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Regulated ATP release from astrocytes through lysosome exocytosis

Nature Cell Biology volume 9, pages 945–953 (2007)Cite this article

Abstract

Release of ATP from astrocytes is required for Ca2+ wave propagation among astrocytes1,2,3 and for feedback modulation of synaptic functions2,4,5. However, the mechanism of ATP release and the source of ATP in astrocytes are still not known. Here we show that incubation of astrocytes with FM dyes leads to selective labelling of lysosomes. Time-lapse confocal imaging of FM dye-labelled fluorescent puncta, together with extracellular quenching and total-internal-reflection fluorescence microscopy (TIRFM), demonstrated directly that extracellular ATP or glutamate induced partial exocytosis of lysosomes, whereas an ischaemic insult with potassium cyanide induced both partial and full exocytosis of these organelles. We found that lysosomes contain abundant ATP, which could be released in a stimulus-dependent manner. Selective lysis of lysosomes abolished both ATP release and Ca2+ wave propagation among astrocytes, implicating physiological and pathological functions of regulated lysosome exocytosis in these cells.

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Figure 1: Confocal images showing that prolonged loading of cultured astrocytes with FM dyes specifically labels lysosomes.
Figure 2: Destaining of the FM dye-labelled puncta in astrocytes by ATP and KCN.
Figure 3: Ca2+ dependence and TIRFM imaging of lysosome exocytosis in astrocytes.
Figure 4: Lysosomes in astrocytes contain abundant ATP.
Figure 5: Time course of the release of ATP, β-hexosaminidase (hexosam.) and lactate dehydrogenase (LDH) from cultured astrocytes and hippocampal slices.

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Acknowledgements

We thank M-m. Poo for critical comments on the manuscript, and Z. Zhou, X. K. Chen and Y. Zhou for valuable discussion. This work was supported by grants from the Major State Basic Research Program of China (G200077800 and 2006CB806600) and the National Natural Science Foundation of China (30321002).

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Author notes

  1. Zhijun Zhang, Gang Chen and Wei Zhou: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Neuroscience and Key Laboratory of Neurobiology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China

    Zhijun Zhang, Gang Chen, Wei Zhou, Aihong Song & Shumin Duan

  2. Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, 226001, Jiangsu, China

    Gang Chen & Xiao-song Gu

  3. Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science & Technology, Wuhan, 430074, China

    Wei Zhou & Qingming Luo

  4. National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China

    Tao Xu

  5. Department of Neurology, Tongji Hospital, Tongji Medical Scool, Huazhong University of Science and Technology, Wuhan, 430030, China

    Wei Wang

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Zhang, Z., Chen, G., Zhou, W. et al. Regulated ATP release from astrocytes through lysosome exocytosis. Nat Cell Biol 9, 945–953 (2007). https://doi.org/10.1038/ncb1620

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