mSphere
·
2022
·Vol. 7
·期 4
·
IF 5.029
Hui Li,Cheng-Wen He,Jing Zhu,Zhiping Xie
Genetically encoded tags, such as engineered ascorbate peroxidase APEX2, offer unique advantages for the specific labeling of subcellular structures in electron microscopy (EM). However, the use of APEX2 in EM investigation of yeast has been limited. Here we describe the development of APEX2-based organelle markers for Saccharomyces cerevisiae. We found that with regard to APEX2 -catalyzed formation of diaminobenzidine precipitation, cell wall removal was not essential during sample preparation, yet the presence of fluorescent proteins in APEX2 chimeras had a negative impact. We showed that major organelles including endoplasmic reticulum, early Golgi, late Golgi/early endosomes, late endosomes, mitochondria, peroxisomes, and lipid droplets could be labeled by appropriate APEX2 chimeras. The subcellular localization of our APEX2 chimeras was verified by EM visualization and supplemented with immunofluorescence colocalization analysis when necessary, validating their feasibility as organelle markers.IMPORTANCE Yeast is an excellent single cellular model system for studying basic cellular processes. However, yeast cells are much smaller than most animal and plant cells, making the observation and recognition of yeast subcellular structures challenging. Here we developed a set of yeast organelle markers for use in electron microscopy and documented our technical approach for using this method.
研究领域:
细胞生物学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
Acupuncture and Herbal Medicine
·
2024
·Vol. 4
·期 2
·243
·
IF 0
Wu Jianzhi,Ma Zhi,Zhang Yinhao,Duan Shuni,Sun Rong,Liu Runping,Li Yijie,Li Xiaojiaoyang
Objective: Drug-induced liver injury (DILI) is the leading cause of acute liver failure and poses a significant challenge to human health. Rhubarb ( Rheum officinale Baill . DaHuang) has been clinically used for its heat-clearing and diuresis-promoting effects. However, its toxic effects on different organelles in the liver require further validation. Methods: We analyzed the potential targets affecting hepatotoxicity in rhubarb and the potential damage relationship with five major organelles, including microsomes, mitochondria, endoplasmic reticulum (ER), Golgi apparatus (GA), and lysosomes through Integrated Traditional Chinese Medicine (ITCM)/HERB databases and network pharmacology. We isolated and purified different organelles, incubated them with different fractions and monomers of rhubarb in an adenosine triphosphate (ATP) culture system and examined the structural and functional changes in the organelles using particle size analysis and molecular biological experiments to investigate whether rhubarb affects the damage and rupture of major organelles in the liver. Results: By combining virtual predictions and experimental verification, our research confirmed that emodin isolated from the anthraquinone of rhubarb, catechin in the tannins of rhubarb, and palmitic acid in the organic acids of rhubarb caused the most significant functional and structural damage to the representative organelles. Among all the monomeric compounds, emodin caused the most damage to the microsome, mitochondria, ER, and lysosome; catechin induced microsome and GA damage; and palmitic acid caused the most damage to microsomes and GA in the liver, suggesting that rhubarb components may exert hepatotoxicity through multi-organelle injury. Conclusions: Our findings revealed that rhubarb has varying degrees of damaging effects on different organelles, which in turn affects cellular life activities by impairing organelle morphology and function. This study provides a theoretical basis and technical support for a refined analysis of the toxic components and targets of rhubarb. Export
研究领域:
中药学
·
药理学
·
细胞生物学
·
生物化学
✓ 已验证
DOI
Chinese Herbal Medicines
·
2025
·
IF 8.9
Yijie Li,Shuni Duan,Yinhao Zhang,Runping Liu,Rong Sun,Jianzhi Wu,Zhi Ma,Xiaojiaoyang Li
Objective Drug-induced liver injury (DILI) is an undesirable reaction caused by drugs, herbal medicines or upplements and may lead to acute liver failure. Polygoni Multiflori Radix (PMR, Heshouwu in Chinese) originated from the roots of Polygonum multiflorum is a popular traditional Chinese medicine (TCM) while potential hepatotoxicity limits its clinical application. The present study aims to elucidate the in-depth mechanism of PMR-induced organelle heterogeneity of hepatotoxicity. Methods Network pharmacology and available TCM transcriptomics databases including Integrated Traditional Chinese Medicine (ITCM) and HERB databases were conducted to identify the active ingredients of PMR with the potent ability to injure organelles including microsome, mitochondria, endoplasmic reticulum (ER), Golgi apparatus (GA), and lysosome. Organelles were isolated and cultured with adenosine triphosphate (ATP)-upplemented system. Western blotting and particle size characterization techniques were further performed to clarify the organelle heterogeneity of PMR-induced hepatotoxicity. Results Five representative organelles were isolated from mouse livers or hepatocytes and administrated with PMR-derived active components and monomers. The results of network pharmacology and virtual screening initially identified the components of PMR that may damage different organelles. By combining experimental verification, we found that five organelles studied in this research were the target organelles for flavonoid (FVN)’s affiliated compound quercetin (QC). Mitochondria were damaged mainly by kaempferol, anthraquinone (AQ) and its monomeric components. Meanwhile, QC and emodin showed effective toxicity on endoplasmic reticulum. For microsome, QC remained the most toxic monomer. For Golgi apparatus, trans -stilbene glycosides ( trans- SG), AQ and emodin were the major toxic components in PMR. For lysosomes, total-SG, emodin and QC were the major toxic components in PMR. Conclusion Collectively, our findings revealed the organelle heterogeneity of PMR-induced hepatotoxicity and identified quercetin as a potential toxic component in PMR. This study provides a novel conjunct strategy to screen and discover potential toxic components and shapes the understanding of toxicity warning and clinically safe use of herbal medicines such as PMR.
