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The effects of the neurosteroids: pregnenolone, progesterone and dehydroepiandrosterone on muscarinic receptor-induced responses in Xenopus oocytes expressing M1 and M3 receptors

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Abstract

The neurosteroids pregnenolone, progesterone, and dehydroepiandrosterone (DHEA) occur naturally in the nervous system. They act on neural tissues, participate in neuronal signaling, and are reported to alter neuronal excitability via nongenomic mechanisms. Muscarinic receptors have important roles in neuronal functions in the brain and autonomic nervous system. In this study, we investigated the effects of pregnenolone, progesterone, and DHEA on M1 and M3 muscarinic receptors using the Xenopus oocyte expression system. Pregnenolone and progesterone inhibited the acetylcholine (ACh)-mediated responses of M1 and M3 receptors expressed in Xenopus oocytes, whereas DHEA did not. The half-maximal inhibitory concentrations (IC50) for pregnenolone inhibition of M1 receptor- and M3 receptor-mediated currents were 11.4 and 6.0 μM respectively; the IC50 values for progesterone inhibition of M1 receptor- and M3 receptor-mediated currents were 2.5 and 3.0 μM respectively. The selective protein kinase C (PKC) inhibitor GF109203X had little effect on the pregnenolone or progesterone inhibition of the ACh-induced currents in Xenopus oocytes expressing M1 or M3 receptors. The inhibitory effects of pregnenolone and progesterone were overcome at higher concentrations of ACh. Pregnenolone and progesterone inhibited the [3H]quinuclidinyl benzilate (QNB) binding to M1 and M3 receptor expressed in Xenopus oocytes, and Scatchard plot analysis of [3H]QNB binding revealed that pregnenolone and progesterone altered the Kd value and the Bmax, indicating noncompetitive inhibition. In conclusion, pregnenolone and progesterone inhibited M1 and M3 receptor functions noncompetitively by the mechanism independent of PKC and by interfering with ACh binding to the receptors.

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Authors and Affiliations

  1. Department of Anesthesiology, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishiku, Kitakyushu, 807-8555, Japan

    Takafumi Horishita, Kouichiro Minami, Munehiro Shiraishi, Junichi Ogata, Takashi Okamoto, Tadanori Terada & Takeyoshi Sata

  2. Department of Pharmacology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan

    Yasuhito Uezono

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  1. Takafumi Horishita
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  2. Kouichiro Minami
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  3. Yasuhito Uezono
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  4. Munehiro Shiraishi
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  5. Junichi Ogata
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  6. Takashi Okamoto
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  7. Tadanori Terada
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  8. Takeyoshi Sata
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Correspondence to Kouichiro Minami.

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Horishita, T., Minami, K., Uezono, Y. et al. The effects of the neurosteroids: pregnenolone, progesterone and dehydroepiandrosterone on muscarinic receptor-induced responses in Xenopus oocytes expressing M1 and M3 receptors. Naunyn-Schmiedeberg's Arch Pharmacol 371, 221–228 (2005). https://doi.org/10.1007/s00210-005-1022-1

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