Scientific Reports
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2015
·Vol. 5
·期 1
·1-16
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IF 5.578
Wan Min,Zhang Wenhua,Tian Yangli,Xu Chanjuan,Xu Tao,Liu Jianfeng,Zhang Rongying
Endocytosis and postendocytic sorting of G-protein-coupled receptors (GPCRs) is important for the regulation of both their cell surface density and signaling profile. Unlike the mechanisms of clathrin-dependent endocytosis (CDE), the mechanisms underlying the control of GPCR signaling by clathrin-independent endocytosis (CIE) remain largely unknown. Among the muscarinic acetylcholine receptors (mAChRs), the M4 mAChR undergoes CDE and recycling, whereas the M2 mAChR is internalized through CIE and targeted to lysosomes. Here we investigated the endocytosis and postendocytic trafficking of M2 mAChR based on a comparative analysis of the third cytoplasmic domain in M2 and M4 mAChRs. For the first time, we identified that the sequence 374 KKKPPPS 380 servers as a sorting signal for the clathrin-independent internalization of M2 mAChR. Switching 374 KKKPPPS 380 to the i3 loop of the M4 mAChR shifted the receptor into lysosomes through the CIE pathway; and therefore away from CDE and recycling. We also found another previously unidentified sequence that guides CDE of the M2 mAChR, 361 VARKIVKMTKQPA 373 , which is normally masked in the presence of the downstream sequence 374 KKKPPPS 380 . Taken together, our data indicate that endocytosis and postendocytic sorting of GPCRs that undergo CIE could be sequence-dependent.
研究领域:
细胞生物学
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分子生物学
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神经科学
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DOI
PubMed
高影响因子
Advanced Healthcare Materials
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2014
·Vol. 4
·期 3
·413-419
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IF 4.88
Ye Tian,Man Wu,Xiangxiang Liu,Zhi Liu,Quan Zhou,Zhongwei Niu,Yong Huang
Viral nanoparticles have attracted extensive research interests in diverse applications of diagnosis and therapy. In particular, filamentous M13 bacteriophages have shown great potential in biomedical applications. However, its pathways entering into cells still remain unclear, and this greatly hinders its further use as a drug or gene carrier. Here, a ratiometric M13 pH probe is designed by conjugating two fluorescent dyes onto the surface of M13. Since the intensity ratio is not influenced by probe concentration, ion strength, temperature, photobleaching, and optical path length, this ratiometric probe can be used to investigate the intracellular pH map of M13. More importantly, the internalization mechanism of M13 can be elucidated. It is found that this filamentous phage shows great cell-type dependence in interaction with cells and internalization mechanism. The phage tends to be bounded on the cell membrane of only epithelial cells, not endothelial cells. Furthermore, the M13 phage enters into cells through endocytosis with specific mechanism: clathrin-mediated endocytosis and macropinocytosis for HeLa; vesicular transport, clathrin-mediated endocytosis, and macropinocytosis for MCF-7; caveolae-mediated endocytosis for human dermal microvascular endothelial cell (HDMEC). This work provides key notes for cancer diagnosis and therapy based on filamentous bacteriophage, especially for design of pH-sensitive drug delivery systems.
研究领域:
生物技术
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纳米技术
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细胞生物学
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药学
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DOI
PubMed
高影响因子
Nanomedicine
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2019
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IF 4.717
Xiao Wei,Rui Wei,Guiying Jiang,Yijuan Jia,Hua Lou,Zongyuan Yang,Danfeng Luo,Quanfu Huang,Sen Xu,Xin Yang,Ying Zhou,Xiaoting Li,Teng Ji,Junbo Hu,Ling Xi,Ding Ma,Fei Ye,Qinglei Gao
Aim: To investigate the influence of tissue mechanics on the cellular uptake efficiency of nanoparticles (NPs) in cancer. Materials & methods: Collagen-coated polyacrylamide gels were prepared as model substrates. Coumarin 6-loaded poly(lactic-co-glycolic) acid micelles (C6-NPs) were prepared to investigate the cellular uptake of NPs. Results: We demonstrated that substrate stiffness modulated the cellular uptake of NPs of cancer. Mechanistically, mechanical cues exerted influence on the clathrin-mediated endocytosis and caveolae-mediated endocytosis pathways, which mediated stiffness-regulated cellular uptake of NPs. Conclusion: Our findings shed light on the regulatory role of the mechanical cues on the cellular uptake of NPs and will facilitate the selection of clinical patients who might benefit from a given nanotherapy.
