Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 322 (2024) 124810

Contents lists available at ScienceDirect

Spectrochimica Acta Part A:

Molecular and Biomolecular Spectroscopy
journal homepage: www.journals.elsevier.com/spectrochimica-acta-part-a-
molecular-and-biomolecular-spectroscopy

A Golgi-targeted fluorescent probe for monitoring polarity dynamic during

programmed cell death
Feiran Liu a , Zichun Li a , Jing Jing a , Xiaoling Zhang a, b
a
Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering, Ministry of Industry and Information Technology, Key Laboratory of Cluster Science of
Ministry of Education, Beijing Key Laboratory of Photo-electronic/Electrophotonic Conversion Materials, Analytical and Testing Center, School of Chemistry and
Chemical Engineering, Beijing Institute of Technology, Beijing, 100081, PR China
b
School of Medical Technology, Beijing Institute of Technology, Beijing, 100081, PR China

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• GTO was designed for monitoring

polarity. A Golgi apparatus-targeted fluorescent probe has been successfully developed for monitoring polarity
• GTO could be localized to Golgi dynamic during programmed cell death.
apparatus.
• GTO was able to monitor polarity dy­
namic during the early event of pro­
grammed cell death.

A R T I C L E I N F O A B S T R A C T

Keywords: Programmed cell death (PCD) is a controlled form of cell death and it plays an essential role in maintaining
Golgi Apparatus homeostasis. Golgi apparatus works as the hotspot during the early event of PCD and Golgi polarity, a vital
Fluorescent Probe microenvironment factor, can be regarded as an indicator of physiological status. Combined Golgi-targeted group
Polarity
phenylsulfonamide as electron acceptor group and triphenylamine as electron donor group, a novel Golgi-
Programmed Cell Death
targeted fluorescent probe GTO had been developed. GTO showed good sensitivity and selectivity to polarity
and its remarkable photostability makes it potentially useful for long-term cellular monitoring. In practice, GTO
demonstrated good cell permeability and Golgi targeting capabilities. According to our results, GTO was applied
to reveal the polarity increase during the early event of PCD and the encouraging results illustrated that GTO was
an imaging tool for monitoring polarity in Golgi apparatus and the exploration in early diagnosis and drug
discovery.

E-mail addresses: [email protected] (J. Jing), [email protected] (X. Zhang).

https://doi.org/10.1016/j.saa.2024.124810
Received 8 April 2024; Received in revised form 5 July 2024; Accepted 10 July 2024
Available online 10 July 2024
1386-1425/© 2024 Elsevier B.V. All rights are reserved, including those for text and data mining, AI training, and similar technologies.
F. Liu et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 322 (2024) 124810

Scheme 1. Synthetic procedure of GTO.

