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Ammonium Increases Ca2+ Signalling and Up-Regulates Expression of TRPC1 Gene in Astrocytes in Primary Cultures and in the In Vivo Brain

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Abstract

Rapid rise in ammonium concentration in the brain is the major pathogenic factor in hepatic encephalopathy that is manifested by state of confusion, forgetfulness and irritability, psychotic symptoms, delusions, lethargy, somnolence and, in the terminal stages, coma. Primary cultures of mouse astrocytes were used to investigate effects of chronic treatment (3 days) with ammonium chloride (ammonium) at 3 mM, this being a relevant concentration for hepatic encephalopathy condition, on metabotropic receptor agonist-induced increases in free cytosolic Ca2+ concentration [(Ca2+)i], measured with fura-2 based microfluorimetry and on store-operated Ca2+ entry (SOCE) activated following treatment with the SERCA inhibitor thapsigargin. The agonists used were the β-adrenergic agonist isoproterenol, the α2-adrenergic agonist dexmedetomidine, the InsP3 receptor (InsP3R) agonist adenophostin A and ryanodine receptor agonist 4-Chloro-m-cresol (4-CMC). Agonist-induced [Ca2+]i responses were significantly increased in astrocytes chronically exposed to ammonium. Similarly, the SOCE, meditated by the transient receptor potential channel 1 (TRPC1), was significantly augmented. The ammonium-induced increase in SOCE was a result of an up-regulation of mRNA and protein expression of TRPC1 in astrocytes. Increase in TRPC1 expression and in SOCE were both prevented by ouabain antagonist canrenone. Similar up-regulation of TRPC1 gene expression was found in the brain of adult mice subjected to intraperitoneal injection of urease for 3 days. In transgenic mice tagged with an astrocyte-specific or a neurone-specific markers and treated with intraperitoneal injections of urease for 3 days, the fluorescence-activated cell sorting of neurones and astrocytes demonstrated that TRPC1 mRNA expression was up-regulated in astrocytes, but not in neurones.

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Acknowledgments

This study was supported by Grants No. 31171036 to L.P. and No. 31300883 to T.D. from the National Natural Science Foundation of China.

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Authors and Affiliations

  1. Laboratory of Metabolic Brain Diseases, Institute of Metabolic Disease Research and Drug Development, China Medical University, No. 92 Beier Road, Heping District, Shenyang, People’s Republic of China

    Chunguang Liang, Ting Du, Jing Zhou & Liang Peng

  2. Faculty of Life Science, The University of Manchester, Manchester, UK

    Alexei Verkhratsky

  3. Achucarro Center for Neuroscience, IKERBASQUE, Basque Foundation for Science, 48011, Bilbao, Spain

    Alexei Verkhratsky

  4. University of Nizhniy Novgorod, Nizhniy Novgorod, 603022, Russia

    Alexei Verkhratsky

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  1. Chunguang Liang
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  2. Ting Du
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  4. Alexei Verkhratsky
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  5. Liang Peng
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Correspondence to Liang Peng.

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Chunguang Liang and Ting Du contributed equally to this article.

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Liang, C., Du, T., Zhou, J. et al. Ammonium Increases Ca2+ Signalling and Up-Regulates Expression of TRPC1 Gene in Astrocytes in Primary Cultures and in the In Vivo Brain. Neurochem Res 39, 2127–2135 (2014). https://doi.org/10.1007/s11064-014-1406-z

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  • DOI: https://doi.org/10.1007/s11064-014-1406-z

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