研究领域:
生物医学工程
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癌症研究
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纳米技术
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细胞生物学
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DOI
PubMed
高影响因子
RSC Advances
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2016
·Vol. 6
·期 99
·96635-96643
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IF 3.289
Peng Zhao,Meng Cao,Lina Song,Hao Wu,Ke Hu,Bo Chen,Qiwei Wang,Ning Gu
Magnetic nanoparticles (MNPs) are widely used in biomedical applications in vivo. However, the endocytosis of mononuclear phagocyte system (MPS) is still a major challenge for MNPs delivery in vivo. MIM (MTSS1) is a multifunctional scaffold protein to regulate both actin dynamics and membrane dynamics, which may decide the readily particles clear ability of MPS and be implicated in the cellular endocytosis. To find out the exact role of MIM plays in the nanoparticle uptake process of macrophages, we established a MIM knock-down cell line RAW 264.7MIM-. The endocytosis rate and efficiency were detected to find out the differences between the normal RAW 264.7 and RAW 264.7MIM- cell after 24 h of exposure to the Fe2O3@DMSA MNPs (70 nm hydrodynamic size). The results indicated that the clathrin-mediated endocytosis of RAW 264.7MIM- cell involves fewer particles than normal RAW 264.7 cells with significant differences in the concentration ranging from 100 to 200 μg mL−1. Therefore, knock-down the MIM expression in macrophage would affect the endocytosis process to iron oxide nanoparticles mainly in clathrin-mediated pathway. As a whole, our results presented here illustrated that MIM plays a positive role in the cellular endocytosis process of MNPs, which is a meaningful molecular basis for biomedical applications of nanomaterials.
研究领域:
生物医学工程
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细胞生物学
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纳米技术
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产品关联
DOI
高影响因子
ENVIRONMENTAL POLLUTION
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2021
·
IF 8.071
Qing Yang,Yongqiang Liu,Lihong Wang,Qing Zhou,Mengzhu Cheng,Jiahong Zhou,Xiaohua Huang
Aggravating the pollution of microcystins (MCs) in freshwater environments is detrimental to aquatic living organisms and humans, and thus threatens the stability of ecosystems. Some environmental factors have been verified to promote the production of MCs in Microcystis aeruginosa , thereby aggravating the pollution of MCs. However, the effects of cerium (Ce), the most abundant rare earth element in global water environments, on the production of MCs in M. aeruginosa are unknown. Here, Lake Taihu water was selected as a representative of freshwater environments. By using interdisciplinary methods, it was found that: (1) the exposure level of Ce [Ce(III) and Ce(IV)] in Lake Taihu water is in the range of 0.271–0.282 μg/L; (2) Ce exposure in Lake Taihu water promoted the contents of three main MCs (MC-LR, MC-LW and MC-YR) in M. aeruginosa and water; (3) a cellular mechanism of Ce promoting the production of MCs in M. aeruginosa in Lake Taihu water was suggested: Ce enhanced endocytosis in cells of M. aeruginosa to promote the essential element uptake by M. aeruginosa for MC synthesis. Thus, Ce exposure in Lake Taihu water aggravates the pollution of MCs via enhancing endocytosis in cells of M. aeruginosa . The results provide reference for assessing the environmental risk of Ce in water environments, investigating the mechanism of the pollution of MCs induced by environmental factors, and developing strategies aimed at preventing and controlling the pollution of MCs.
研究领域:
环境科学
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生物学
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产品关联
DOI
PubMed
高影响因子
Advanced Healthcare Materials
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2025
·
IF 9.6
Weibo Zhang,Chong Chen,Ran Wang,Yixuan Li,Yinhua Zhu,Jiazeng Sun,Yongting Luo,Junjie Luo,Liang Zhao,Yanan Sun,Huiyuan Guo,Hao Zhang,Bing Fang,Yao Hu,Rong Liu,Xiaoyu Wang,Jiayue Guo,Ping Liu,Peng An,Pengjie Wang,Juan Chen,Fazheng Ren
Human serum albumin (HSA) plays an indispensable role as a carrier of endogenous and exogenous substances in vivo. The efficiency of endocytosis determines the transport capacity. However, the potential for enhancing the efficiency of HSA endocytosis remains largely unknown. This research represented a substantial advancement, revealing that Fe 2+ markedly increased HSA endocytosis. Moreover, Fe 2+ facilitated the endocytosis and transport of HSA in vivo. The enhancement in HSA uptake facilitated by Fe 2+ is markedly decreased following iron chelation, indicating a specific interaction between Fe 2 ⁺ and HSA that promoted endocytosis. Moreover, the study elucidated that FcRn and caveolin regulated HSA endocytosis under normal conditions. Yet, in the presence of Fe 2 ⁺, endocytosis shifted toward clathrin- and caveolin-enriched membrane domains, requiring both divalent metal transporter 1 (DMT1) and FcRn for efficient process completion. Thereby a new pathway is discovered for Fe 2+ -dependent HSA endocytosis involving DMT1 and clathrin. This findings highlighted the crucial role of iron in enhancing HSA endocytosis and transport, establishing Fe 2+ as a vital endocytic enhancer that required binding to HSA to substantially improve its uptake. This insight into the mechanism of HSA endocytosis not only expands the understanding of protein transport but also opens new avenues for therapeutic intervention.
