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Evolution of the nitric oxide synthase family in vertebrates and novel insights in gill development

Annona, Giovanni and Sato, Iori and Pascual-Anaya, Juan and Braasch, Ingo and Voss, Randal and Stundl, Jan and Soukup, Vladimir and Kuratani, Shigeru and Postlethwait, John H. and D’Aniello, Salvatore (2021) Evolution of the nitric oxide synthase family in vertebrates and novel insights in gill development. . (Unpublished)

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Nitric oxide (NO) is an ancestral key signaling molecule essential for life and has enormous versatility in biological systems, including cardiovascular homeostasis, neurotransmission, and immunity. Although our knowledge of nitric oxide synthases (Nos), the enzymes that synthesize NO in vivo, is substantial, the origin of a large and diversified repertoire of nos gene orthologs in fish with respect to tetrapods remains a puzzle. The recent identification of nos3 in the ray-finned fish spotted gar, which was considered lost in the ray-finned fish lineage, changed this perspective. This prompted us to explore nos gene evolution and expression in depth, surveying vertebrate species representing key evolutionary nodes. This study provides noteworthy findings: first, nos2 experienced several lineage-specific gene duplications and losses. Second, nos3 was found to be lost independently in two different teleost lineages, Elopomorpha and Clupeocephala. Third, the expression of at least one nos paralog in the gills of developing shark, bichir, sturgeon, and gar but not in arctic lamprey, suggest that nos expression in this organ likely arose in the last common ancestor of gnathostomes. These results provide a framework for continuing research on nos genes’ roles, highlighting subfunctionalization and reciprocal loss of function that occurred in different lineages during vertebrate genome duplications.

Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription Paper
Annona, Giovanni0000-0001-7806-6761
Pascual-Anaya, Juan0000-0003-3429-9453
Braasch, Ingo0000-0003-4766-611X
Stundl, Jan0000-0002-3740-3378
Soukup, Vladimir0000-0002-1914-283X
Kuratani, Shigeru0000-0001-9717-7221
Postlethwait, John H.0000-0002-5476-2137
D’Aniello, Salvatore0000-0001-7294-1465
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This version posted June 14, 2021. The authors thank Allyse Ferrara and Quenton Fontenot, Louisiana State University (USA), for their help in the generation of spotted gar embryos. The authors thank Fumiaki Sugahara for his help in the interpretation of results in the arctic lamprey, Anna Pospisilova for technical assistance with bichir and sturgeon in situ hybridizations, and Martin Psenicka, Roman Franek, Michaela Fucikova, Marek Rodina, David Gela, and Martin Kahanec for sterlet sturgeon spawns. A special thanks to Robert Cerny for the establishment of the African bichirs colony at the Charles University in Prague. Giovanni Annona was supported by the Research grant POR Campania FSE 2014/2020 (IT) and by the EMBO Short Term Fellowship (# 6936) to visit the Postlethwait laboratory in Oregon (USA) and for the field trip in Louisiana (USA). Jan Stundl is supported by the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No. 897949. Vladimir Soukup is supported by the Charles University Research Centre program No. 204069 and grant SVV260571/2020. Randal Voss and the Ambystoma Genetic Stock Center are supported by the National Institutes of Health, USA (P40OD019794). John H. Postlethwait is supported by the R01 OD011116 grant from the US National Institutes of Health. Salvatore D’Aniello is supported by the NOEVO grant from the Stazione Zoologica Anton Dohrn Napoli. Data availability: Accession numbers of protein sequences used in the phylogenetic analysis are available in Supplementary Table 1. Primer sequences used for the synthesis of in situ hybridization riboprobes and in quantitative real-time PCR experiments are given in Supplementary Table 2. The authors declare no competing interests.
Funding AgencyGrant Number
Programma Operativo Regionale CampaniaFSE 2014/2020
European Molecular Biology Organization (EMBO)6936
Marie Curie Fellowship897949
Charles University204069
Charles UniversitySVV260571/2020
NIHR01 OD011116
Stazione Zoologica Anton Dohrn NapoliUNSPECIFIED
Record Number:CaltechAUTHORS:20210621-171714685
Persistent URL:
Official Citation:Evolution of the nitric oxide synthase family in vertebrates and novel insights in gill development. Giovanni Annona, Iori Sato, Juan Pascual-Anaya, Ingo Braasch, Randal Voss, Jan Stundl, Vladimir Soukup, Shigeru Kuratani, John H. Postlethwait, Salvatore D’Aniello. bioRxiv 2021.06.14.448362; doi:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:109507
Deposited By: Tony Diaz
Deposited On:21 Jun 2021 17:34
Last Modified:16 Nov 2021 19:36

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