lnc94638 is a testis‐specific long non‐coding RNA involved in spermatozoa formation in Zeugodacus cucurbitae (Coquillett)

Abstract

Long non‐coding RNAs (lncRNAs) generally display tissue‐specific distributions, and testis‐specific lncRNAs form the highest proportion of lncRNAs in many species. Here, we presented a detailed analysis of testis‐specific lncRNAs in the melon fly, Zeugodacus cucurbitae, a highly destructive insect pest of cucurbitaceous and other related crops. Most testis‐specific lncRNAs were found to be long intergenic non‐coding RNAs (lincRNA). The size distribution of these lncRNAs ranged between 600 and 1000 nucleotides. Testis‐specific lncRNAs that harboured one isoform number and two exons were the most abundant. Compared to other male tissues, the testis had more highly expressed lncRNAs. The quantitative real‐time polymerase chain reaction results of 10 randomly selected testis‐specific lncRNAs showed expression patterns consistent with RNA‐seq data. Further analysis of the most highly expressed testis‐specific lncRNA, lnc94638, was undertaken. Fluorescent in situ hybridization assays localized lnc94638 to the apical region of the testis that contains mature spermatozoa. RNA interference‐mediated knockdown of lnc94638 expression reduced spermatozoa numbers and impaired the fertility of Z. cucurbitae male. This study provides a catalogue of testis‐specific lncRNAs, shows that the testis‐specific lnc94638 is involved in spermatogenesis and has the potential to be used for treating male sterility.

Comparative multi‐tissue analyses identify testis‐specific serine/threonine protein kinase (TSSK) genes involved in male fertility in the melon fly Zeugodacus cucurbitae

“…FISH analyses further confirmed that the ZcTSSK1 and ZcTSSK3 signal was largely confined to the transformation region where spermatids finalize their differentiation into tailed spermatozoa. The testis‐specific gene lnc94638 is similarly expressed primarily within this transformation region, and knocking down lnc94638 has been shown to impact male melon fly fertility 44 . As such, both the localization and expression patterns of ZcTSSK1 and ZcTSSK3 strongly suggest that they may function similarly as important mediators of spermatogenesis in Z. cucurbitae .…”

“…The testisspecific gene lnc94638 is similarly expressed primarily within this transformation region, and knocking down lnc94638 has been shown to impact male melon fly fertility. 44 As such, both the localization and expression patterns of ZcTSSK1 and ZcTSSK3 strongly suggest that they may function similarly as important mediators of spermatogenesis in Z. cucurbitae. Consistent findings have also been reported in mammals.…”

lncRNA 1700101O22Rik and NONMMUG030480.1 Are Not Essential for Spermatogenesis in Mice

“…Although lncRNAs have been considered as a “redundant factor” in the individual regulatory system in previous studies, more evidence has shown that lncRNAs can direct or indirectly participate in cell function regulation and the occurrence of various human diseases [ 29 ]. Accumulating studies have characterized many testis-related lncRNAs [ 30 , 31 , 32 ], but only some pieces of literature have proved that they have substantial impacts on male fertility in vivo [ 8 , 10 , 33 ]. In fact, most of the lncRNAs’ function is still enigmatic.…”

Identification of male and female pupal characteristics of Zeugodacus cucurbitae (Coquillett) via machine vision

“…Therefore, the combination of the machine vision technology with the male sterility technology, to single out male pupae can substantially reduce the workload involved. Li et al used the RNAi technology to interfere with the lnC94638 gene expression in Z. cucurbitae (Coquillett) males to reduce their sperm number and fertility [26]. Moreover, they used the same technology to interfere with the ZcVMP26Ab gene in females to achieve low egg hatchability [27].…”