1. Introduction showed excellent sensitivity and selectivity to polarity. Moreover, its

remarkable photostability makes it potentially useful for long-term
Programmed cell death (PCD) is a controlled form of cell death. It is cellular monitoring. Besides, the cells experiment verified the capacity
precisely controlled by a series of molecular events which are activated of GTO to monitor polarity dynamic within Golgi apparatus. Moreover,
by various physiological or developmental signals. As a vital component, GTO was applied to monitor polarity dynamic during the early event of
PCD plays an essential role in maintaining tissue homeostasis by PCD and reveal the increase of polarity during those processes. The
removing nonfunctional, damaged and harmful cells. With the in-depth encouraging result indicated the great potential of GTO for further
study, more types of PCD, including apoptosis, ferroptosis, cuproptosis, application in early diagnosis and drug discovery.
have been identified. However, more research should be done to clearly
find out the complex regulatory mechanism that governs programmed 2. Experiments
cell death [1–7].
Golgi apparatus is the central for the post-translational modification 2.1. Materials and instruments
and sorting. After received cargo from endoplasmic reticulum, lipids and
proteins will be modified first, then sorted and packed. Finally, the cargo The reagents and solvents used for all experiments were purchased
will be transported to their appropriate destination [8–11]. As a result of from commercial sources and used without further purification. 1H NMR
this, the homeostasis in Golgi apparatus is important to various cellular and 13C NMR spectra of GTO were recorded by a Bruker Avance III 400
processes, and the dysfunction of Golgi affects multiple cell functions, MHz spectrometer. Bruker Apex IV Fourier was used for taking High-
which may finally cause various diseases [12–15]. Likewise, Golgi resolution mass spectra (HRMS). In optical study, a Purkinje TU-1901
apparatus plays an essential role in ensuring PCD processes normally. spectrophotometer was used to record the absorption spectrum, and a
Previous studies have shown that Golgi apparatus participates in the Hitachi F-7000 fluorescence spectrometer was used to measure the
induction of PCD, and some Golgi-residing proteins are involved in the fluorescence spectrum, respectively. The pH of the test solution was
signaling and execution of PCD [16–20]. Although we have already determined by FE-30 pH meter made by Mettler Toledo. Fluorescence
known that Golgi apparatus can sense and integrate diversified death images of cells were taken by Olympus FV1000 confocal fluorescence
signals, it still need time to investigate the connection between PCD and microscope.
Golgi apparatus more clearly.
Polarity, as a vital microenvironment factor, can be regard as an
indicator of physiological status, because the normal operation of 2.2. Synthetic procedure of GTO
various cellular processes, including protein transportation, enzyme
catalysis and signal transduction, relies on the homeostasis of polarity. Scheme 1 described the synthetic procedure of GTO. In a round-
To refer in particular to Golgi apparatus, the maintenance of polarity bottom flask, 4-bromo-N,N-bis(4-methoxyphenyl)aniline (384 mg, 1
homeostasis is important to keep this crucial organelle operating regu­ mmol), (4-Aminosulfonylphenyl)boronic acid (201 mg, 1 mmol) and
larly [21–24]. Besides, considering its essential role in transduction and (Ph3P)4Pd (58 mg, 0.05 mmol) was added with a stirring bar. Then,
transportation, Golgi apparatus works as the hotspot during the early K2CO3 aqueous solution (1.5 mL, 2 mol/L, 3 mmol) and 10 mL THF was
event of various cellular processes, including PCD [19,20,25–27]. added. The system was heated to 70 ◦ C and stirred overnight under N2.
Hence, monitoring the polarity dynamic within Golgi apparatus during Then, the reactant was evaporated under vacuum and the crude product
the early phase may provide us with early recognition and deep un­ was purified by silica gel column chromatography (DCM/MeOH =
derstanding of the trigger and mechanism of PCD. 100:1, v/v) to afford GTO as yellow solid. Yield: 179 mg, 39 %. ESI-MS:
Nowadays, some methods have been developed for the exploration m/z calcd for C26H24N2O4S: 461.1457, found [C26H25N2O4S]+:
of Golgi apparatus, but there are still some limitations on those existing 461.1515. 1H NMR (400 MHz, DMSO‑d6) δ (ppm): 7.91 (d, J = 8.2 Hz,
methods in the aspects of Golgi-targeted ability and in situ analysis. 2H), 7.77 (d, J = 8.2 Hz, 2H), 7.56 (d, J = 8.5 Hz, 2H), 7.42 (s, 2H), 7.11
Compared with other methods, fluorescent probe is a good choice to – 7.02 (m, 4H), 6.95 – 6.89 (m, 4H), 6.87 (d, J = 8.7 Hz, 2H), 3.74 (s,
meet the requirement of in situ analysis ability and high sensitivity. 6H). 13C NMR (101 MHz, DMSO) δ (ppm): 156.54, 149.25, 143.47,
Hence, it can be regarded as an effective method to clarify the function 142.31, 140.14, 129.97, 129.58, 128.14, 127.64, 127.58, 127.51,
of Golgi apparatus during various physiological processes comprehen­ 126.72, 126.47, 119.38, 115.53, 55.74.
sively [28–36]. Although some Golgi-targeted fluorescent probes have
been reported for different bioactive species, few polarity-sensitive
2.3. Spectral experiments
Golgi-targeted fluorescent probe was developed [37–40].
Herein, a new Golgi-targeted polarity fluorescent probe GTO was
In vitro experiments, the concentration of GTO was 10 μM, which
constructed. Golgi-targeted group phenylsulfonamide worked as elec­
was diluted from 1 mM stock solution in DMSO. PBS buffer used in the
tron acceptor group and triphenylamine worked as electron donor
experiments was 10 mM. Setting 405 nm as the excitation wavelength.
group, the combination of those two parts formed GTO, a typical D-π-A
The excited slit widths were both 5 nm, which was same as the emission
molecular. Based on the intramolecular charge transfer process, GTO
slit width, unless otherwise stated.

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F. Liu et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 322 (2024) 124810

Fig. 1. (a) The UV–vis spectra of GTO in different solutions; (b) The emission spectra of GTO in different solutions; (c) The emission spectra of GTO in THF-water
systems with different ratios; (d) The fluorescence intensity of GTO at 496 nm in THF-water systems with different ratios compared to △f with error bar.

