cells (Figure 4). It was
reported that exogenous Cys could stimulate
cells to produce endogenous H2S in HeLa
cells.[48] As we expected, the green and red
fluorescence in group c was more obvious,
compared with the control group a and b, which
implied that HSDPP could be used to image
endogenous and basal H2S in HeLa cells. On the
in living cells.[49] Interestingly, the deep-red
signal was distinctly decreased and the green
channel presented a slight decreased (Figures d1-
d4). This could be ascribed to the reduction of the
aggregate-type HSDP resulted from the
inhibition of H2S production by NEM, which
suppressed the transformation of aggregates to
the monomers. Thus, the meaningful changing
process of different HSDP types was visualized
in living cells (Figure f). Finally, group e
indicated that the nanoprobe was also able to
detect exogenous H2S in HeLa cells. As a whole,
these above results suggested that HSDPP was
capable of tracking basal H2S regulated by
mtDNA and monitoring the changes of H2S
levels in live cells. The photostability of HSDPP
was also examined in HeLa cells, suggesting
excellent stability of fluorescence and good
performance for long-term traceability in living
cells (Figure S23).
Figure. 3 Probe HSDPP (10 μM) with different
analytes. a) Probe HSDPP (10 μM) with different single
analytes. b) Probe HSDPP (10 μM) responds to 200 μM
H2S in the presence of various analytes, respectively. (1)
-
-
-
CH3COO- (2) SCN-, (3) NO3 , (4) NO2 , (5) H2PO4 , (6)
2-
2-
2-
2-
2-
2-
SO4 , (7) SO3 , (8) S2O3 , (9) S2 , (10) S4 , (11) CO3 ,
(12) K+, (13) Ca2+, (14) Na+, (15) Mg2+ , (16) Hcy, (17)
GSH, (18) Cys, (19) Ser, (20) Leu, (21) H2O2, (22) ClO-,
(23) Blank, (24) H2S. Concentration: 1 mM for Hcy,
GSH and Cys. 200 μM for other analytes. All data
represent the fluorescence intensities at 550 nm. Data
represent the mean ± S.D. of three independent
experiments.
Fluorescence Co-localization Experiments. To
investigate the subcellular distribution of
HSDPP, the HeLa cells pretreated with Cys were
co-stained with HSDPP and different
commercial organelle-targeting agents (Figures
5, S24 and S25). As we expected, the
fluorescence of HSDPP in green and red channel
overlapped better with that of Mito-Tracker Blue
in blue channel (Pearson’s coefficients were
contrary, N-ethylmaleimide (NEM) was
incubated with HeLa cells to investigate that the
nanoprobe HSDPP could selectively detect H2S
8
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
Figure. 4 Cell imaging of probe HSDPP (5 μM). a) Control cells. b) Cells incubated with HSDPP for 60 min; c)
Cells incubated with Cys (1 mM) and then HSDPP; d) Cells incubated with H2S (200 μM) and then HSDPP; e) Cells
incubated with NEM (1 mM) and then HSDPP; f) The relative fluorescence intensity of group a-f. All confocal
imaging was taken after 30 min of pre-processing; λex: 458 nm; Green channel: 400-500 nm; Red channel: 500-600
nm. The scale bar is 20 μm.
thus it improved contrast and depth of tissue
0.88449 and 0.0.72067, respectively) than with
that of lysosome and nucleus (Pearson’s
coefficient were 0.66008/0.65479 and -0.17363/-
0.11819). In addition, the spatial distribution of
the blue, green and red channels in HeLa cells also
tended to be consistent (Figure. 5b). The results
suggested that HSDPP had good cell-membrane
permeability, and it was an excellent fluorescent
tool for accurately displaying H2S levels and
anchoring in the mitochondrial apparatus of cells.
Imaging upregulation of H2S in mice brains.
