Vesicular nucleotide transporter is involved in ATP storage of secretory lysosomes in astrocytes

Manami Oya, Tetsuya Kitaguchi, Yu Yanagihara, Rika Numano, Masaki Kakeyama, Kazuya Ikematsu, Takashi Tsuboi

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Recent studies have suggested that astrocytes release gliotransmitters (i.e., ATP, L-glutamate, D-serine, and peptide hormones) and participate actively in synaptic functioning. Although ATP release from astrocytes modulates the activity of neurons, the mechanisms regulating the ATP release from astrocytes and the source of ATP in astrocytes are not well understood. Recently a vesicular nucleotide transporter (VNUT)/solute carrier family 17, member 9 (SLC17A9) has been identified as a mediator of the active accumulation of ATP into vesicles. Here we show by immunocytochemical analysis under confocal microscope and live cell imaging under total internal reflection fluorescence microscope that lysosome-associated VNUT is responsible for ATP release in astrocytes. VNUT was expressed in both primary cultured cortical astrocytes and glioma cell line C6 cells, and mainly localized on lysosome in the cells. We found that VNUT-associated secretory lysosomes do not fully collapse into the plasma membrane after lysosomal exocytosis. We also found that inhibition of VNUT function by Evans Blue decreased ATP uptake into secretory lysosomes. Depletion and inhibition of endogenous VNUT by small interference RNA and Evans Blue, respectively decreased the amount of ATP release from the cells, whereas overexpression of VNUT increased it. Taken together, these findings indicate that the participation of VNUT in ATP storage in secretory lysosomes during lysosomal exocytosis of ATP from astrocytes.

Original languageEnglish
Pages (from-to)145-151
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume438
Issue number1
DOIs
Publication statusPublished - 2013 Aug 16
Externally publishedYes

Fingerprint

Lysosomes
Astrocytes
Nucleotides
Adenosine Triphosphate
Evans Blue
Exocytosis
Microscopes
Peptide Hormones
Cell membranes
RNA Interference
Glioma
Serine
Neurons
Glutamic Acid
Fluorescence
Cells
Cell Membrane
RNA
Imaging techniques
Cell Line

Keywords

  • Astrocytes
  • ATP
  • Exocytosis
  • Lysosome
  • Vesicular nucleotide transporter

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Vesicular nucleotide transporter is involved in ATP storage of secretory lysosomes in astrocytes. / Oya, Manami; Kitaguchi, Tetsuya; Yanagihara, Yu; Numano, Rika; Kakeyama, Masaki; Ikematsu, Kazuya; Tsuboi, Takashi.

In: Biochemical and Biophysical Research Communications, Vol. 438, No. 1, 16.08.2013, p. 145-151.

Research output: Contribution to journalArticle

Oya, Manami ; Kitaguchi, Tetsuya ; Yanagihara, Yu ; Numano, Rika ; Kakeyama, Masaki ; Ikematsu, Kazuya ; Tsuboi, Takashi. / Vesicular nucleotide transporter is involved in ATP storage of secretory lysosomes in astrocytes. In: Biochemical and Biophysical Research Communications. 2013 ; Vol. 438, No. 1. pp. 145-151.
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