Constant Conversion Rate of Endolysosomes Revealed by a pH-Sensitive Fluorescent Probe.

Endolysosome dynamics plays an important role in autophagosome biogenesis. Hence, imaging the subcellular dynamics of endolysosomes using high-resolution fluorescent imaging techniques would deepen our understanding of autophagy and benefit the development of pharmaceuticals against endosome-related diseases. Taking advantage of the intramolecular charge-transfer mechanism, herein we report a cationic quinolinium-based fluorescent probe (PyQPMe) that exhibits excellent pH-sensitive fluorescence in endolysosomes at different stages of interest. A systematic photophysical and computational study on PyQPMe was carried out to rationalize its highly pH-dependent absorption and emission spectra. The large Stokes shift and strong fluorescence intensity of PyQPMe can effectively reduce the background noise caused by excitation light and microenvironments and provide a high signal-to-noise ratio for high-resolution imaging of endolysosomes. By applying PyQPMe as a small molecular probe in live cells, we were able to reveal a constant conversion rate from early endosomes to late endosomes/lysosomes during autophagy at the submicron level.

[1]  Yuncong Chen,et al.  Super-Resolution Imaging of Mitochondrial HClO during Cell Ferroptosis Using a Near-Infrared Fluorescent Probe. , 2022, Analytical chemistry.

[2]  J. Guan,et al.  Light-activated mitochondrial fission through optogenetic control of mitochondria-lysosome contacts , 2022, Nature Communications.

[3]  C. K. Nandi,et al.  Ultralong-Term Super-Resolution Tracking of Lysosomes in Brain Organoids by Near-Infrared Noble Metal Nanoclusters , 2022, ACS Materials Letters.

[4]  Chunya Li,et al.  Near-Infrared in and out: Observation of Autophagy during Stroke via a Lysosome-Targeting Two-Photon Viscosity-Dependent Probe. , 2022, Analytical chemistry.

[5]  J. Diao,et al.  Super‐resolution analyzing spatial organization of lysosomes with an organic fluorescent probe , 2022, bioRxiv.

[6]  Kangqiang Qiu,et al.  De Novo Design of A Membrane‐Anchored Probe for Multidimensional Quantification of Endocytic Dynamics , 2022, Advanced healthcare materials.

[7]  T. Takebe,et al.  Simultaneous Zn2+ tracking in multiple organelles using super-resolution morphology-correlated organelle identification in living cells , 2021, Nature Communications.

[8]  Zhaohui Li,et al.  One Stone, Three Birds: pH Triggered Transformation of Aminopyronine and Iminopyronine Based Lysosome Targeting Viscosity Probe for Cancer Visualization. , 2020, Analytical chemistry.

[9]  Zijian Guo,et al.  A dual-labeling probe to track functional mitochondria–lysosome interactions in live cells , 2020, Nature Communications.

[10]  H. Nakatogawa.  Mechanisms governing autophagosome biogenesis , 2020, Nature Reviews Molecular Cell Biology.

[11]  I. González-Domínguez,et al.  Quantification of the HIV‐1 virus‐like particle production process by super‐resolution imaging: From VLP budding to nanoparticle analysis , 2020, Biotechnology and bioengineering.

[12]  Qing Zhu,et al.  A dicyanoisophorone-based, near-infrared, lysosome-targeting pH sensor with an extremely large Stokes shift. , 2020, Analytical biochemistry.

[13]  R. Pfuetzner,et al.  Prefused lysosomes cluster on autophagosomes regulated by VAMP8 , 2020, Cell Death & Disease.

[14]  Qixin Chen,et al.  Nanoscale monitoring of mitochondria and lysosome interactions for drug screening and discovery , 2019, Nano Research.

[15]  C. Dong,et al.  A lysosome-targeting and polarity-specific fluorescent probe for cancer diagnosis. , 2019, Chemical communications.

