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Ammonia and Manganese Increase Arginine Uptake in Cultured Astrocytes

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Abstract

Recent work has suggested a possible role for nitric oxide (NO) in the development of hepatic encephalopathy (HE). In this study, we examined the effect of ammonia and manganese, factors implicated in the pathogenesis of HE, on the transport of arginine (a precursor of NO) into primary cultures of astrocytes. Treatment with 5 mM ammonia for 1–4 days produced a maximal (53%) increase in L-arginine uptake at 3 days when compared to untreated cells. Kinetic analysis following 4-day treatment with 5 mM ammonia revealed an 82% increase in the Vmax and a 61% increase in the Km, value. Similar analysis with 100 μM manganese showed a 101% increase in Vmax and a 131% increase in the Km value. These results suggest that both manganese and ammonia alter L-arginine uptake by modifying the transporter for arginine. A decrease of 32% in the non-saturable component of L-arginine transport was also observed following treatment with ammonia. When cultures were treated separately with 5 mM ammonia and 100 μM manganese for 2 days, the uptake of L-arginine increased by 41% and 57%, respectively. Combined exposure led to no further increase in uptake. Our results suggest that ammonia and manganese may contribute to the pathogenesis of HE by influencing arginine transport and thus possibly NO synthesis in astrocytes.

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Authors and Affiliations

  1. Laboratory of Neuropathology, Veterans Administration Medical Center, USA

    Alan S. Hazell & Michael D. Norenberg

  2. University of Miami School of Medicine, Miami, FL, 33101

    Alan S. Hazell

  3. Department of Biochemistry & Molecular Biology, University of Miami School of Medicine, Miami, FL, 33101

    Michael D. Norenberg

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Hazell, A.S., Norenberg, M.D. Ammonia and Manganese Increase Arginine Uptake in Cultured Astrocytes. Neurochem Res 23, 869–873 (1998). https://doi.org/10.1023/A:1022411012512

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