Integrated Analysis of miRNA and mRNA Expression Profiles Reveals Functional miRNA-Targets in Development Testes of Small Tail Han Sheep

Abstract

Small Tail Han Sheep is a highly valued local breed in China because of their precocity, perennial estrus, and high fecundity. The average annual lambing rate of ewes is as high as 180–270%, the semen of ram has characteristics of high yield, high density, and good motility. To reveal the key miRNAs and miRNA-targets underlying testis development and spermatogenesis in male Small Tail Han Sheep, integrated analysis of miRNA and mRNA expression profiles in 2-, 6-, and 12-month-old testes was performed by RNA-seq technology and bioinformatics methods. The results showed that total of 153 known sheep miRNAs and 2712 novel miRNAs were obtained in 2-,6 - and 12-month-old Small Tail Han Sheep testes; 5, 1, and 4 differentially expressed (DE) known sheep miRNAs, and 132, 105, and 24 DE novel miRNAs were identified in 2- vs. 6-, 6- vs. 12-, and 2- vs. 12-month-old testes, respectively. We combined miRNA results of this study and the mRNA results obtained in our previous study to predict the target mRNAs of DE known sheep miRNAs; 131, 10, and 15 target mRNAs of DE known sheep miRNAs and 76, 1, and 11 DE miRNA–targets were identified in the three groups, respectively. GO and KEGG analyses showed that: in 2- vs. 6-month-olds, the target genes of DE known sheep miRNAs were involved in 100 biological processes and 11 signaling pathways; in 6- vs. 12-month-olds, the target genes of DE known sheep miRNAs were involved in 4 biological processes; and in 2- vs. 12-month-olds, the target genes of DE known sheep miRNAs were involved in 17 biological processes and 4 signaling pathways. Three miR–target regulatory networks were constructed based on these DE miRNA–targets. The key miRNA-Targets involved in testis development and spermatogenesis were screened. 6 known sheep miRNAs and 6 novel miRNAs were selected to validate the accuracy of miRNA sequencing data by qRT-PCR. The binding sites of oar-miR-379-5p with WNT8A was validated by a dual luciferase reporter gene detection system.

Expression Analysis of Circular RNAs in Young and Sexually Mature Boar Testes

“…For example, a large number of circRNAs were identified in the testicular tissue or seminal plasma of Qinchuan cattle [ 24 ], rats [ 28 ], humans [ 25 , 42 ], and pigs [ 26 ]. Related research on pigs has been mainly focused on maps of the testicular transcriptome [ 43 ], miRNAs [ 44 ], and sperm [ 45 ]. However, the comparative expression and the potential function of circRNA in development of pig testis have not been hitherto investigated.…”

MicroRNA Expression Profile in Peripheral Blood Lymphocytes of Sheep Vaccinated with Nigeria 75/1 Peste Des Petits Ruminants Virus

“…The miR-17 has been reported to bind the BVDV genome leading to changes in the host transcriptome [61]. The oar-miR-379-5p and oar-miR-30a might play key roles in apoptosis [62] and PPRV was also reported to cause apoptosis of host cells [63].…”

Integrating miRNA and mRNA Profiling to Assess the Potential miRNA–mRNA Modules Linked With Testicular Immune Homeostasis in Sheep

“…RNA sequencing (mRNA-seq and sRNA-seq) has emerged as a powerful tool to identify and characterize the genes and miRNAs expressed in mammalian testes ( 18 , 19 ). RNA sequencing is of great significance for further filtering the immune-related mRNAs–miRNAs in testes and understanding the complex processes that modulate the maintenance of the testes' immune privilege.…”