piRNA‐19128 regulates spermatogenesis by silencing of KIT in chicken

Abstract

Spermatogenesis is a complex process. Some studies have shown that Piwi-interacting RNAs (piRNAs) play an important role in spermatogenesis. To verify the evaluate between piRNAs and PIWI proteins in chicken and its possible role in spermatogenesis and reproductive stem cell proliferation and differentiation, we performed immunoprecipitation and deep sequencing analyses and determined the expression profiles of small RNAs in primordial germ cells (PGCs), spermatogonial stem cells (SSCs), spermatogonia (Sa) cells, and spermatozoa. Length analysis showed that piRNAs bound to PIWIL1 mainly contained 23-30 nt. Base preference analysis showed "1U-10A"; moreover, base preference of piRNAs was obvious in all of germline cells. Here we reported the TE family of gallus gallus, and targeted by piRNA. Target gene of piRNA annotation enrichment analysis identified candidate genes KIT, SRC, WNT4, and HMGB2. Kyoto Encyclopedia of Genes and Genomes analysis showed that these genes were associated with steroid hormone biosynthesis, Notch signaling pathway, and melanogenesis. These results indicate that chicken piRNAs perform important regulatory roles during spermatogenesis similar to mice piRNAs. Chicken piRNAs interacted with PIWI proteins and regulated spermatogenesis and germ cell proliferation and differentiation. Further, we observed a negative correlation between piRNA-19128 and KIT expression. Results of dual-luciferase reporter assay confirmed that piRNA-19128 directly interacted with KIT, suggesting that it plays a key role in the regulation spermatogenesis by inhibiting KIT expression. Thus, the present study provides information on the length and base preference of chicken piRNAs and suggests that piRNA-19128 regulates spermatogenesis in chicken by silencing KIT.

The gene regulatory role of non-coding RNAs in non-obstructive azoospermia

“…PiRNA is mainly expressed in germ cells, and many studies have confirmed that piRNA is related to spermatogenesis, loss of sperm motility and abnormal sperm morphology (96)(97)(98). Therefore, piRNA is an excellent study object in the area of NOA.…”

miR-301a-5p Regulates TGFB2 during Chicken Spermatogenesis

“…The miRNA expression data used in the present study were obtained from our previous study [11,15]. The data were analyzed for differences in miRNA expression levels during spermatogenesis.…”

“…Recently, many studies have shown that several regulator types which include methylation, ubiquitination, acetylation, transcription factor and et.al., play major roles in germline cell development and spermatogenesis [7][8][9][10][11][12]. Small RNAs, which are a class of short non-coding RNAs, play a vital role during the entire spermatogenesis process [13][14][15].…”

PAZ Domain is Critical for PIWIL1 Function During Spermatogenesis in Chicken

“…Therefore, spermatogenesis depends on the stage-speci c expression of several genes encoding regulatory factors, including P element-induced wimpy testis (Piwi)1 [2]. Piwi was rst discovered in Drosophila, where it was shown to regulate asymmetric division of germline stem cells [3,4]. Later, it has been established that Piwi proteins are highly conserved among plants and animals.…”

“…The association between Piwi proteins and piRNAs is known to suppress transposable elements and protect the integrity of the genome in germ cells [10]. We and other researchers have recently shown that PIWIL1 plays an important role in spermatogenesis [4]. Even though many studies have shown that each domain of Piwi protein plays an important role in the process of germ cell development.…”