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Inhibition of Reactive Astrocytes with Fluorocitrate Ameliorates Learning and Memory Impairment Through Upregulating CRTC1 and Synaptophysin in Ischemic Stroke Rats

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Abstract

Ischemic stroke often causes motor and cognitive deficits. Deregulated glia gap junction communication, which is reflected by increased protein levels of glial fibrillary acidic protein (GFAP) and connexin 43 (Cx43), has been observed in ischemic hippocampus and has been associated with cognitive impairment in animal stroke models. Here, we tested the hypothesis that reactive astrocytes-mediated loss of synaptophysin (SYP) and CREB-regulated transcription coactivator 1 (CRTC1) contribute to dysfunction in glia gap junction communication and memory impairment after ischemic stroke. Male Sprague–Dawley rats were subjected to a 90-min middle cerebral artery occlusion (MCAO) with 7-day reperfusion. Fluorocitrate (1 nmol), the reversible inhibitor of the astrocytic tricarboxylic acid cycle, was injected into the right lateral ventricle of MCAO rats once every 2 days starting immediately before reperfusion. The Morris water maze was used to assess memory in conjunction with western blotting and immunostaining to detect protein expression and distribution in the hippocampus. Our results showed that ischemic stroke caused significant memory impairment accompanied by increased protein levels of GFAP and Cx43 in hippocampal tissue. In addition, the levels of several key memory-related important proteins including SYP, CRTC1, myelin basic protein and high-mobility group-box-1 were significantly reduced in the hippocampal tissue. Of note, inhibition of reactive astrocytes with fluorocitrate was shown to significantly reverse the above noted changes induced by ischemic stroke. Taken together, our findings demonstrate that inhibiting reactive astrocytes with fluorocitrate immediately before reperfusion may protect against ischemic stroke-induced memory impairment through the upregulation of CRTC1 and SYP.

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Abbreviations

CRTC1:

CREB-regulated transcription coactivator 1

Cx43:

Connexin 43

FC:

Fluorocitrate

GFAP:

Glial fibrillary acidic protein

GJC:

Glia gap junction communication

HMGB1:

High-mobility group-box-1

MBP:

Myelin basic protein

MCAO:

Middle cerebral artery occlusion

SYP:

Synaptophysin

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Funding

This work was supported, by National Natural Science Foundation of China (81870973, 81671145, 81701316, 81873747), by Jiangsu Provincial College of Natural Science research project (17KJB180012), by Suzhou Science and Technology for People’s Livelihood (SYS2018025), by Natural Science Foundation of Guangdong Province (2016A030313027), by grants from Shenzhen Science & Technology Commission grants (JCYJ20170306093243010 and JCYJ20170413165705083).

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  1. Xinyu Zhang, Xianzhi Shen, Jiali Dong and Wen-Cao Liu have contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology and Suzhou Clinical Research Center of Neurological Disease, Jiangsu Key Laboratory of Neuropsychiatric Diseases, The Second Affiliated Hospital of Soochow University, Institute of Neuroscience, Soochow University, Suzhou, China

    Xinyu Zhang, Xianzhi Shen, Jiali Dong, Min Song, Yanyun Sun, Hui Shu, Chun-Feng Liu & Xinchun Jin

  2. Department of Emergency, Shanxi Provincial People’s Hospital, Taiyuan, China

    Wen-Cao Liu

  3. School of Chemistry and Biosciences, University of Bradford, Bradford, BD7 1DP, UK

    Clare-Louise Towse

  4. The Central Laboratory, Shenzhen Second People’s Hospital, Shenzhen University 1st Affiliated Hospital, Shenzhen University School of Medicine, Shenzhen, 518035, China

    Wenlan Liu

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  1. Xinyu Zhang
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Contributions

This work was performed and accomplished by all authors. XZ, XS, JD, WCL, MS, YS, and HS contributed to the execution of the entire research project and the statistical analyses. XZ, XS, JD, WCL, CLT, WL, CFL, and XJ wrote the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Wenlan Liu, Chun-Feng Liu or Xinchun Jin.

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Zhang, X., Shen, X., Dong, J. et al. Inhibition of Reactive Astrocytes with Fluorocitrate Ameliorates Learning and Memory Impairment Through Upregulating CRTC1 and Synaptophysin in Ischemic Stroke Rats. Cell Mol Neurobiol 39, 1151–1163 (2019). https://doi.org/10.1007/s10571-019-00709-0

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