The activated probe HSDP binding to mtDNA
afforded a deep-red fluorescence emission ( λem
= 650 nm) in the near−infrared (NIR) window,
imaging.[50] Importantly, the log P values of
HSDP and HSDPP were 1.270 and 1.269
separately in octanol/water system, which
implied the potential HSDP and HSDPP for
effectively crossing the blood–brain barrier
(BBB) to achieve imaging in the brain region
with the brain-targeted characteristic. Therefore,
we assessed HSDPP as a NIR imaging probe for
tracking the endogenous H2S upregulation
“regulated” by mtDNA in mice brains. The mice
were performed direct intraperitoneal (i. p.)
injection of 0 SAM (200 μL, 100 mg/kg) or
saline (200 μL) for 12 h, followed by intravenous
(i.v.) injection of HSDPP (100 μL, 10 μmol/L).
9
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
The in vivo and ex vivo fluorescence images of
IVIS imaging system (Figures 6 and S26). We
observed an obvious increase of fluorescence in
mice brains were obtained about 1 h later with an
Figure. 5 Fluorescence co-localization experiment. a) Mito-Tracker (MitoLiteTM Blue FX490) and HSDPP; b) Lift:
Intensity profile of linear region of interest across in the HeLa cells co-stained with Mito-Tracker and HSDPP. Right:
Hela cells co-stained with Mito-Tracker and HSDPP. Mitochondrial dye, λex: 405 nm; λem: 440-500 nm. HSDPP,
λex: 458 nm; Green channel: 500-600 nm; Red channel: 600-700 nm. The scale bar is 10 μm.
SAM-induced mice brains compared to with that
of the saline treated mice. These data
demonstrated the capability of HSDPP in BBB
penetration and tracing the dynamic changes of
H2S levels as well as visualizing the short
emssion was “switched” by mtDNA into red
signals in the mice brains.
within mitochondrial in vitro/vivo, with
multiresponse signals. In particular, it deserved
to be mentioned that, for the first time, we
successfully applied this probe for monitoring
H2S “switched” by mtDNA in living cells and
mapping the endogenous H2S upregulation in
mice brains. In mechanism, as long as the
analytes reacted with the responsive group and
induced the process of H-type aggregates
changing to monomers, the activatable and
switchable probe could realize the consecutive
fluorescence imaging. Therefore, our work not
Conclusion
In conclusion, we proposed a novel design
method towards the first activatable and
switchable probe (HSDPP) for monitoring H2S
and tracking the endogenous H2S upregulation
10
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
only provides a robust chemical tool for probing
endogenous H2S upregulation in vitro/in vivo,
but also affords a forward-looking design
strategy for developing promising versatile or
switchable chemosensors for unveiling the
interrelated roles of biomolecules and mtDNA in
9941.
[5] J. Rao, A. Dragulescu-Andrasi, H. Yao,
Curr. Opin. Biotechnol. 2007, 18, 17–25.
[6] S. Lee, K. Park, K. Kim, K. Choi, I. C.
Kwon, Chem. Commun. 2008, 4250–
4260.
various
physiological
and
pathological
[7] H. Kobayashi, M. Ogawa, R. Alford, P.
L. Choyke, Y. Urano, Chem. Rev. 2010,
110, 2620–2640.
conditions.
Acknowledgements
The authors gratefully acknowledge the financial
support from the Major Science and Technology
Program for Water Pollution Control and
Treatment, China (No. 2017ZX07602-002).
Conflict of interest
[8] Z. Guo, C. Yan, W. Zhu, Angew. Chemie
Int. Ed. 2020, 59, 9812–9825.
[9] Y. Lei, W. Ren, C. K. Wang, R. R. Tao,
H. J. Xiang, L. L. Feng, Y. P. Gao, Q.
Jiang, X. Li, Y. Hu, F. Han, Theranostics
2019, 9, 5672–5680.
The authors declare no competing financial
interest.
[10] Z. Thiel, P. Rivera-Fuentes, Angew.
Chemie - Int. Ed. 2019, 58, 11474–
11478.
Key words: Activatable and switchable
nanoaggregate probe; Hydrogen sulfide; mtDNA;
Mice brains.
[11] J. Cheng, Y. Ren, Y. Huang, X. Li, M.
Huang, F. Han, X. Liang, X. Li, Anal.
Chem. 2020, 92, 1409–1415.
