Xi CHEN, Song JIANG, Hong SHEN, Xiancun ZENG, Hanying CHEN

Qira Kara Koyunlarında Östrus Siklusunda LncRNA EkspresyonununBelirlenmesi ve miRNA Analizi İle Kombinasyonu

Bu çalışmada, Qira kara koyunlarının ovaryumlarında lncRNA’ların ekspresyonunun araştırılması için materyal olarak östrus siklusunun farklı aşamalarında olan Qira kara koyunları kullanıldı. Total RNA eldesi amacıyla, östrus senkronizasyonunu takiben siklusun dört farklı aşamasından ovaryum dokuları toplandı. Farklı aşamalardaki örnekler, önce RNA-seq teknolojisi kullanılarak genom çapında analize tabi tutuldu, lncRNA’ların hedef genleri birlikte ifade edilme yöntemleri kullanılarak tahmin edildi ve takiben hedef genlerin analizi GO ve KEGG ile gerçekleştirildi. Qira kara koyunlarında östrus siklusu ile ilgili genler, transkripsiyonel farklılıklarının araştırılması amacıyla seçildi. Son olarak, lncRNA hizalamasının Qira kara koyunlarında üreme üzerine etkisini daha fazla analiz etmek için lncRNA-miRNA ve mRNA-lncRNA-miRNA etkileşim ağları kuruldu. Sonuçlar, çeşitli ekspresyon bölgelerinde, 14 lncRNA’nın diöstrusa göre östrusta farklı şekillerde eksprese edildiğini gösterdi ve metöstrusa karşı östrus ve proöstrusa karşı östrus gruplarında lncRNA’ların diferansiyel ekspresyon seviyeleri sırasıyla 18 ve 24 saptandı. GO ve KEGG fonksiyonel zenginleştirme analizleri, farklı şekilde eksprese edilen lncRNA’ların ve bunların hedef genlerinin, esas olarak retinol metabolizması, yumurtalık sterol üretimi ve endosterol biyosentezi gibi üreme ile ilgili fonksiyonlarda yer aldığını gösterdi. lncRNA-miRNA ve mRNA-lncRNA-miRNA’nın kombine analizi sonucu, LNC011583, LNC003443 ve bta-miR-202 gibi üreme ile ilgili genler saptandı, dolayısıyla lncRNA’ların Qira kara koyunlarının üremesinde bir miktar etkisinin olduğu görülebilir.

Identification of LncRNA Expression in the Estrous Cycle of Qira Black Sheep and Its Combination with miRNA Analysis

In order to investigate the expression of lncRNAs in the ovaries of Qira black sheep at different stages of the estrous cycle, Qira black sheep were used as experimental materials in this experiment, and after estrus synchronization, ovarian tissues at four different stages of the estrous cycle were collected to extract total RNA.The samples at different stages were first subjected to genome-wide analysis using RNA-seq technology; target genes of lncRNAs were predicted using co-expression methods; and then GO and KEGG analysis of target genes was performed. Genes related to the estrous cycle of Qira black sheep were also selected to study their transcriptional differences. Finally, lncRNA-miRNA and mRNAlncRNA-miRNA interaction networks were established to further analyze the effect of lncRNA alignment on Qira black sheep reproduction. The results showed that in the differentially expressed part, 14 lncRNAs were differentially expressed in Estrous VS Diestrus; the differential expression levels of lncRNAs in the two comparison groups of Estrous VS Metestrus and Estrus VS Proestrus were 18 and 24, respectively. The results of GO and KEGG functional enrichment analysis showed that differentially expressed lncRNAs and their target genes were mainly involved in reproductionrelated pathways such as retinol metabolism, ovarian sterol production and endosterol biosynthesis. In the combined analysis of lncRNA-miRNA and mRNA-lncRNA-miRNA, genes related to reproduction, such as LNC011583, LNC003443 and bta-miR-202, were found, thus it can be seen that lncRNAs have some effect in the reproduction of Qira black sheep.

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