研究领域:
中药学
·
毒理学
·
肝脏病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ORGANIC & BIOMOLECULAR CHEMISTRY
·
2017
·Vol. 15
·期 27
·5764-5771
·
IF 3.564
Sizhe Zhu,Si Yao,Fengshou Wu,Lijun Jiang,Ka-Leung Wong,Ji Zhou,Kai Wang
Organelle and nucleus dual-targeted anticancer drugs are being increasingly used for efficient cancer therapy as they can attack the double vital sites of tumor cells. In this work, we synthesized and characterized two new porphyrin compounds Pt-Por-RB and Me-Por-RB. The spectral titration results suggest that both Pt-Por-RB and Me-Por-RB bind to DNA efficiently in an intercalation binding mode. Upon irradiation, Pt-Por-RB with low dark-cytotoxicity can rapidly generate singlet oxygen to damage the tumor cells through the process of photodynamic therapy. Compared with Me-Por-RB, Pt-Por-RB was not only internalized in the organelles, but also in the nuclei of HeLa cells, probably due to the presence of platinum complexes, as analyzed using the confocal laser scanning microscope. Thus, with the combination of organelle and nucleus dual-targeting property and high efficiency of singlet oxygen generation, Pt-Por-RB showed a significant therapeutic activity against tumor cells.
研究领域:
药理学
·
肿瘤学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Shujing Wang,Ying Lin,Chengkai Zhang,Tong Zhu,Xiaohe Tian,Dandan Li,Wen Ma,Qiong Zhang,Jieying Wu,Yupeng Tian
In this work, a series of multiphoton terpyridine agents (ZA, ZA-Mex, and ZA-Hex) for fluorescence lifetime imaging microscopy (FLIM) are designed and synthesized. The results from photophysical property research reveal that ZA-Hex, as an N-hexylated terpyridine salt, has stronger three-photon aggregation-induced emission (AIE) properties compared to ZA-Mex due to enhanced intramolecular charge transfer (ICT) performance. All three terpyridine derivatives possess suitable fluorescence intensities and stable fluorescence lifetimes under different pH conditions (pH = 4.0–8.0), thereby performing multiphoton fluorescence lifetime imaging. For biological imaging applications, it is found that ZA shows good lipid droplet (LD) turn-on fluorescence performance, and ZA-Hex could easily accumulate in mitochondria with high specificity. This is the first report of terpyridine salts as three-photon AIE probes used for multiphoton FLIM imaging.
研究领域:
化学
·
生物成像
·
材料科学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2023
·Vol. 11
·期 30
·7134-7143
·
IF 7
Hao-Chi Hao,Gang Zhang,Ru Sun,Yu-Jie Xu,Jian-Feng Ge
Four 1,8-naphthyridine derivatives (1a–1d) with different organelle targeting abilities were obtained using the Knoevenagel condensation reaction of 1,8-naphthyridine with 4-(N,N-diethylamino)benzaldehyde (2a), 4-(N,N-diphenylamino)benzaldehyde (2b), 4-(piperazin-1-yl)benzaldehyde (2c) and 4-(ethyl(4-formylphenyl)amino)-N-(2-((4-methylphenyl)sulfonamido)ethyl)butanamide (2d), respectively. The maximal absorption bands of dyes 1a–1d were observed at 375–447 nm, while their maximum emission peaks were situated at 495–605 nm. The optical properties showed that the fluorescence emission of dyes 1a–1d is shifted toward greater wavelengths as the system polarity (Δf) increased. Meanwhile, with increasing polarity of the mixed 1,4-dioxane/H2O system, the fluorescence intensity of dyes 1a–1d gradually decreased. Furthermore, the fluorescence intensity of 1a–1d enhanced by 12–239 fold as the polarity of 1,4-dioxane/H2O mixtures declined. 1a–1d had a large Stokes shift (up to 229 nm) in polar solvents in comparison to nonpolar solvents. The colocalization imaging experiments demonstrated that dyes 1a–1d (3–10 μM) were located in mitochondria, lipid droplets, lysosomes and the endoplasmic reticulum in living HeLa cells, respectively; and they could monitor the polarity fluctuation of the corresponding organelles. Consequently, this work proposes a molecular design idea with different organelle targeting capabilities based on the same new fluorophore, and this molecular design idea may provide more alternatives for polarity-sensitive fluorescent probes with organelle targeting.