研究领域:
生物化学
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细胞生物学
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医学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2022
·
IF 8.025
Weibo Zhang,Shucheng Zhang,Chong Chen,Ning Liu,Dong Yang,Pengjie Wang,Fazheng Ren
Pea albumin (PA) can reach the intestine in the active form because it is highly resistant to gastric acid and proteolytic enzymes after their oral intake, which can supply various bioactivities. However, there is no detailed knowledge of the intestinal cell uptake about PA. The aim of this work was to study the internalization mechanism and intracellular trafficking route of PA. The uptake of PA-cyanine 5.5 NHS ester (Cy5.5) was a time-dependent and concentration-dependent process in Caco-2 cells. Endocytosis inhibitors or small interfering RNA (siRNA) techniques revealed that the internalization of PA-Cy5.5 was energy-dependent and mediated by caveolin-mediated endocytosis. Furthermore, we observed colocalization of PA-Cy5.5 and its subcellular localization in Caco-2 cells by using confocal laser scanning microscopy, which revealed that the intracellular trafficking process of PA-Cy5.5 was related to endoplasmic reticulum, Golgi, and lysosome. Interestingly, PA can alleviate lipopolysaccharide -induced ER stress, which may be the main reason why pea albumin is anti-inflammatory. Overall, our findings suggest caveolin may be critical for PA uptake in enterocytes and could contribute to explore the bioactivities mechanism of pea albumin in body.
研究领域:
营养学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2020
·
IF 9.038
Yingxin Chen,Yun Ling,Xinyu Li,Jiani Hu,Chengjin Cao,Defu He
Microplastics (MPs) are persistent contaminants in aquatic environments. Microalgae, as the main phytoplankton and primary producers, usually co-exist with MPs. Despite previous studies that have proved the interaction of MPs and microalgae, it is largely unknown whether MPs can be uptake into cells of microalgae. In this study, both marine P. helgolandica var. tsingtaoensis and freshwater microalgae S. quadricauda were respectively exposed to 10 mg/L polystyrene microbeads with five diameter sizes: 1.0, 2.0, 3.0, 4.0, and 5.0 μm. Confocal laser scanning and 3D image analysis showed that mean 24.0 % or 11.3 % cells of P. helgolandica var. tsingtaoensis contained 1.0 μm or 2.0 μm MPs after 72 h exposure. While mean 43.3 % or 15.3 % of S. quadricauda individuals engulfed 1.0 μm or 2.0 μm MPs within cells. But, none of 3.0–5.0 μm MPs were observed within algal cells. These results demonstrate the size-dependent cellular internalization of MPs in microalgae. Exposure to 1.0–2.0 μm PS MPs caused a significant reduction in the density of microalgae and influenced photosynthesis, which suggests cellular internalization of MPs can influence algal fertility and growth. This discovery first confirms cellular internalization of MPs in phytoplankton, of significance for the fate and eco-toxicity of MPs in the aquatic ecosystem.
研究领域:
环境科学
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生物学
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DOI
PubMed
高影响因子
FASEB JOURNAL
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2020
·Vol. 34
·期 3
·4653-4669
·
IF 4.966
Jian Wang,Yangyang Li,Shouyu Wang,Fei Liu
Transmissible gastroenteritis virus (TGEV) is a swine enteropathogenic coronavirus that causes significant economic losses in swine industry. Current studies on TGEV internalization mainly focus on viral receptors, but the internalization mechanism is still unclear. In this study, we used single-virus tracking to obtain the detailed insights into the dynamic events of the TGEV internalization and depict the whole sequential process. We observed that TGEVs could be internalized through clathrin- and caveolae-mediated endocytosis, and the internalization of TGEVs was almost completed within ~2 minutes after TGEVs attached to the cell membrane. Furthermore, the interactions of TGEVs with actin and dynamin 2 in real time during the TGEV internalization were visualized. To our knowledge, this is the first report that single-virus tracking technique is used to visualize the entire dynamic process of the TGEV internalization: before the TGEV internalization, with the assistance of actin, clathrin, and caveolin 1 would gather around the virus to form the vesicle containing the TGEV, and after ~60 seconds, dynamin 2 would be recruited to promote membrane fission. These results demonstrate that TGEVs enter ST cells via clathrin- and caveolae-mediated endocytic, actin-dependent, and dynamin 2-dependent pathways.