2.4. Cells-related experiments illustrated in Fig. 1b, with the polarity of the solution increased, the
fluorescence decreased significantly. The results indicated that GTO had
The nutrient solution for cells culturing is 10 % calf serum or foetal a polarity-dependent emission and the potential to act as a polarity in­
bovine serum in Dulbecco’s modified eagle medium (DMEM), with 50 dicator. To verify its optical response accurately, we investigated the
μg/mL penicillin and 50 μg/mL streptomycin. Cells were cultured in a fluorescence of GTO in a series of THF-water systems. The polarity of the
37 ◦ Cincubator involving 5 % CO2. Cells used for imaging were prepared solution system could be adjusted by changing the ratio of THF-water.
by following the process, in the same condition, cells were seeded into As shown in Fig. 1c and d, GTO exhibited high sensitivity to the
confocal dish and cultured before imaging. Before confocal imaging, PBS mixed system with different polarity. Besides, the fluorescence intensity
was used to wash out dead cells and the procedure redid for 3 times. at 496 nm, which showed decreased with the increase of polarity in
To demonstrated the cytotoxicity of GTO, 96-well plates were first parallel experiments (compared to △f), could be used as a signal for
seeded by MCF-7 cells with 3 × 103 cells per well and the plates would analyzing the polarity change as Fig. 1d shown. The above encouraging
use after 24 h incubation in the same condition. Next, added 0, 5, 10, 15 results showed the capacity of GTO, which could be regarded as a
and 20 μM GTO to those wells, cultured for 24 h in the same condition remarkable fluorescent probe to indicate the polarity with high
except the concentrations of the probe, respectively. Finally, in each sensitivity.
well, added 10 μL CCK-8 solution, incubated 4 h. After the above pro­ The selectivity of GTO to polarity had been estimated. First, as shown
cess, by using a Thermo Scientific Multiskan FC, the OD was recorded to in Figure S2, the intensity of fluorescence hadn’t been affected by the
be calculated. addition of some common interfering substances, which performed the
great selectivity of GTO to polarity. Besides, the influence of other
3. Results and discussion microenvironment parameters on GTO had also been estimated. On one
hand, as shown in Figure S3A, the intensity was basically unchanged by
3.1. Optical responses pH in the range of Golgi apparatus. On the other hand, compared to low
polarity microenvironment, the fluorescence intensity change induced
To begin with, the optical response of GTO to polarity is assessed. by viscosity was negligible (Figure S3B). Therefore, the result demon­
GTO was first induced into different solutions to investigate its ab­ strated the capacity for GTO to monitor the polarity without being
sorption and fluorescence. According to Fig. 1a, the absorption peaks of affected by other substances. In addition, even irradiated by Xenon lamp
GTO in different solutions are basically consistent. Considering the for 30 min, the fluorescence of GTO was still stable and without any
shortest wavelength on the confocal microscope equipped was 405 nm, changes, which indicated the good photostability of GTO (Figure S4).
we selected 405 nm as the excitation to keep the consistent condition
both in vitro and in vivo (Figure S1). Setting 405 nm as the excitation
wavelength, the fluorescence spectra of GTO had been recorded. As

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F. Liu et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 322 (2024) 124810

Fig. 2. Co-location of GTO and Golgi-Tracker Red. MCF-7 cells were treated with GTO and Golgi-Tracker Red. (a) GTO channel, Ch1 (λex = 405 nm, λem = 450–530
nm); (b) Golgi-Tracker Red, Ch2, (λex = 543 nm, λem = 550–650 nm); (c) Brightfield; (d) Coverage of a-c graphs; (e) Correlation graph of fluorescence intensity of Ch1
and Ch2 channels; (f) The intensity curve of two channels in the straight line 1.

