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Many seasonal challenges require extensive physiological modification to survive. In winter, to survive extreme cold and limited resources, insects commonly enter diapause, which is an endogenously derived dormant state associated with minimized cellular processes and low energetic expenditure. Due to the high degree of complexity involved in diapause, substantial cellular regulation is required, of which our understanding primarily derives from the transcriptome via messenger RNA expression dynamics. Here we aim to advance our understanding of diapause by investigating microRNA (miRNA) expression in diapausing and direct developing pupae of the butterfly <jats:italic>Pieris napi.<\/jats:italic> We identified coordinated patterns of miRNA expression throughout diapause in both head and abdomen tissues of pupae, and via miRNA target identification, found several expression patterns to be enriched for relevant diapause\u2010related physiological processes. We also identified two candidate miRNAs, miR\u201014\u20105p and miR\u20102a\u20103p, that are likely involved in diapause progression through their activity in the ecdysone pathway, a critical regulator of diapause termination. miR\u201014\u20105p targets <jats:italic>phantom<\/jats:italic>, a gene in the ecdysone synthesis pathway, and is upregulated early in diapause. miR\u20102a\u20103p has been found to be expressed in response to ecdysone, and is upregulated during diapause termination. Together, the expression patterns of these two miRNAs match our current understanding of the timing of hormonal regulation of diapause in <jats:italic>P.\u2009napi<\/jats:italic> and provide interesting candidates to further explore the mechanistic role of microRNAs in diapause regulation.<\/jats:p>","DOI":"10.1111\/mec.17348","type":"journal-article","created":{"date-parts":[[2024,4,10]],"date-time":"2024-04-10T07:19:05Z","timestamp":1712733545000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["A time course analysis through diapause reveals dynamic temporal patterns of <scp>microRNAs<\/scp> associated with endocrine regulation in the butterfly <i>Pieris napi<\/i>"],"prefix":"10.1111","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2785-5108","authenticated-orcid":false,"given":"Kevin T.","family":"Roberts","sequence":"first","affiliation":[{"name":"Department of Zoology Stockholm University  Stockholm Sweden"},{"name":"Zoological Institute and Museum, University of Greifswald  Greifswald Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel A.","family":"Steward","sequence":"additional","affiliation":[{"name":"Department of Zoology Stockholm University  Stockholm Sweden"},{"name":"Department of Biology Lund University  Lund Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip","family":"S\u00fcess","sequence":"additional","affiliation":[{"name":"Department of Zoology Stockholm University  Stockholm Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8344-6830","authenticated-orcid":false,"given":"Philipp","family":"Lehmann","sequence":"additional","affiliation":[{"name":"Department of Zoology Stockholm University  Stockholm Sweden"},{"name":"Zoological Institute and Museum, University of Greifswald  Greifswald Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1863-2340","authenticated-orcid":false,"given":"Christopher W.","family":"Wheat","sequence":"additional","affiliation":[{"name":"Department of Zoology Stockholm University  Stockholm Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,4,10]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-018-1504-3"},{"key":"e_1_2_10_3_1","unstructured":"Alexa A. &Rahnenfuhrer J.(2020).TOPGO: Enrichment analysis for gene ontology 2.40. 0. 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