[16]  J. Guan,et al.  Quantitative Analysis of Interactive Behavior of Mitochondria and Lysosomes Using Structured Illumination Microscopy , 2018, bioRxiv.

[17]  Xingdong Zhou,et al.  Chloroquine inhibits autophagic flux by decreasing autophagosome-lysosome fusion , 2018, Autophagy.

[18]  G. van den Bogaart,et al.  Endosomal and Phagosomal SNAREs. , 2018, Physiological reviews.

[19]  C. Eggeling,et al.  Super-resolution fluorescence microscopy studies of human immunodeficiency virus , 2018, Retrovirology.

[20]  Saran Long,et al.  A ratiometric fluorescence probe for lysosomal polarity. , 2018, Biomaterials.

[21]  Meng Zhang,et al.  Cell-permeable organic fluorescent probes for live-cell long-term super-resolution imaging reveal lysosome-mitochondrion interactions , 2017, Nature Communications.

[22]  Hui Chao,et al.  Organelle-targeting metal complexes: From molecular design to bio-applications , 2017, Coordination Chemistry Reviews.

[23]  Xiaohe Tian,et al.  A two-photon fluorescent probe for real-time monitoring of autophagy by ultrasensitive detection of the change in lysosomal polarity. , 2017, Chemical communications.

[24]  Young‐Tae Chang,et al.  Development of pH-Responsive BODIPY Probes for Staining Late Endosome in Live Cells. , 2016, ChemPlusChem.

[25]  Young‐Tae Chang,et al.  Discerning the Chemistry in Individual Organelles with Small-Molecule Fluorescent Probes. , 2016, Angewandte Chemie.

[26]  Kangqiang Qiu,et al.  Biscylometalated iridium(iii) complexes target mitochondria or lysosomes by regulating the lipophilicity of the main ligands. , 2016, Dalton transactions.

[27]  S. Ryu,et al.  Optogenetically enhanced pituitary corticotroph cell activity post-stress onset causes rapid organizing effects on behaviour , 2016, Nature Communications.

[28]  K. Y. Zhang,et al.  A Mitochondria-Targeted Photosensitizer Showing Improved Photodynamic Therapy Effects Under Hypoxia. , 2016, Angewandte Chemie.

[29]  Mike Heilemann,et al.  Super‐resolved insights into human immunodeficiency virus biology , 2016, FEBS letters.

[30]  W. Gallagher,et al.  Lysosome triggered near-infrared fluorescence imaging of cellular trafficking processes in real time , 2016, Nature Communications.

[31]  R. Borlinghaus,et al.  HyVolution—the smart path to confocal super-resolution , 2016, Nature Methods.

[32]  Eric Bakker,et al.  Selective Imaging of Late Endosomes with a pH-Sensitive Diazaoxatriangulene Fluorescent Probe. , 2016, Journal of the American Chemical Society.

[33]  S. Mitani,et al.  Negative regulation of phosphatidylinositol 3-phosphate levels in early-to-late endosome conversion , 2016, The Journal of cell biology.

[34]  Kangqiang Qiu,et al.  Mitochondria-specific imaging and tracking in living cells with two-photon phosphorescent iridium(iii) complexes. , 2015, Journal of materials chemistry. B.

[35]  Jim A. Thomas.  Optical imaging probes for biomolecules: an introductory perspective. , 2015, Chemical Society reviews.

[36]  Tao Zhang,et al.  A ratiometric lysosomal pH probe based on the naphthalimide-rhodamine system. , 2015, Journal of materials chemistry. B.

[37]  Junying Yuan,et al.  Pharmacologic agents targeting autophagy. , 2015, The Journal of clinical investigation.

[38]  Suming Chen,et al.  Lysosomal pH rise during heat shock monitored by a lysosome-targeting near-infrared ratiometric fluorescent probe. , 2014, Angewandte Chemie.