References
[12] L. He, X. Yang, K. Xu, X. Kong, W. Lin,
Chem. Sci. 2017, 8, 6257–6265.
[1] H. Kobayashi, P. L. Choyke, Acc. Chem.
Res. 2011, 44, 83–90.
[13] C. Szabõ, Nat. Rev. Drug Discov. 2007,
6, 917–935.
[2] Y. Lyu, K. Pu, Adv. Sci. 2017, 4,
1600481.
[14] G. Yang, L. Wu, B. Jiang, W. Yang, J.
Qi, K. Cao, Q. Meng, A. K. Mustafa, W.
Mu, S. Zhang, S. H. Snyder, R. Wang,
Science (80-. ). 2008, 322, 587–590.
[3] S. Lee, X. Chen, Mol. Imaging Probes
Cancer Res. 2012, 519–543.
[4] M. Swierczewska, S. Lee, X. Chen, Phys.
Chem. Chem. Phys. 2011, 13, 9929–
11
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
[15] F. Yu, P. Li, P. Song, B. Wang, J. Zhao,
K. Han, Chem. Commun. 2012, 48,
2852–2854.
Zhang, F. Feng, R. Yang, Anal. Chem.
2020, 92, 3555–3562.
[26] L. Yang, J. Zheng, Z. Zou, H. Cai, P. Qi,
Z. Qing, Q. Yan, L. Qiu, W. Tan, R.
Yang, Chem. Commun. 2020, 56, 1843–
1846.
[16] X. Zhang, W. Qu, H. Liu, Y. Ma, L.
Wang, Q. Sun, F. Yu, Anal. Chim. Acta
2020, 1109, 37–43.
[17] F. Yu, X. Han, L. Chen, Chem. Commun.
2014, 50, 12234–12249.
[27] Y. Zhang, S. Yuan, G. Day, X. Wang, X.
Yang, H. C. Zhou, Coord. Chem. Rev.
2018, 354, 28–45.
[18] M. R. Filipovic, J. Zivanovic, B. Alvarez,
R. Banerjee, Chem. Rev. 2018, 118,
1253–1337.
[28] J. M. Estrela, A. Ortega, E. Obrador,
Glutathione in Cancer Biology and
Therapy, 2006.
[19] W. Chen, S. Xu, J. J. Day, D. Wang, M.
Xian, Angew. Chemie - Int. Ed. 2017, 56,
16611–16615.
[29] W. Z. Mehal, I. Inayat, R. A. Flavell,
Science (80-. ). 2006, 847–851.
[30] J. L. Wallace, R. Wang, Nat. Rev. Drug
Discov. 2015, 14, 329–345.
[20] V. S. Lin, W. Chen, M. Xian, C. J.
Chang, Chem. Soc. Rev. 2015, 44, 4596–
4618.
[31] G. C. Kujoth, Science (80-. ). 2005, 309,
481–484.
[21] Y. Luo, C. Zhu, D. Du, Y. Lin, Anal.
Chim. Acta 2019, 1061, 1–12.
[32] S. DiMauro, E. A. Schon, N. Engl. J.
Med. 2003, 348, 2656–2668.
[22] F. Li, Y. Zhou, H. Yin, S. Ai, Biosens.
Bioelectron. 2020, 166, 112476.
[23] J. Mei, Y. Hong, J. W. Y. Lam, A. Qin,
Y. Tang, B. Z. Tang, Adv. Mater. 2014,
26, 5429–5479.
[33] S. V Baudouin, D. Saunders, W.
Tiangyou, J. L. Elson, J. Poynter, A.
Pyle, S. Keers, D. M. Turnbull, N.
Howell, P. F. Chinnery, Lancet 2005,
366, 2118–2121.
[24] C. Chen, Z. Song, X. Zheng, Z. He, B.
Liu, X. Huang, D. Kong, D. Ding, B. Z.
Tang, Chem. Sci. 2017, 8, 2191–2198.
[25] Z. Zou, X. Liao, L. Yang, Z. Huang, H.
Yang, Q. Yan, Y. Zhang, Z. Qing, L.
[34] A. Latorre-Pellicer, R. Moreno-
Loshuertos, A. V. Lechuga-Vieco, F.
