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Ant queens and workers vary in fecundity and lifespan, which are enhanced and extended in queens. Yet, the regulatory mechanisms underlying this variation are not well understood. Ant queens live and reproduce for years, so that they need to protect their germline from transposable element (TE) activity, which may be redundant in short\u2010lived, often sterile workers. We analysed the expression of two protective classes of small RNAs, microRNAs (miRNAs) and Piwi\u2010interacting RNAs (piRNAs), in various tissues, castes and age classes of the ant <jats:italic>Temnothorax rugatulus<\/jats:italic>. In queens, piRNAs were highly abundant in ovaries with TEs being their clear targets, with reduced but still detectable piRNA\u2010specific ping\u2010pong signatures in thorax and brains. piRNA pathway activity varied little with age in queens. Moreover, the reduced ovaries of workers also exhibited similar piRNA activity and this not only in young, fertile workers, but also in older foragers with regressed ovaries. Therefore, these ants protect their germline through piRNA activity, regardless of ovarian development, age or caste, even in sterile workers often considered the soma of the <jats:italic>superorganism<\/jats:italic>. Our tissue\u2010specific miRNA analysis detected the expression of 304 miRNAs, of which 105 were expressed in all tissues, 10 enriched in the brain, three in the thorax, whereas 83 were ovarian\u2010specific. We identified ovarian miRNAs whose expression was related to caste, fecundity and age, and which likely regulate group\u2010specific gene expression. sRNA shifts in young\u2010 to middle\u2010aged queens were minor, suggesting delayed senescence in this reproductive caste.<\/jats:p>","DOI":"10.1111\/mec.17162","type":"journal-article","created":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T11:26:11Z","timestamp":1697196371000},"page":"6027-6043","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Age\u2010 and caste\u2010independent <scp>piRNAs<\/scp> in the germline and <scp>miRNA<\/scp> profiles linked to caste and fecundity in the ant <i>Temnothorax rugatulus<\/i>"],"prefix":"10.1111","volume":"32","author":[{"given":"Ann\u2010Sophie","family":"Seistrup","sequence":"first","affiliation":[{"name":"Institute of Molecular Biology  Mainz Germany"},{"name":"International PhD Programme on Gene Regulation, Epigenetics &amp; Genome Stability  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Choppin","sequence":"additional","affiliation":[{"name":"Institute of Organismic and Molecular Evolution Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shamitha","family":"Govind","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology  Mainz Germany"},{"name":"International PhD Programme on Gene Regulation, Epigenetics &amp; Genome Stability  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0413-7245","authenticated-orcid":false,"given":"Barbara","family":"Feldmeyer","sequence":"additional","affiliation":[{"name":"Senckenberg Biodiversity and Climate Research Centre (SBiK\u2010F), Molecular Ecology  Frankfurt Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marion","family":"Kever","sequence":"additional","affiliation":[{"name":"Institute of Organismic and Molecular Evolution Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emil","family":"Karaulanov","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alice","family":"S\u00e9guret","sequence":"additional","affiliation":[{"name":"School of Biological Sciences University of Bristol  Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sivarajan","family":"Karunanithi","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel V.","family":"Almeida","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology  Mainz Germany"},{"name":"Department of Biochemistry University of Cambridge  Cambridge UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ren\u00e9 F.","family":"Ketting","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology  Mainz Germany"},{"name":"Institute of Developmental Biology and Neurobiology Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8161-6306","authenticated-orcid":false,"given":"Susanne","family":"Foitzik","sequence":"additional","affiliation":[{"name":"Institute of Organismic and Molecular Evolution Johannes Gutenberg University Mainz  Mainz Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,10,13]]},"reference":[{"key":"e_1_2_9_2_1","unstructured":"Andrews S.(2010).FASTQC. 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