1700108J01Rik and 1700101O22Rik are mouse testis-specific long non-coding RNAs

Abstract

Long non-coding RNAs (lncRNAs; > 200 nucleotides in length) have attracted attention as fine-tuners of gene expression. However, little is known about the cell- and stage-specific expression pattern and function of lncRNAs in spermatogenesis. The purpose of this study was to identify mouse testis-associated lncRNAs using a combination of computational and experimental approaches. We first used the FANTOM5 database to survey lncRNA expression in the mouse testis and performed reverse transcription quantitative polymerase chain reaction (real-time PCR) and in situ hybridization (ISH) analyses. In silico analysis showed that most of the highly expressed lncRNAs in the adult mouse testis were testis-specific lncRNAs and were expressed at and following the initiation of spermatogenesis. We selected the antisense lncRNA 1700108J01Rik and long intergenic non-coding RNA 1700101O22Rik from the most highly expressed lncRNAs in the adult testis for further analysis. Real-time PCR analysis confirmed that 1700108J01Rik and 1700101O22Rik were specifically expressed in the testis. ISH analysis revealed that the two mouse-testis-specific lncRNAs were expressed exclusively in testicular germ cells in meiotic prophase and the round spermatid stage, which coincide with the period of transcriptional reactivation during spermatogenesis. The cytoplasmic distribution of these lncRNAs revealed by ISH suggests their involvement in post-transcriptional gene regulation rather than in epigenetic or transcriptional regulation. Our data provide new insight into testis-associated lncRNAs that will be useful in expression and functional studies of spermatogenesis.

Long noncoding RNAs: new insights in modulating mammalian spermatogenesis

“…Furthermore, correlation analysis between mRNAs and lncRNAs showed high correlation coefficients and suggested coordinated changes in their expression during the biogenesis and function of male spermatogenesis. A similar investigation reported the expression of antisense lncRNA 1700108J01Rik and long intergenic noncoding RNA 1700101O22Rik [53] in meiotic prophase and round spermatid stages in testicular germ cells. Similarly, Dai et al reported higher expression of lncRNA-testicular cell adhesion molecule 1 (lncRNA-Tcam1) in round spermatids than in somatic cells or germ cells in mice, with lncRNA-Tcam1 being localized in 45% of the cells found in testis [55].…”

“…The ChIPBase, NPInter, miRcode, DIANA-LncBase, StarBasev2.0, lncRNA2Target, and lncRNADisease are examples of interaction databases. The specific databases include lnCeDB, NRED, Linc2go, and lncRNASNP [49][50][51][52][53][54][55][56][57][58][59].…”

“…Sun et al conducted a microarray analysis to assess the lncRNA expression profiles during postnatal development in mouse testis [21]. Song et al explored the testis-specific lncRNAs and their expression in adult mouse testis using experimental and computational methods [53] and reported highly expressed lncRNAs in adult testis. Wichman et al analyzed the dynamic expression pattern of lncRNAs in the testis that escapes the meiotic sex chromosome inactivation (MSCI) [54].…”

“…Other lncRNAs are via an in cis-mediated mechanism to regulate neighboring gene expression [85,86]. Song et al reported the cytoplasmic distribution of testis-specific lncRNAs involved in post-transcriptional gene regulation [53].…”

“…Enhancer-associated lncRNAs participate in transcriptional activation by acting over long distances on distal promoters associated with protein factors and the modulation of chromatin structures [87]. Song et al revealed the cytoplasmic distribution of 1700108J01Rik and 1700101O22Rik using in situ hybridization, suggesting their function in post- Highly expressed in pachytene spermatocytes [54] Tesra 4435 bp Highly expressed in the nuclei of pachytene spermatocytes [82] LncRNA-Tcam1 2.4 kb Expressed in mouse male germ cells [55] LncRNA033862 ∼6.4 kb Highly expressed in mouse spermat-ogonial stem cells especially in early spermatogonia [80] a The chromosomal location is in the mouse genome transcriptional gene regulation mainly in the nucleus; however, they are not involved in epigenetic and transcriptional regulation [53]. Similarly, Zhang et al reported the post-transcriptional role of lncRNA Dmrt1 on chromosome 5 in spermatogenesis and testis development in mice [65].…”