Sánchez-Cabo, C. Torroja, R. Acín-
Pérez, E. Calvo, E. Aix, A. González-
12
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
Guerra, A. Logan, M. L. Bernad-Miana,
[42] Y. Liu, J. Nie, J. Niu, W. Wang, W. Lin,
J. Mater. Chem. B 2018, 6, 1973–1983.
[43] X. Zou, Y. Shi, R. Zhu, J. Han, S. Han,
Anal. Chem. 2019, 91, 15899–15907.
[44] M. Tian, J. Sun, B. Dong, W. Lin,
Angew. Chemie - Int. Ed. 2018, 57,
16506–16510.
E. Romanos, R. Cruz, S. Cogliati, B.
Sobrino, Á. Carracedo, A. Pérez-Martos,
P. Fernández-Silva, J. Ruíz-Cabello, M.
P. Murphy, I. Flores, J. Vázquez, J. A.
Enríquez, Nature 2016, 535, 561–565.
[35] K. Shimada, T. R. Crother, J. Karlin, J.
Dagvadorj, N. Chiba, S. Chen, V. K.
Ramanujan, A. J. Wolf, L. Vergnes, D.
M. Ojcius, A. Rentsendorj, M. Vargas, C.
Guerrero, Y. Wang, K. A. Fitzgerald, D.
M. Underhill, T. Town, M. Arditi,
[45] A. C. Sedgwick, L. Wu, H. H. Han, S. D.
Bull, X. P. He, T. D. James, J. L. Sessler,
B. Z. Tang, H. Tian, J. Yoon, Chem. Soc.
Rev. 2018, 47, 8842–8880.
[46] H. Fang, S. Yao, Q. Chen, C. Liu, Y. Cai,
S. Geng, Y. Bai, Z. Tian, A. L. Zacharias,
T. Takebe, Y. Chen, Z. Guo, W. He, J.
Diao, ACS Nano 2019, 13, 14426–14436.
[47] F. Würthner, T. E. Kaiser, C. R. Saha-
Möller, Angew. Chemie - Int. Ed. 2011,
50, 3376–3410.
Immunity 2012, 36, 401–414.
[36] A. P. West, G. S. Shadel, Nat. Rev.
Immunol. 2017, 17, 363–375.
[37] R. H. Swerdlow, S. M. Khan, Med.
Hypotheses 2004, 63, 8–20.
[38] R. H. Swerdlow, J. M. Burns, S. M.
Khan, J. Alzheimer’s Dis. 2010, 20, DOI
10.3233/JAD-2010-100339.
[48] H. Zhu, C. Liang, X. Cai, H. Zhang, C.
Liu, P. Jia, Z. Li, Y. Yu, X. Zhang, W.
Sheng, B. Zhu, Anal. Chem. 2020,
acs.analchem.9b04009.
[39] R. K. Chaturvedi, M. F. Beal, Free
Radic. Biol. Med. 2013, 63, 1–29.
[40] Z. Chen, X. Mu, Z. Han, S. Yang, C.
Zhang, Z. Guo, Y. Bai, W. He, J. Am.
Chem. Soc. 2019, jacs.9b09181.
[49] Y. Wang, L. Zhang, S. Zhang, Z. Liu, L.
Chen, Anal. Chem. 2019, 91, 7812–7818.
[50] J. Bin Li, H. W. Liu, T. Fu, R. Wang, X.
B. Zhang, W. Tan, Trends Chem. 2019,
1, 224–234.
[41] C. J. Reinhardt, E. Y. Zhou, M. D.
Jorgensen, G. Partipilo, J. Chan, J. Am.
Chem. Soc. 2018, 140, 1011–1018.
13
This article is protected by copyright. All rights reserved.

10.1002/asia.202000964
Chemistry - An Asian Journal
A novel activatable and switchable nanoaggregate probe for detecting H2S in mice brains was
developed. It could be effectively activated by H2S to form H-type aggregates with green emission;
subsequently, the aggregates could bind to mtDNA to form monomers and the emssion color switched
from green to deep-red.
14
This article is protected by copyright. All rights reserved.