研究领域:
病毒学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2018
·
IF 3.909
Tsogzolmaa Ganbold,Shuqin Han,Agula Hasi,Huricha Baigude
Natural carbohydrate polymer-based nanoparticles have great biocompatibility that is required for the safe delivery of various drugs including nucleic acid therapeutics. Herein, we designed curdlan-based nanoparticles for cancer cell targeted delivery of short interfering RNA (siRNA). iRGD peptide conjugated 6-amino-6-deoxy curdlan specifically delivered siRNA to integrin expressing cancer cells. Incubation of cancer cells with free iRGD peptide competitively blocked cellular uptake of the iRGD functionalized curdlan nanoparticles. Chloroquine but not nystatin inhibited cellular uptake of iRGD functionalized curdlan nanoparticles, indicating that the iRGD peptide conjugated curdlan nanoparticles were internalized through the receptor (clathrin)-mediated endocytosis. Moreover, a disease related gene Plk1 was substantially knocked down by siRNA carried by 6AC-iRGD nanoparticles in HepG2 cells. Our data suggested that iRGD functionalized curdlan may provide a biocompatible carrier for siRNA delivery.
研究领域:
生物医学工程
·
分子生物学
·
药剂学
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DOI
PubMed
高影响因子
NanoImpact
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2025
·
IF 5.5
Yuan-Yuan Liu,Jie Liu,Yuan Guo,Qiangqiang Zhang,Aoneng Cao,Haifang Wang
Nanoplastics enter the human body mainly by ingestion through the gastrointestinal tract and thus the uptake and release of nanoplastics in intestinal cells have been studied. However, the fate of nanoplastics in intestinal cells remains poorly understood, particularly how they are exocytosed. Herein, we investigated the uptake, distribution, and exocytosis of nanoplastics in Caco-2 cells using 70 nm red fluorescent polystyrene (R70PS) as a nanoplastic model. The results show that R70PS readily enters Caco-2 cells and the content per cell peaks at around 24 h, but the total intracellular content in all cells increases continuously over 72 h. In addition, the uptake mechanisms change over incubation time, i.e. R70PS entered Caco-2 cells via both the energy-independent pathway and the energy-dependent caveolae-mediated endocytosis and macropinocytosis at 4 h incubation, but almost all R70PS entered cells in an energy-dependent manner via caveolae-mediated endocytosis, macropinocytosis, and clathrin-mediated endocytosis at 12 h incubation. Most of the intracellular R70PS accumulated in lysosomes, but R70PS also entered the mitochondria and its level increased over time. Approximately 45 % of the intracellular R70PS could be cleared from the cells within 12 h, mainly via the lysosomal pathway. Exocytosis was also associated with autophagy and was facilitated by the increase in the number of mitochondria and lysosomes, but inhibited by serum in the medium. Our findings deepen the understanding of the interaction between nanoplastics and intestinal cells, which is helpful for the risk assessment of nanoplastics.
研究领域:
生物学
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医学
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毒理学
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产品关联
DOI
PubMed
高影响因子
Food Bioscience
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2025
·
IF 5.9
Long Cheng,Zhixin Xie,Ming Cao,Shuaifan Qin,Yongshi Chen,Bo Tian,Zhibiao Feng
As a new type of nanomaterial, carbon dots (CDs) have demonstrated potential in the field of biological imaging. Currently, research utilizing them for labeling microorganisms is confined to simple imaging, with limited studies exploring their application in detecting cell viability and organelle targeting. Yeast, serving as both a fermenting agent and a model cell, constitutes an ideal research subject This study synthesized ascorbic acid-based CDs (A-CDs) using citric acid, ascorbic acid, and ethylenediamine as raw materials via hydrothermal method. A-CDs were uniformly dispersed, with an average particle size of 3.28 nm. The surface functional groups, such as NH 2 and -OH, endowed them with hydrophilicity. Additionally, A-CDs exhibited concentration-dependence, good photostability, and biocompatibility. By labeling yeast with A-CDs, it was observed that cells in dead, alive, and intermediate states exhibited different imaging effects, which were highly correlated with the concentration of A-CDs entering the cell due to variations in cell membrane permeability. Through co-localization experiments with A-CDs and DAPI, it was confirmed that A-CDs could target and label cell nucleus. Preliminary endocytosis experiments indicated that A-CDs primarily entered cells via clathrin-mediated and lipid raft-mediated endocytosis. The ability of A-CDs to discriminate between cell viability and label nuclei offers new perspectives on the application of CDs in the field of biological imaging.
研究领域:
生物成像
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纳米材料科学
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微生物学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）