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Stimulation of norepinephrine transporter function by fasudil, a Rho kinase inhibitor, in cultured bovine adrenal medullary cells

Naunyn-Schmiedeberg's Archives of Pharmacology volume 385, pages 921–931 (2012)Cite this article

Abstract

Norepinephrine transporter (NET) regulates noradrenergic synaptic transmission by controlling extracellular levels of norepinephrine (NE). The small GTPase, RhoA, and its downstream effector Rho kinase (ROCK) are involved in the regulation of actin cytoskeleton and focal adhesion/stress fiber formation, which may play an important role in various functions of the sympathetic nervous system. We report here the effect of fasudil, a ROCK inhibitor, on the functions of NET in cultured bovine adrenal medullary cells as a model of sympathetic neurons. Treatment of bovine adrenal medullary cells with fasudil caused an increase in [3H]NE uptake in time (8–120 h) and concentration (10–100 μM)-dependent manner. Another ROCK inhibitor, Y-27632 (10–100 μM, 1 day), also increased [3H]NE uptake by the cells. Kinetics analysis of the effect of fasudil on NE transport showed a significant increase in the V max of NE transport with little change in K m. When both extracellular and intracellular Ca2+ were removed by the deprivation of extracellular Ca2+ and BAPTA-AM, a cell-permeable Ca2+ chelator, [3H]NE uptake induced by fasudil was completely abolished. Nocodazole, an inhibitor of microtubule polymerization, but not cytochalasin D, an inhibitor of actin polymerization, suppressed the stimulatory effect of fasudil on [3H]NE uptake. The present findings suggest that the ROCK inhibitor fasudil up-regulates NET function in a Ca2+-dependent and/or nocodazole-sensitive pathway in adrenal medullary cells.

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Acknowledgments

This research was supported, in part, by a Grant-in-Aid (23617035, 23590159, and 23617036) for Scientific Research (C) from the Japan Society for the Promotion of Science. We are grateful to Asahi Kasei Corporation (Tokyo, Japan) for providing us with generous supply of fasudil.

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Affiliations

  1. Bio-information Research Center, University of Occupational and Environmental Health, 1-1, Iseigaoka, Yahatanishi-ku, Kitakyushu, 807-8555, Japan

    Noriaki Satoh

  2. Department of Pharmacology, School of Medicine, University of Occupational and Environmental Health, 1-1, Iseigaoka, Yahatanishi-ku, Kitakyushu, 807-8555, Japan

    Yumiko Toyohira & Nobuyuki Yanagihara

  3. Department of Rehabilitation Medicine, School of Medicine, University of Occupational and Environmental Health, 1-1, Iseigaoka, Yahatanishi-ku, Kitakyushu, 807-8555, Japan

    Hideaki Itoh

  4. Institute of Traditional Chinese Medicine Research, Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China

    Han Zhang

  5. Department of Occupational Toxicology, Institute of Industrial Ecological Sciences, University of Occupational and Environmental Health, 1-1, Iseigaoka, Yahatanishi-ku, Kitakyushu, 807-8555, Japan

    Susumu Ueno

  6. Department of Pharmacology, Graduate School of Medicine, University of the Ryukyus, Okinawa, 903-0215, Japan

    Masato Tsutsui

  7. Department of Hospital Pharmacy, University of Occupational and Environmental Health, 1-1, Iseigaoka, Yahatanishi-ku, Kitakyushu, 807-8555, Japan

    Kojiro Takahashi

Authors
  1. Noriaki Satoh
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  2. Yumiko Toyohira
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  3. Hideaki Itoh
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  4. Han Zhang
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  8. Nobuyuki Yanagihara
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Correspondence to Nobuyuki Yanagihara.

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Satoh, N., Toyohira, Y., Itoh, H. et al. Stimulation of norepinephrine transporter function by fasudil, a Rho kinase inhibitor, in cultured bovine adrenal medullary cells. Naunyn-Schmiedeberg's Arch Pharmacol 385, 921–931 (2012). https://doi.org/10.1007/s00210-012-0773-8

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  • DOI: https://doi.org/10.1007/s00210-012-0773-8

Keywords

  • Adrenal medulla
  • Ca2+ dependence
  • Fasudil
  • Norepinephrine transporter
  • Rho kinase inhibitor
  • Up-regulation