3.2. The imaging capability of GTO within Golgi apparatus fluorescence from GTO didn’t show any decrease, which verified the
good photostability of GTO and the potential to monitor polarity dy­
Before being applied in vivo, the cytotoxicity, subcellular localiza­ namic within Golgi apparatus.
tion and photostability of GTO have been examined. First, in the aspect
of cytotoxicity, according to the results demonstrated in Figure S5. GTO 3.3. Fluorescence imaging of Golgi polarity change in various cellular
showed negligible cytotoxicity to MCF-7 cells in 5–20 μM, which indi­ stress processes
cated that GTO was suited for cell imaging. Next, by using the com­
mercial targeting probe Golgi-Tracker Red, the subcellular localization Cellular stress is mainly about the situation when cells are exposed to
of GTO has been assessed. As shown in Fig. 2, there was a good overlap an adverse environmental condition that significantly perturbs cellular
between the fluorescence from the tracker and GTO. The Pearson co­ homeostasis. Cellular stress, including Endoplasmic Reticulum Stress,
efficient is 0.93, which indicated that GTO was able to target Golgi Golgi stress, thermal stress, hypoxia and oxidative stress, plays an
apparatus. To end with, the photostability of GTO has been evaluated by essential role in kinds of cellular processes [41,42]. Setting Golgi stress
multiple scanning under the same condition as cell imaging. As and ER stress as examples, we investigated the polarity change during
demonstrated in Figure S6, under the irradiation of 405 nm laser, the those cellular stress and verified the capacity of GTO to recognize

Fig. 3. (A)Fluorescence imaging of Golgi polarity in MCF-7 cells. (a–c) MCF-7 cells only treated with GTO (10 μM); (d–f) MCF-7 cells pre-treated with 10 μM
Monensin for 6 h and then treated with GTO (10 μM); (g–i) MCF-7 cells pre-treated with 10 μM Tunicamycin for 1 h and then treated with GTO (10 μM). (B) a, d, g
channels’ intensity. λex = 405 nm, λem = 450–550 nm. Scale bar: 30 μm.

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F. Liu et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 322 (2024) 124810

Fig. 4. (A) Fluorescence imaging of Golgi polarity in MCF-7 cells during the early events in Apoptosis. MCF-7 cells were pre-treated with GTO (10 μM) and then
treated with dexamethasone (10 μM) for (a–c) 0 min, (d–f) 15 min, (g–i) 30 min, (j–l) 45 min, (m–o) 60 min. (B) a, d, g, j, m channels’ intensity. λex = 405 nm, λem =
450–550 nm. Scale bar: 30 μm.

polarity change within Golgi apparatus. (Figure S7).

Golgi stress is accompanied by the disruption of redox balance within
Golgi apparatus, which may induce a variety of diseases [43,44].
3.4. Monitoring polarity dynamic during programmed cell death within
Monensin is a carboxylic ionophore which can bind Na+/H+. According
Golgi apparatus
to previous reports, Monensin can specifically induce Golgi Stress [45].
GTO was applied to investigate polarity change within Golgi apparatus
Based on the main function of Golgi apparatus, it can be inferred that
by using Monensin to specifically induce Golgi stress in MCF-7 cells.
Golgi apparatus plays as the central of signaling pathways to regulate
According to Fig. 3, cells which have been pre-treated with Monensin for
PCD to proceed normally. As a result, Golgi apparatus may play an
6 h were incubated with GTO. The fluorescence showed a significant
essential role in the early event of PCD. Hence, by using the important
decrease compared to the blank group, which could indicate that the
microenvironment factor polarity as the indicator, real-time continuous
polarity within Golgi apparatus was increased during Golgi stress.
monitoring polarity within Golgi apparatus during PCD has great po­
As an important process for regulating protein folding and traf­
tential for early diagnosis and screening drugs. Herein, set Apoptosis and
ficking, ER Stress has a strong connection with Golgi apparatus [46,47].
Ferroptosis as examples, GTO was applied to monitor the polarity dy­
We constructed the experiments to investigate the influence of ER stress
namic within Golgi apparatus during the early event of PCD.
on Golgi polarity by pre-treated cells with Tunicamycin, which is a well-
As one of the most common PCD processes, Apoptosis is important in
known ER stressor by disrupting protein processing and folding [48]. As
the maintenance of homeostasis and its abnormity induces a variety of
illustrated in Fig. 3, cells were first treated with Tunicamycin and then
diseases [19]. Although several studies have demonstrated that Golgi
treated with GTO (10 μM), which were showed weak fluorescence. The
apparatus disassembled during Apoptosis, experiment results showed
decreased fluorescence intensity showed the polarity increase within
that Golgi apparatus played a crucial role in sensing and regulating early
Golgi apparatus during ER stress.
events in Apoptosis [26,27]. Thus, GTO was used to investigate the
Further, HT-22 cells were selected to demonstrate the performance
polarity dynamic within Golgi apparatus during early events in
of GTO in normal tissue cells. After the treatment of Monensin or
Apoptosis. As shown in Fig. 4, cells were treated with Dexamethasone
Tunicamycin to induce cellular stress, the fluorescence of GTO showed a
(10 μM) to induce Apoptosis, and the fluorescence decreased with the
decrease compared to the blank group, which indicated the increase of
incubating time (0–60 min) increase. The results revealed the polarity
Golgi polarity during cellular stress, and verified GTO’s sensitive
increases during early events in apoptosis.
response to polarity and its potential utilization in different cell lines
As an iron-dependent process, Ferroptosis induces excessive