研究领域:
细胞生物学
·
分子生物学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Huihui Zhang,Xiaojiao Zhu,Hong Li,Gang Liu,Junjun Wang,Aidong Wang,Lin Kong,Weiju Zhu,Hongping Zhou
Dynamic tracking of the spatiotemporal coordination among various organelles to in-depth understanding of the mechanism of autophagy have attracted considerable attention. However, the monitor of nucleoli participation in autophagy was somehow neglected. Herein, we report a RNA-targeted bioprobe (ADAP) with high selective permeability into nuclear pore complexes, which induced a two-photon (TP) fluorescence “off-on” response by groove combination with RNA, dynamically monitoring the autophagy process among multiorganelles (nucleoli, mitochondria, and mitochondria-containing lysosomes). This work provides a simple and convenient way to observe the dynamic behavior of multiorganelles during the autophagy process, which benefits the understanding of cellular metabolism.
研究领域:
细胞生物学
·
分子生物学
·
生物成像
✓ 已验证
DOI
PubMed
高影响因子
Nanoscale
·
2017
·Vol. 10
·期 3
·1532-1539
·
IF 7.367
Xiaolong Wu,Liling Ma,Shan Sun,Kai Jiang,Ling Zhang,Yuhui Wang,Haibo Zeng,Hengwei Lin
In this work, a versatile platform for the highly efficient preparation of graphene quantum dots (GQDs) with diverse properties was developed. First of all, an excess amount of oxidants and an additional high temperature step of the Hummers’ method for the synthesis of graphene oxide (GO) was applied to obtain nanosized graphene oxide (NGO). Then, high quality GQDs (quantum yields up to 18.2%) with different photoluminescence emission wavelengths, adjustable hydrophilicity–hydrophobicity, and selective cell organelle imaging capacity can be facilely achieved through a one-pot hydrothermal reaction between the NGO and ammonia, fatty primary amines, or amino-substituted organelle targetable compounds, respectively. The superior features of the as-developed method are extremely high conversion ratio (ca. 60 wt% from graphite to the functional GQDs) and great expandability. Such a high conversion ratio is deemed to be due to effectively decreasing aggregation of the NGO (in comparison with GO) during the post-treatment process. This work provides a robust strategy for the highly efficient preparation of GQDs with diverse properties and functions, and is believed to be beneficial for boosting their applications in the future.
研究领域:
材料科学
·
生物医学
·
化学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2019
·Vol. 43
·期 46
·18251-18258
·
IF 3.069
Pei Yin,Tao Wang,Yuan Yang,Weidong Yin,Shaoxiong Zhang,Zengming Yang,Chunxuan Qi,Hengchang Ma
The chemical structures or positive charge-guided specific cell organelles' staining was an interesting but very challenging research work. In this paper, we synthesized a series of triphenylamine (TPA) derivatives with mono-, di- and tri-pyridinium as substituent groups. By exchanging the negative ions (I−, BF4− and PF6−) between them, a platform containing small compounds was established. After that, mammalian cells of HeLa and plant cells of BY-2 were employed for the study on cell imaging. The results clearly demonstrated that mono-pyridinium derivatives (TPA-Py-I−, TPA-Py-BF4− and TPA-Py-PF6−) are mitochondrion-targeted (for HeLa cells) and cell wall-targeted (for BY-2 cells). However, both di- and tri-pyridinium substituted derivatives (TPA-diPy-BF4− and TPA-triPy-BF4−) were nucleus-targeted for both cell lines. From these important findings, we can conclude that the quantity of positive charge plays a decisive role in different cell organelle-targeted staining. We also declared that our research results were very useful for designing and synthesizing probes with good cellular uptake capability, significantly enhanced targeting ability, and DNA detection.
