TY - JOUR
T1 - Spontaneous and multifaceted ATP release from astrocytes at the scale of hundreds of synapses
AU - Hatashita, Yoshiki
AU - Wu, Zhaofa
AU - Fujita, Hirotaka
AU - Kumamoto, Takuma
AU - Livet, Jean
AU - Li, Yulong
AU - Tanifuji, Manabu
AU - Inoue, Takafumi
N1 - Publisher Copyright:
© 2023 Wiley Periodicals LLC.
PY - 2023/9
Y1 - 2023/9
N2 - Astrocytes participate in information processing by releasing neuroactive substances termed gliotransmitters, including ATP. Individual astrocytes come into contact with thousands of synapses with their ramified structure, but the spatiotemporal dynamics of ATP gliotransmission remains unclear, especially in physiological brain tissue. Using a genetically encoded fluorescent sensor, GRABATP1.0, we discovered that extracellular ATP increased locally and transiently in absence of stimuli in neuron–glia co-cultures, cortical slices, and the anesthetized mouse brain. Spontaneous ATP release events were tetrodotoxin-insensitive but suppressed by gliotoxin, fluorocitrate, and typically spread over 50–250 μm2 area at concentrations capable of activating purinergic receptors. Besides, most ATP events did not coincide with Ca2+ transients, and intracellular Ca2+ buffering with BAPTA-AM did not affect ATP event frequency. Clustering analysis revealed that these events followed multiple distinct kinetics, and blockade of exocytosis only decreased a minor group of slow events. Overall, astrocytes spontaneously release ATP through multiple mechanisms, mainly in non-vesicular and Ca2+-independent manners, thus potentially regulating hundreds of synapses all together.
AB - Astrocytes participate in information processing by releasing neuroactive substances termed gliotransmitters, including ATP. Individual astrocytes come into contact with thousands of synapses with their ramified structure, but the spatiotemporal dynamics of ATP gliotransmission remains unclear, especially in physiological brain tissue. Using a genetically encoded fluorescent sensor, GRABATP1.0, we discovered that extracellular ATP increased locally and transiently in absence of stimuli in neuron–glia co-cultures, cortical slices, and the anesthetized mouse brain. Spontaneous ATP release events were tetrodotoxin-insensitive but suppressed by gliotoxin, fluorocitrate, and typically spread over 50–250 μm2 area at concentrations capable of activating purinergic receptors. Besides, most ATP events did not coincide with Ca2+ transients, and intracellular Ca2+ buffering with BAPTA-AM did not affect ATP event frequency. Clustering analysis revealed that these events followed multiple distinct kinetics, and blockade of exocytosis only decreased a minor group of slow events. Overall, astrocytes spontaneously release ATP through multiple mechanisms, mainly in non-vesicular and Ca2+-independent manners, thus potentially regulating hundreds of synapses all together.
KW - astrocytes
KW - calcium imaging
KW - channel-mediated ATP release
KW - exocytosis
KW - extracellular ATP imaging
KW - gliotransmission
KW - neuron–glia interactions
UR - http://www.scopus.com/inward/record.url?scp=85161467658&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85161467658&partnerID=8YFLogxK
U2 - 10.1002/glia.24392
DO - 10.1002/glia.24392
M3 - Article
C2 - 37259810
AN - SCOPUS:85161467658
SN - 0894-1491
VL - 71
SP - 2250
EP - 2265
JO - GLIA
JF - GLIA
IS - 9
ER -