Fig. 5. (A) Fluorescence imaging of Golgi polarity in MCF-7 cells during the early events in Ferroptosis. MCF-7 cells were pre-treated with GTO (10 μM) and then
treated with Erastin (10 μM) for (a–c) 0 min, (d–f) 15 min, (g–i) 30 min, (j–l) 45 min, (m–o) 60 min. (B) a, d, g, j, m channels’ intensity. λex = 405 nm, λem = 450–550
nm. Scale bar: 30 μm.

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accumulation of lipid peroxidation and is involved in various diseases. References

According to previous research, pieces of evidence showed that Golgi
apparatus might play an essential role during Ferroptosis [20,25]. [1] W. Park, S. Wei, B. Kim, B. Kim, S. Bae, Y. Chae, D. Ryu, K. Ha, Diversity and
complexity of cell death: a historical review, Exp. Mol. Med. 55 (2023) 1573–1594,
However, there is a need for more research to figure out the exact https://doi.org/10.1038/s12276-023-01078-x.
mechanism between Golgi apparatus and Ferroptosis. Hence, GTO was [2] D. Tang, R. Kang, T.V. Berghe, P. Vandenabeele, G. Kroemer, The molecular
applied to monitor the polarity dynamic within Golgi apparatus during machinery of regulated cell death, Cell Res. 29 (2019) 347–364, https://doi.org/
10.1038/s41422-019-0164-5.
Ferroptosis. Erastin, a well-known Ferroptosis inducer, was used to [3] J. Yuan, D. Ofengeim, A guide to cell death pathways, Nat. Rev. Mol. Cell Biol. 25
trigger Ferroptosis without the apparent of other types of PCD [49]. As (2024) 379–395, https://doi.org/10.1038/s41580-023-00689-6.
illustrated in Fig. 5, with the increase of incubating time (0–60 min), the [4] D. Bertheloot, E. Latz, B.S. Franklin, Necroptosis, pyroptosis and apoptosis: an
intricate game of cell death, Cell. Mol. Immunol. 18 (2021) 1106–1121, https://
fluorescence intensity of GTO showed a significant decrease, which doi.org/10.1038/s41423-020-00630-3.
could be indicated as the increase of polarity within Golgi apparatus [5] G. Lei, L. Zhuang, B. Gan, Targeting ferroptosis as a vulnerability in cancer, Nat.
during Ferroptosis. Rev. Cancer 22 (2022) 381–396, https://doi.org/10.1038/s41568-022-00459-0.
[6] P. Tsvetkov, S. Coy, B. Petrova, M. Dreishpoon, A. Verma, M. Abdusamad,
Similar experiments were done on HT-22 cells to monitor Golgi po­
J. Rossen, L. Joesch-Cohen, R. Humeidi, R.D. Spangler, J.K. Eaton, E. Frenkel,
larity dynamic during PCD on normal tissue cells. As shown in Figure S8 M. Kocak, S.M. Corsello, S. Lutsenko, N. Kanarek, S. Santagata, T.R. Golub, Copper
& S9, the fluorescence of GTO realized the visualization of Golgi polarity induces cell death by targeting lipoylated TCA cycle proteins, Science 375 (2022)
dynamic during the early stage of Apoptosis and Ferroptosis, the obvious 1254–1261, https://doi.org/10.1126/science.abf0529.
[7] P. Yu, X. Zhang, N. Liu, L. Tang, C. Peng, X. Chen, Pyroptosis: mechanisms and
decrease of fluorescence intensity indicated the polarity increase within diseases, Signal Transduct. Target. Ther. 6 (2021) 128, https://doi.org/10.1038/
Golgi apparatus in those processes. Those encouraging results demon­ s41392-021-00507-5.
strate the excellent performance of the probe GTO to work as an effec­ [8] M.A. De Matteis, A. Luini, Exiting the Golgi complex, Nat. Rev. Mol. Cell Biol. 9
(2008) 273–284, https://doi.org/10.1038/nrm2378.
tive tool for visualizing polarity (Table S1). [9] M. Lowe, Structural organization of the Golgi apparatus, Curr. Opin. Cell Biol. 23
(2011) 85–93, https://doi.org/10.1016/j.ceb.2010.10.004.
4. Conclusion [10] J. Hellicar, N.L. Stevenson, D.J. Stephens, M. Lowe, Supply chain logistics – the
role of the Golgi complex in extracellular matrix production and maintenance,
J. Cell Sci. 135 (2022) jcs258879, https://doi.org/10.1242/jcs.258879.
To sum up, a Golgi-targeted fluorescent probe GTO, which combined [11] I. Agliarulo, S. Parashuraman, Golgi apparatus regulates plasma membrane
Golgi-targeted group phenylsulfonamide as electron acceptor group and composition and function, Cells 11 (2022) 368, https://doi.org/10.3390/
cells11030368.
triphenylamine as electron donor group, had been synthesized. This [12] J. Liu, Y. Huang, T. Li, Z. Jiang, L. Zeng, Z. Hu, The role of the Golgi apparatus in
typical D-π-A molecular showed high photostability, excellent sensi­ disease (Review), Int. J. Mol. Med. 47 (2021) 38, https://doi.org/10.3892/
tivity and selectivity to polarity. Besides, GTO was applied to monitor ijmm.2021.4871.
[13] M.G. Bexiga, J.C. Simpson, Human diseases associated with form and function of
