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To understand the mechanisms of sexual behavior, transcriptomic sequencing technology was used to identify differentially expressed genes (DEGs) in the hypothalamus (HT), pars tuberalis (PT) and pineal gland (PG) in Rasa Aragonesa rams with different sexual behavior. Bioinformatics analysis of the 16,401 identified genes by RNA-Seq revealed 103 and 12 DEGs in the HT and the PG, respectively, at a false discovery rate (FDR) of 5% with an absolute value of expression \u2265 1 (log2FC). However, no DEGs were found in the PT. Functional annotation and pathway enrichment analysis showed that DEGs of HT were enriched mainly in neuroactive ligand-receptor interactions and signaling pathways, including notable candidate genes such as MTNR1A, CHRNA2, FSHB, LHB, GNRHR, AVP, PRL, PDYN, CGA, GABRD, and TSHB, which play a crucial role in sexual behavior. The GnRH and cAMP signaling pathways were also highlighted. In addition, gene set enrichment analysis (GSEA) identified potential pathways, dominated mainly by biological process category, that could be responsible for the differences in sexual behavior observed in rams. The intracellular protein transport and pattern specification process were enriched within the PT and the transcription factor binding and protein ubiquitination pathways for the PG. Thus, these pathways together may play an important role in the regulation of the sexual behavior in Rasa Aragonesa rams through the hypothalamic-pituitary-gonadal axis. The validation of 5 DEGs using reverse transcription quantitative polymerase chain reaction (RT-qPCR) showed expression patterns like the found with RNA-Seq. Overall, these results contribute to understanding the genomic basis of sexual behavior in rams. Our study demonstrates that multiple networks and pathways orchestrate sexual behavior in sheep.<\/jats:p>","DOI":"10.1093\/jas\/skac365","type":"journal-article","created":{"date-parts":[[2022,11,4]],"date-time":"2022-11-04T11:18:26Z","timestamp":1667560706000},"source":"Crossref","is-referenced-by-count":8,"title":["Exploring differentially expressed genes in hypothalamic, <i>pars tuberalis<\/i> and pineal gland transcriptomes in different sexual behavior phenotypes in rams using RNA-Seq"],"prefix":"10.1093","volume":"101","author":[{"given":"Kenza","family":"Lakhssassi","sequence":"first","affiliation":[{"name":"Agrifood Research and Technology Centre of Aragon-IA2 , 50059 Zaragoza , Spain"},{"name":"INRA Instituts , 6356 Rabat , Morocco"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda Pilar","family":"Sarto","sequence":"additional","affiliation":[{"name":"Agrifood Research and Technology Centre of Aragon-IA2 , 50059 Zaragoza , Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bel\u00e9n","family":"Mar\u00edn","sequence":"additional","affiliation":[{"name":"Centre for Encephalopathies and Emerging Transmissible Diseases, Faculty of Veterinary Medicine, University of Zaragoza , 50018 Zaragoza , Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bel\u00e9n","family":"Lahoz","sequence":"additional","affiliation":[{"name":"Agrifood Research and Technology Centre of Aragon-IA2 , 50059 Zaragoza , Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jos\u00e9","family":"Folch","sequence":"additional","affiliation":[{"name":"Agrifood Research and Technology Centre of Aragon-IA2 , 50059 Zaragoza , Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jos\u00e9 Luis","family":"Alabart","sequence":"additional","affiliation":[{"name":"Agrifood Research and Technology Centre of Aragon-IA2 , 50059 Zaragoza , Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Malena","family":"Serrano","sequence":"additional","affiliation":[{"name":"Department of Animal Breeding and Genetics, INIA-CSIC , 28040 Madrid , Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9513-0219","authenticated-orcid":false,"given":"Jorge Hugo","family":"Calvo","sequence":"additional","affiliation":[{"name":"Agrifood Research and Technology Centre of Aragon-IA2 , 50059 Zaragoza , Spain"},{"name":"ARAID , 50018 Zaragoza , Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2022,11,4]]},"reference":[{"key":"2023112010540717600_CIT0001","first-page":"2231","article-title":"Polymorphisms of the melatonin receptor 1A (MTNR1A) gene affect sexual performance of Rasa Aragonesa rams without changing their social dominance","volume":"10","author":"Abecia","year":"2022","journal-title":"J. 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