[39]  A. Tiwari,et al.  pH-activatable near-infrared fluorescent probes for detection of lysosomal pH inside living cells. , 2014, Journal of materials chemistry. B.

[40]  Frederick R Maxfield,et al.  Role of endosomes and lysosomes in human disease. , 2014, Cold Spring Harbor perspectives in biology.

[41]  James H. Hurley,et al.  Atomistic Autophagy: The Structures of Cellular Self-Digestion , 2014, Cell.

[42]  Jun-Ying Miao,et al.  A new fluorescent pH probe for imaging lysosomes in living cells. , 2014, Bioorganic & medicinal chemistry letters.

[43]  G. Milligan,et al.  Predicting small molecule fluorescent probe localization in living cells using QSAR modeling. 1. Overview and models for probes of structure, properties and function in single cells , 2013, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[44]  W. Yang,et al.  Spatiotemporally controlled induction of autophagy-mediated lysosome turnover , 2013, Nature Communications.

[45]  Junying Yuan,et al.  Live imaging and single-cell analysis reveal differential dynamics of autophagy and apoptosis , 2013, Autophagy.

[46]  Yi Xiao,et al.  Activatable rotor for quantifying lysosomal viscosity in living cells. , 2013, Journal of the American Chemical Society.

[47]  Masaaki Komatsu,et al.  Autophagy: Renovation of Cells and Tissues , 2011, Cell.

[48]  Noel M. O'Boyle,et al.  cclib: A library for package‐independent computational chemistry algorithms , 2008, J. Comput. Chem..

[49]  D. Klionsky,et al.  Autophagy: molecular machinery for self-eating , 2005, Cell Death and Differentiation.

[50]  R. Nixon.  Endosome function and dysfunction in Alzheimer's disease and other neurodegenerative diseases , 2005, Neurobiology of Aging.

[51]  S. Emr,et al.  Autophagy as a regulated pathway of cellular degradation. , 2000, Science.

[52]  V. Marjomäki,et al.  At reduced temperature, endocytic membrane traffic is blocked in multivesicular carrier endosomes in rat cardiac myocytes. , 1998, European journal of cell biology.

[53]  T. Kirchhausen,et al.  Wortmannin alters the transferrin receptor endocytic pathway in vivo and in vitro. , 1996, Molecular biology of the cell.

[54]  S. Corvera,et al.  Potential sites of PI-3 kinase function in the endocytic pathway revealed by the PI-3 kinase inhibitor, wortmannin , 1996, The Journal of cell biology.

[55]  K Schulten,et al.  VMD: visual molecular dynamics. , 1996, Journal of molecular graphics.

[56]  Axel D. Becke,et al.  Density-functional thermochemistry. I. The effect of the exchange-only gradient correction , 1992 .

[57]  P. D. Hart,et al.  Ammonium chloride, an inhibitor of phagosome-lysosome fusion in macrophages, concurrently induces phagosome-endosome fusion, and opens a novel pathway: studies of a pathogenic mycobacterium and a nonpathogenic yeast , 1991, The Journal of experimental medicine.

[58]  A. Becke,et al.  Density-functional exchange-energy approximation with correct asymptotic behavior. , 1988, Physical review. A, General physics.

[59]  Parr,et al.  Development of the Colle-Salvetti correlation-energy formula into a functional of the electron density. , 1988, Physical review. B, Condensed matter.

[60]  J. Markham,et al.  Studies on Pyridines: I. The Basicity of Pyridine Bases , 1951 .

[61]  Juyoung Yoon,et al.  A review: the trend of progress about pH probes in cell application in recent years. , 2016, The Analyst.

[62]  Haoxing Xu,et al.  Lysosomal physiology. , 2015, Annual review of physiology.

[63]  Sergio Grinstein,et al.  Sensors and regulators of intracellular pH , 2010, Nature Reviews Molecular Cell Biology.

[64]  C. Duve,et al.  Functions of lysosomes. , 1966, Annual review of physiology.