研究领域:
细胞生物学
·
分子生物学
·
化学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
·
2013
·
IF 3.172
Xiaoxue Song,Huanbin Li,Weijun Tong,Changyou Gao
Encapsulation of pH sensitive fluorophores as reporting molecules provides a powerful approach to visualize the transportation of multilayer capsules. In this study, two pH sensitive dyes (fluorescein and oregon green) and one pH insensitive dye (rhodamine B) were simultaneously labeled on the microcapsules to fabricate ratiometric pH sensors. The fluorescence of the triple-labeled microcapsule sensors was robust and nearly independent of other intracellular species. With a dynamic pH measurement range of 3.3–6.5, the microcapsules can report their localized pH at a real time. Cell culture experiments showed that the microcapsules could be internalized by RAW 246.7 cells naturally and finally accumulated in acidic organelles with a pH value of 5.08 ± 0.59 (mean ± s.d.; n = 162).
研究领域:
生物化学
·
细胞生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
·
2023
·
IF 8.9
Yu-Xi Feng,Peng Tian,Yu-Juan Lin,Dan-Yang Cao,Cheng-Zhi Li,Abid Ullah
The induction of disruption in the electronic transport chain by thiocyanate (SCN − ) leads to an excessive generation of reactive oxygen species (ROS) within rice ( Oryza sativa ). Hydrogen sulfide (H 2 S) assumes a crucial role as a gaseous signaling molecule, holding significant potential in alleviating SCN − -related stress. Nevertheless, there remains a dearth of understanding regarding the intricate interplay between H 2 S and ROS in Oryza sativa amidst SCN − pollution. In this investigation, a hydroponics-based experiment was meticulously devised to explore how H 2 S-mediated modifications influence the genetic feedback network governing ROS metabolism within the subcellular organelles of Oryza sativa when exposed to varying effective concentrations (EC 20 : 24 mg SCN/L; EC 50 : 96 mg SCN/L; EC 75 : 300 mg SCN/L) of SCN − . The findings unveiled the enhanced capacity of Oryza sativa to uptake SCN − under H 2 S + SCN − treatments in comparison to SCN − treatments alone. Notably, the relative growth rate (RGR) of seedlings subjected to H 2 S + SCN − exhibited a superior performance when contrasted with seedlings exposed solely to SCN − . Furthermore, the application of exogenous H 2 S yielded a significant reduction in ROS levels within Oryza sativa tissues during SCN − exposure. To elucidate the intricacies of gene regulation governing ROS metabolism at the mRNA level, the 52 targeted genes were categorized into four distinct types, namely: initial regulatory ROS generation genes (ROS-I), direct ROS scavenging genes (ROS-II), indirect ROS scavenging genes (ROS-III), and lipid oxidation genes (ROS-IV). On the whole, exogenous H 2 S exhibited the capacity to activate the majority of ROS-I ∼ ROS-IV genes within both Oryza sativa tissues at the EC 20 concentration of SCN − . However, genetic positive/negative feedback networks emphasized the pivotal role of ROS-II genes in governing ROS metabolism within Oryza sativa . Notably, these genes were predominantly activated within the cytoplasm, chloroplasts, mitochondria, peroxisomes, and the cell wall.
研究领域:
植物生理学
·
分子生物学
·
环境科学
·
农学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Nano
·
2022
·
IF 18.027
Wei Wei,Huiting Lu,Wenhao Dai,Xiaonan Zheng,Haifeng Dong
Multiplexed profiling of microRNAs’ subcellular expression and distribution is essential to understand their spatiotemporal function information, but it remains a crucial challenge. Herein, we report an encoding approach that leverages combinational fluorescent dye barcodes, organelle targeting elements, and an independent quantification signal, termed Multiplexed Organelles Portrait Barcodes (MOPB), for high-throughput profiling of miRNAs from organelles. The MOPB barcodes consist of heterochromatic fluorescent dye-loaded shell–core mesoporous silica nanoparticles modified with organelle targeting peptides and molecular beacon detection probes. Using mitochondria and endoplasmic reticulum as models, we encoded four Cy3/AMCA ER-MOPB and four Cy5/AMCA Mito-MOPB by varying the Cy3 and Cy5 intensity for distinguishing eight organelles’ miRNAs. Significantly, the MOPB strategy successfully and accurately profiled eight subcellular organelle miRNAs’ alterations in the drug-induced Ca2+ homeostasis breakdown. The approach should allow more widespread application of subcellular miRNAs and multiplexed subcellular protein biomarkers’ monitoring for drug discovery, cellular metabolism, signaling transduction, and gene expression regulation readout.
研究领域:
细胞生物学
·
分子生物学
·
纳米技术
·
药物发现
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）