Optical detection of synaptically induced glutamate transport in hippocampal slices

Satoshi Kojima, Takeshi Nakamura, Takahisa Nidaira, Kyoko Nakamura, Noriko Ooashi, Etsuro Ito, Kei Watase, Kohichi Tanaka, Keiji Wada, Yoshihisa Kudo, Hiroyoshi Miyakawa

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84 Citations (Scopus)

Abstract

Although it has long been believed that glial cells play a major role in transmitter uptake at synapses in the CNS, the relative contribution of glial and neuronal cells to reuptake of synaptically released glutamate has been unclear. Recent identification of the diverse glutamate transporter subtypes provides an opportunity to examine this issue. To monitor glutamate transporter activity, we optically detected synaptically induced changes of membrane potential from hippocampal CA1 field in slice preparations using a voltage-sensitive dye, RH155. In the presence of ionotropic glutamate- receptor blockers, synaptic inputs gave rise to a slow depolarizing response (SDR) in the dendritic field. The amplitude of SDR correlated well with presynaptic activities, suggesting that it was related to transmitter release. The SDR was found to be caused by the activities of glutamate transporters because it was not affected by blockers for GABA(A), nACh, 5- HT3, P(2X), or metabotropic glutamate receptors but was greatly reduced by dihydrokainate (DHK), a specific blocker for GLT-1 transporter, and by D,L- threo-β-hydroxyaspartate (THA), a blocker for EAAC, GLAST, and GLT-1 transporters. When SDR was detected with RH482 dye, which stains both glial and neuronal cells, 1 mM DHK and 1 mM THA were equally effective in suppressing SDR. The SDR was very small in GLT-1 knockout mice but was maintained in gerbil hippocampi in which postsynaptic neurons were absent because of ischemia. Because GLT-1 transporters are exclusively expressed in astrocytes, our results provide direct evidence that astrocytes play the dominant role in sequestering synaptically released glutamate.

Original languageEnglish
Pages (from-to)2580-2588
Number of pages9
JournalJournal of Neuroscience
Volume19
Issue number7
DOIs
Publication statusPublished - 1999 Apr 1

Keywords

  • Astrocytes
  • Brain slice
  • Glutamate transporter
  • Glutamate uptake
  • Hippocampus
  • Voltage-sensitive dye

ASJC Scopus subject areas

  • Neuroscience(all)

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  • Cite this

    Kojima, S., Nakamura, T., Nidaira, T., Nakamura, K., Ooashi, N., Ito, E., Watase, K., Tanaka, K., Wada, K., Kudo, Y., & Miyakawa, H. (1999). Optical detection of synaptically induced glutamate transport in hippocampal slices. Journal of Neuroscience, 19(7), 2580-2588. https://doi.org/10.1523/jneurosci.19-07-02580.1999