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Adad1 mRNA is expressed in both sexes and is germline specific. A-D: RT-PCR assaying adad1 expression in wild-type tissues. RT-PCR on adult tissues demonstrates that adad1 is expressed only in ovary and testis (A), whereas the control gene rpl13a is expressed in all organs tested (B). During embryonic and larval development adad1 is expressed in all stages tested (C), similar to the germ cell marker piwil1 (D). E: RT-PCR on testes lacking germ cells revealed that adad1 is not expressed in somatic cells of the testis. Piwil1 expression was not detected confirming that germ cells are not present while the Leydig cell marker gene cyp11c1 confirmed the presence of testes tissue. F-K: In situ hybridization (ISH) and single cell RNA sequencing data showing adad1 is expressed in germ cells of adult ovaries and testes. (F-H) In testes adad1 expression was highest in spermatogonia, spermatocytes, and less expressed in early spermatids. Testes were counterstained with periodic acid-Schiff. (I-K) In ovaries adad1 expression was found in oogonia through early follicle stages. Scale bars: 20 μm for testes and 50 μm for ovaries. Abbreviations: SPG1 = spermatogonia cluster 1; SPG2 = spermatogonia cluster 2; SPC = spermatocytes; E_Round = early round spermatids; M_Round = middle round spermatids; L_Round = late round spermatids; Elongated = elongated spermatids; GSC_Pro = germline stem cell and progenitor cells; E_Meio = early meiosis, Meio = Meiosis, L_Meiosis = Late meiosis; E_Oo1 = early follicle stage oocyte cluster 1; E_Oo2 = early follicle stage oocyte cluster 2; E_Oo3 = early follicle stage oocyte cluster 3; Follicle_Lhx9 = Lx9-expressing follicle cells; Follicle1 = follicle cells cluster 1; Follicle2 = Follicle cells cluster 2; Theca = Theca cells.
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