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Acetylcholine inhibits nitric oxide (NO) synthesis in the gastropod nervous system

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Abstract

Acetylcholine (ACh) is one of the main signals regulating nitric oxide synthase (NOS) expression and nitric oxide (NO) biosynthesis in mammals. However, few comparative studies have been performed on the role of ACh on NOS activity in non-mammalian animals. We have therefore studied the cholinergic control of NOS in the snail Helix pomatia and compared the effects of ACh on NO synthesis in the enteric nervous system of the snail and rat. Analyses by the NADPH-diaphorase reaction, immunocytochemistry, purification with ion-exchange chromatography, Western-blot, and quantitative polymerase chain reaction have revealed the expression of neuronal NOS in the rat intestine and of a 60-kDa subunit of NOS in the enteric nerve plexus of H. pomatia. In H. pomatia, quantification of the NO-derived nitrite ions has established that NO formation is confined to the NOS-containing midintestine. Nitrite production can be elevated by L-arginine but inhibited by Nω-nitro-L-arginine. In rats, ACh moderately elevates nitrite production, whereas ACh, the nicotinic receptor agonists (nicotine, acetyl thiocholine iodide, metacholine) and the cholinesterase inhibitor eserine reduce enteric nitrite formation in snails. The nicotinic receptor antagonist tubocurarine also provokes nitrite liberation, whereas the muscarinic receptor agonists or antagonists have no significant effect in snails. In the presence of EDTA or tetrodotoxin, ACh fails to inhibit nitrite production. In pharmacological studies, we have found that ACh contracts the midintestinal muscles and, in snails, simultaneously reduces the antagonistic muscle relaxant effect of L-arginine. Our experiments provide the first evidence for an inhibitory regulation of neuronal NO synthesis by ACh in an invertebrate species.

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Acknowledgements

The kind help of Dr. Attila Szanto with the RT-QPCR analyses and the technical assistance of Ms. Krisztina Dobák and Ms. Ildikó Juhász is gratefully acknowledged.

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  1. Tamás Rőszer

    Present address: Research Group of Apoptosis & Genomics, Hungarian Academy of Sciences, 4028, Debrecen Pf. 6., Hungary

Authors and Affiliations

  1. Department of Animal Anatomy and Physiology, Faculty of Science, Debrecen University, Debrecen, Hungary

    Tamás Rőszer & Gáspár Bánfalvi

  2. Department of Pediatrics, Medical and Health Science Center, Debrecen University, Debrecen, Hungary

    Tamás Józsa

  3. Department of Pharmacology and Pharmacotherapy, Medical and Health Science Center, Debrecen University, Debrecen, Hungary

    A. József Szentmiklósi

  4. Department of Microbial Biotechnology and Cell Biology, Debrecen University, Debrecen, Hungary

    Tamás Rőszer & Gáspár Bánfalvi

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  1. Tamás Rőszer
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Correspondence to Tamás Rőszer.

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This article is dedicated to Dr. Gábor Hollósi on the 50th anniversary of his graduation and being a teacher at the University of Debrecen.

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Rőszer, T., Józsa, T., Szentmiklósi, A.J. et al. Acetylcholine inhibits nitric oxide (NO) synthesis in the gastropod nervous system. Cell Tissue Res 336, 325–335 (2009). https://doi.org/10.1007/s00441-009-0764-3

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