polarity dynamic during the early event of PCD and reveal the increase
the Golgi complex, Int. J. Mol. Sci. 14 (2013) 18670–18681, https://doi.org/
of Golgi polarity during those processes. The encouraging result indi­ 10.3390/ijms140918670.
cated that GTO can be used for monitoring polarity in Golgi apparatus [14] S. Potelle, A. Klein, F. Foulquier, Golgi post-translational modifications and
and further application in early diagnosis. associated diseases, J. Inherit. Metab. Dis. 38 (2015) 741–751, https://doi.org/
10.1007/s10545-015-9851-7.
[15] J. Li, Y. Wang, Golgi metal ion homeostasis in human health and diseases, Cells 11
CRediT authorship contribution statement (2022) 289, https://doi.org/10.3390/cells11020289.
[16] R.S. Maag, S.W. Hicks, C.E. Machamer, Death from within: apoptosis and the
secretory pathway, Curr. Opin. Cell Biol. 15 (2003) 456–461, https://doi.org/
Feiran Liu: Writing – original draft, Visualization, Validation, 10.1016/S0955-0674(03)00075-9.
Methodology, Conceptualization. Zichun Li: Methodology, Conceptu­ [17] B.J. van Raam, T. Lacina, R.K. Lindemann, J.H. Reiling, Secretory stressors induce
alization. Jing Jing: Writing – review & editing, Supervision, Resources. intracellular death receptor accumulation to control apoptosis, Cell Death Dis 8
(2017) e3069, https://doi.org/10.1038/cddis.2017.466.
Xiaoling Zhang: Writing – review & editing, Supervision, Resources, [18] D. Wlodkowic, J. Skommer, D. McGuinness, C. Hillier, Z. Darzynkiewicz, ER–Golgi
Funding acquisition. network—a future target for anti-cancer therapy, Leuk. Res. 33 (2009) 1440–1447,
https://doi.org/10.1016/j.leukres.2009.05.025.
[19] Y.A. Hannun, L.M. Obeid, Sphingolipids and their metabolism in physiology and
Declaration of competing interest disease, Nat. Rev. Mol. Cell Biol. 19 (2018) 175–191, https://doi.org/10.1038/
nrm.2017.107.
The authors declare that they have no known competing financial [20] H. Alborzinia, T.I. Ignashkova, F.R. Dejure, M. Gendarme, J. Theobald, S. Wölfl, R.
K. Lindemann, J.H. Reiling, Golgi stress mediates redox imbalance and ferroptosis
interests or personal relationships that could have appeared to influence
in human cells, Commun. Biol. 1 (2018) 210, https://doi.org/10.1038/s42003-
the work reported in this paper. 018-0212-6.
[21] J. Yin, L. Huang, L. Wu, J. Li, T.D. James, W. Lin, Small molecule based fluorescent
Data availability chemosensors for imaging the microenvironment within specific cellular regions,
Chem. Soc. Rev. 50 (2021) 12098–12150, https://doi.org/10.1039/D1CS00645B.
[22] H. Xiao, P. Li, B. Tang, Recent progresses in fluorescent probes for detection of
Data will be made available on request. polarity, Coord. Chem. Rev. 427 (2021) 213582, https://doi.org/10.1016/j.
ccr.2020.213582.
[23] H.R. Petty, Fluorescence microscopy: established and emerging methods,
Acknowledgments experimental strategies, and applications in immunology, Microsc. Res. Tech. 70
(2007) 687–709, https://doi.org/10.1002/jemt.20455.
The work was done with the support of the National Natural Science [24] Y. Ravichandran, B. Goud, J. Manneville, The Golgi apparatus and cell polarity:
Roles of the cytoskeleton, the Golgi matrix, and Golgi membranes, Curr. Opin. Cell
Foundation of China Project (No.22174008, 22177013, 21974009 and Biol. 62 (2020) 104–113, https://doi.org/10.1016/j.ceb.2019.10.003.
22274010). Thanks for the support of the Analysis and Testing Center of [25] X. Chen, R. Kang, G. Kroemer, D. Tang, Organelle-specific regulation of ferroptosis,
Beijing Institute of Technology. Cell Death Differ. 28 (2021) 2843–2856, https://doi.org/10.1038/s41418-021-
00859-z.
[26] C.E. Machamer, Golgi disassembly in apoptosis: cause or effect? Trends Cell Biol.
Appendix A. Supplementary material 13 (2003) 279–281, https://doi.org/10.1016/S0962-8924(03)00101-6.
[27] S. Mukherjee, R. Chiu, S. Leung, D. Shields, Fragmentation of the Golgi apparatus:
an early apoptotic event independent of the cytoskeleton, Traffic 8 (2007)
Supplementary data to this article can be found online at https://doi.
369–378, https://doi.org/10.1111/j.1600-0854.2007.00542.x.
org/10.1016/j.saa.2024.124810. [28] R. Li, C. Wen, C. Huang, N. Li, Functional molecules and nano-materials for the
Golgi apparatus-targeted imaging and therapy, TrAC Trends Anal. Chem. 156
(2022) 116714, https://doi.org/10.1016/j.trac.2022.116714.
[29] C. Liu, H. Zhu, Y. Zhang, M. Su, M. Liu, X. Zhang, X. Wang, X. Rong, K. Wang, X. Li,
B. Zhu, Recent advances in Golgi-targeted small-molecule fluorescent probes,

