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Inhibition of Gap Junction Elevates Glutamate Uptake in Cultured Astrocytes

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Abstract

Glutamate uptake is a main function of astrocytes to keep extracellular glutamate levels low and protect neurons against glutamate-induced excitotoxicity. On the other hand, astrocyte networks formed by gap junctions, which are consisted with connexins and connecting neighboring cells, are reported to play a critical role in maintaining the homeostasis in the brain. In the present study, we examined the effects of gap junction inhibitors on the glutamate uptake activity in cultured rat cortical astrocytes. At first, we confirmed the effects of gap junction inhibitors, 1-octanol and carbenoxolone, on cell–cell communication by the scrape-loading assay using a fluorescent dye Lucifer yellow. Both of 1-octanol and carbenoxolone treatments for 20 min in cultured astrocytes significantly suppressed the cell–cell communication assessed as the distance of dye-spreading. 1-octanol and carbenoxolone increased the glutamate uptake by astrocytes and glutamate aspartate transporter (GLAST) expression on the cell membrane. These results suggest that gap junction inhibitors increase the glutamate uptake activity through the increase of GLAST proteins located on the cell membrane. The regulation of gap junction in astrocytes might protect neurons against glutamate-induced excitotoxicity.

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Abbreviations

CNS:

Central nervous system

Cx:

Connexin

DMEM:

Dulbecco’s modified eagle medium

FBS:

Fetal bovine serum

HHBSS:

HEPES-buffered Hank’s balanced salt solution

HKR:

HEPES-buffered Krebs Ringer solution

HRP:

Horseradish peroxidase

PBS:

Phosphate-buffered saline

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Acknowledgements

This work was supported in part by JSPS KAKENHI Grant Number JP26850209 to K.T., JP26450447 to M.M., JP24621008 to Y.N., and JP15K07768 to Y.N.

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Authors and Affiliations

  1. Laboratory of Integrative Physiology in Veterinary Sciences, Osaka Prefecture University, 1-58, Rinku-Ourai Kita, Izumisano, Osaka, 598-8531, Japan

    Katsura Takano, Masato Ogawa, Kenji Kawabe, Mitsuaki Moriyama & Yoichi Nakamura

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  1. Katsura Takano
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  2. Masato Ogawa
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  3. Kenji Kawabe
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  4. Mitsuaki Moriyama
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  5. Yoichi Nakamura
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Correspondence to Katsura Takano.

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Takano, K., Ogawa, M., Kawabe, K. et al. Inhibition of Gap Junction Elevates Glutamate Uptake in Cultured Astrocytes. Neurochem Res 43, 59–65 (2018). https://doi.org/10.1007/s11064-017-2316-7

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  • DOI: https://doi.org/10.1007/s11064-017-2316-7

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