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Epigenetic Features Induced by Ischemia-Hypoxia in Cultured Rat Astrocytes

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Abstract

Epigenetic mechanisms are involved in the maintenance of long-term hypoxia-adapted cellular functions. Astrocytes as the dividing neuroglia cell in the nervous tissue can be activated under hypoxia condition. In the present study, the epigenetic characteristics such as DNA methylation and histone acetylation, as well as S-adenosylmethionine (SAM) role were explored in cultured astrocytes under ischemia-hypoxia (IH) condition. IH could induce the hypermethylation of global DNA and hypoacetylation of histone H3/H4 in an enzyme-dependent manner. c-Jun amino terminal kinase (JNK) signal pathway was involved in this epigenetic change induced by IH. SAM revealed a reverse effect on protein expression without the involvement of methyl donor. On the other hand, neither change of global DNA nor specific gene methylation level was observed in non-dividing neurons under IH condition. In conclusion, a genome-wide adjustment of DNA methylation and histone acetylation under IH conditions should be involved in epigenetic programming of astrocytes. Understanding how ischemia-hypoxia affects global and gene-specific epigenetic programming will provide important insights into the mechanisms of hypoxia-induced cellular changes.

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Acknowledgments

This work was supported by National Natural Science Foundation (No. 81070926 and No. 81271310).

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The authors declare no conflict of interests.

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Authors and Affiliations

  1. Department of Pathology, Key Laboratory for Neurodegenerative Disease of Education Ministry, Capital Medical University, Beijing, 100069, China

    Qinglin Yang, Xiangmei Wu, Jing Sun, Jing Cui & Liang Li

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Correspondence to Liang Li.

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Supplementary Fig. 1

Schematic diagram of APP and BACE1 genes. The transcripts were composed of a 5’-untranslated region (UTR) with internal promoter activity, open reading frame (ORF), and a 3’-UTR. (A) The sequence represents a 240-bp fragment (between 26 and 266) in APP gene. (B) The sequence represents a 250-bp fragment (between -318 and -58) in BACE1 gene. The number refers to the location of cytosine-guanine (CpG) sites and underlying highlights of CpG sites include more than 1 CpG site tested at the same time. (GIF 136 kb)

High Resolution Image (TIFF 2716 kb)

Supplementary Fig. 2

Micrographs of cultured astrocytes and neurons. (A) Representative cultured astrocytes were immunocytochemically stained against GFAP (green). (B) Representative cultured neurons were immunocytochemically stained against Tuj1 (green). Scale bar = 50 μm. (GIF 71 kb)

High Resolution Image (TIFF 4917 kb)

Supplementary Fig. 3

Evaluation of cell viability for cultured cells under IH treatment. The cultured cells subjected to IH treatment were double stained by Hoechst 33258 (blue) and PI (red). (A) Control, (B) IH treatment for 24 h, (C) IH treatment for 48 h, (D) IH treatment for 72 h. Scale bar = 100 μm. (E) The number of dead cells after IH treatment. The data were expressed as mean ± SD. *p = < 0.05; **p < 0.01 vs. control group (n = 3). (GIF 147 kb)

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Yang, Q., Wu, X., Sun, J. et al. Epigenetic Features Induced by Ischemia-Hypoxia in Cultured Rat Astrocytes. Mol Neurobiol 53, 436–445 (2016). https://doi.org/10.1007/s12035-014-9027-8

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