6
F. Liu et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 322 (2024) 124810

Coord. Chem. Rev. 462 (2022) 214504, https://doi.org/10.1016/j. [39] X. Feng, G. Zhang, R. Sun, Y. Xu, J. Ge, Golgi polarity fluorescent imaging based on
ccr.2022.214504. coumarin or 1,8-naphthalimide derivatives in three channels, Sens. Actuators B
[30] H. Zhang, J. Fan, J. Wang, S. Zhang, B. Dou, X. Peng, An off–on COX-2-specific 394 (2023) 134469, https://doi.org/10.1016/j.snb.2023.134469.
fluorescent probe: targeting the Golgi apparatus of cancer cells, J. Am. Chem. Soc. [40] X. Wang, X. Li, Y. Zhou, S. Wei, Y. Li, B. Fan, C. Jia, H. Wang, B. Xue, A golgi-
135 (2013) 11663–11669, https://doi.org/10.1021/ja4056905. targeting and polarity-specific fluorescent probe for the diagnosis of cancer and
[31] H. Wang, Z. He, Y. Yang, J. Zhang, W. Zhang, W. Zhang, P. Li, B. Tang, Ratiometric fatty liver in living cells and tissues, Talanta 268 (2024) 125367, https://doi.org/
fluorescence imaging of Golgi H2O2 reveals a correlation between Golgi oxidative 10.1016/j.talanta.2023.125367.
stress and hypertension, Chem. Sci. 10 (2019) 10876–10880, https://doi.org/ [41] D. Kültz, Evolution of the cellular stress proteome: from monophyletic origin to
10.1039/C9SC04384E. ubiquitous function, J. Exp. Biol. 206 (2003) 3119–3124, https://doi.org/
[32] H. Zhu, C. Liang, X. Cai, H. Zhang, C. Liu, P. Jia, Z. Li, Y. Yu, X. Zhang, W. Sheng, 10.1242/jeb.00549.
B. Zhu, Rational design of a targetable fluorescent probe for visualizing H2S [42] D. Kültz, Molecular and evolutionary basis of the cellular stress response, Annu.
production under Golgi stress response elicited by Monensin, Anal. Chem. 92 Rev. Physiol. 67 (2004) 225–257, https://doi.org/10.1146/annurev.
(2020) 1883–1889, https://doi.org/10.1021/acs.analchem.9b04009. physiol.67.040403.103635.
[33] Z. Zheng, S. Feng, S. Gong, G. Feng, Golgi-targetable fluorescent probe for [43] J.I. Sbodio, S.H. Snyder, B.D. Paul, Golgi stress response reprograms cysteine
ratiometric imaging of CO in cells and zebrafish, Sens. Actuators B 347 (2021) metabolism to confer cytoprotection in Huntington’s disease, Proc. Natl. Acad. Sci.
130631, https://doi.org/10.1016/j.snb.2021.130631. 115 (2018) 780–785, https://doi.org/10.1073/pnas.1717877115.
[34] Z. He, D. Liu, Y. Liu, X. Li, W. Shi, H. Ma, Golgi-targeted fluorescent probe for [44] J.I. Sbodio, B.D. Paul, C.E. Machamer, S.H. Snyder, Golgi protein ACBD3 mediates
imaging NO in Alzheimer’s disease, Anal. Chem. 94 (2022) 10256–10262, https:// neurotoxicity associated with Huntington’s disease, Cell Rep. 4 (2013) 890–897,
