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Gaining a deeper understanding of its reproductive biology offers valuable insights into its evolutionary success and may inform the development of sustainable strategies to control its spread. This study presents a comprehensive structural and functional characterization of the fruitless gene in Ae. albopictus (Aalfru), a pivotal regulator of sexual behavior in insects. Through in silico analysis combined with molecular and functional genetics approaches, we identified a high degree of conservation in the fru gene structure and its regulation via sex-specific alternative splicing. Differently from Drosophila, Aedes aegypti, and other dipteran fruitless orthologs, Aalfru sex-specific regulation starts in 1-day-old embryos, rather than the late larval stage. Functional analysis using embryonic RNA interference (RNAi) demonstrated that, Ae. albopictus males with transiently disrupted fru expression at the embryonic stage showed significant deficits in adult mating behavior and failed to produce viable progeny. Our findings elucidate the Aalfru gene\u2019s molecular organization, developmental regulation, and critical role in courtship behavior, highlighting its importance in male sexual behavior and reproductive success in Ae. albopictus.<\/jats:p>","DOI":"10.3390\/insects16030280","type":"journal-article","created":{"date-parts":[[2025,3,7]],"date-time":"2025-03-07T12:22:52Z","timestamp":1741350172000},"page":"280","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The Early Sex-Specific Expression of the Fruitless Gene in the Asian Tiger Mosquito Aedes albopictus (Skuse) and Its Functional Conservation in Male Courtship"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-3831-7074","authenticated-orcid":false,"given":"Marianna","family":"Varone","sequence":"first","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3181-3960","authenticated-orcid":false,"given":"Paola","family":"Di Lillo","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4194-0368","authenticated-orcid":false,"given":"Katerina","family":"Nikolouli","sequence":"additional","affiliation":[{"name":"Insect Pest Control Laboratory, Joint FAO\/IAEA Centre of Nuclear Techniques in Food and Agriculture, Department of Nuclear Sciences and Applications, IAEA Laboratories, 2444 Seibersdorf, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayca Eda","family":"\u00d6zel","sequence":"additional","affiliation":[{"name":"Insect Pest Control Laboratory, Joint FAO\/IAEA Centre of Nuclear Techniques in Food and Agriculture, Department of Nuclear Sciences and Applications, IAEA Laboratories, 2444 Seibersdorf, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7597-0288","authenticated-orcid":false,"given":"Francesca","family":"Lucibelli","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5274-5854","authenticated-orcid":false,"given":"Gennaro","family":"Volpe","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3355-4827","authenticated-orcid":false,"given":"Sarah Maria","family":"Mazzucchiello","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-9715-7786","authenticated-orcid":false,"given":"Angela","family":"Carfora","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3906-4872","authenticated-orcid":false,"given":"Serena","family":"Aceto","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9835-3693","authenticated-orcid":false,"given":"Giuseppe","family":"Saccone","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kostas","family":"Bourtzis","sequence":"additional","affiliation":[{"name":"Insect Pest Control Laboratory, Joint FAO\/IAEA Centre of Nuclear Techniques in Food and Agriculture, Department of Nuclear Sciences and Applications, IAEA Laboratories, 2444 Seibersdorf, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Salvemini","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Study of Naples Federico II, 80100 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.anbehav.2021.03.017","article-title":"Physiological innovation and the evolutionary elaboration of courtship behaviour","volume":"184","author":"Fuxjager","year":"2022","journal-title":"Anim. 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