doi.org/10.1021/acs.analchem.2c01885. https://doi.org/10.1016/j.celrep.2013.08.001.
[35] Z. Tang, Z. Yan, L. Gong, L. Zhang, X. Yin, J. Sun, K. Wu, W. Yang, G. Fan, Y. Li, [45] M. Oku, S. Tanakura, A. Uemura, M. Sohda, Y. Misumi, M. Taniguchi,
H. Jiang, Precise monitoring and assessing treatment response of sepsis-induced S. Wakabayashi, H. Yoshida, Novel cis-acting element GASE regulates
acute lung hypoxia with a nitroreductase-activated Golgi-targetable fluorescent transcriptional induction by the Golgi stress response, Cell Struct. Funct. 36 (2011)
probe, Anal. Chem. 94 (2022) 14778–14784, https://doi.org/10.1021/acs. 1–12, https://doi.org/10.1247/csf.10014.
analchem.2c03722. [46] C.M. Gallagher, P. Walter, Ceapins inhibit ATF6α signaling by selectively
[36] S. Han, H. Wang, H. Li, K. Li, J. Wang, X. Song, Construction of a dual-functional, preventing transport of ATF6α to the Golgi apparatus during ER stress, eLife 5
reversible, ratiometric, Golgi-targeting fluorescent probe for real-time monitoring (2016) e11880, https://doi.org/10.7554/eLife.11880.
of dynamics of intracellular redox homeostasis, Anal. Chem. 95 (2023) 8002–8010, [47] J. Shen, X. Chen, L. Hendershot, R. Prywes, ER stress regulation of ATF6
https://doi.org/10.1021/acs.analchem.3c00825. localization by dissociation of BiP/GRP78 binding and unmasking of Golgi
[37] P. Li, X. Guo, X. Bai, X. Wang, Q. Ding, W. Zhang, W. Zhang, B. Tang, Golgi localization signals, Dev. Cell 3 (2002) 99–111, https://doi.org/10.1016/S1534-
apparatus polarity indicates depression-like behaviors of mice using in vivo 5807(02)00203-4.
fluorescence imaging, Anal. Chem. 91 (2019) 3382–3388, https://doi.org/ [48] V. Cóppola-Segovia, C. Cavarsan, F.G. Maia, A.C. Ferraz, L.S. Nakao, M.M.S. Lima,
10.1021/acs.analchem.8b04703. S.M. Zanata, ER stress induced by tunicamycin triggers α-synuclein
[38] H. Wang, M. Dong, H. Wang, F. Huang, P. Li, W. Zhang, W. Zhang, B. Tang, oligomerization, dopaminergic neurons death and locomotor impairment: a new
Ultrasensitive and ratiometric two-photon fluorescence imaging of Golgi polarity model of Parkinson’s disease, Mol. Neurobiol. 54 (2017) 5798–5806, https://doi.
during drug-induced acute kidney injury, Chem. Commun. 57 (2021) 5838–5841, org/10.1007/s12035-016-0114-x.
https://doi.org/10.1039/D1CC01411K. [49] X. Li, P. Leung, Erastin-induced ferroptosis is a regulator for the growth and
function of human pancreatic islet-like cell clusters, Cell Regeneration 9 (2020) 16,
https://doi.org/10.1186/s13619-020-00055-3.