Abstract
Cattleyak are interspecific hybrids between cattle and yak, exhibiting the same prominent adaptability as yak and much higher performances than yak. However, male infertility of cattleyak resulted from spermatogenic arrest has greatly restricted their effective utilization in yak breeding. In past decades, much work has been done to investigate the mechanisms of spermatogenic arrest, but little is known about the differences of the post-transcriptional regulators between cattleyak and yak, which may contribute to the impaired spermatogenesis. MiRNAs, a class of endogenous non-coding small RNA, were revealed to play crucial roles in regulating gene expression at post-transcriptional level. In the present study, we identified 50 differentially expressed (DE) known miRNAs and 11 novel miRNAs by using Illumina HISeq and bioinformatic analysis. A total of 50 putative target sites for the 13 DE known miRNAs and 30 for the 6 DE novel miRNAs were identified, respectively. GO and KEGG analyses were performed to reveal the functions of target genes for DE miRNAs. In addition, RT-qPCR was performed to validate the expression of the DE miRNAs and its targets. The identification of these miRNAs may provide valuable information for a better understanding of spermatogenic arrest in cattleyak.
Highlights
Cattleyak are interspecific hybrids between cattle (♂) and yak (♀), which exhibit the same prominent adaptability to harsh environment on Qinghai-Tibetan Plateau as yak
In order to identify differentially expressed (DE) miRNAs involved in spermatogenic arrest of cattleyak, small RNA libraries were constructed from testis total RNAs of cattleyak and yak individuals for Illumina HISeq and miRNA screening
The numbers of small RNAs obtained from the six samples ranged from 4920921 to 11294549, with the annotated percentages ranging from 0.2% to 0.8% and ambiguously annotated percentages ranging from 0.1% to 0.2% (Table 1)
Summary
Cattleyak are interspecific hybrids between cattle (♂) and yak (♀), which exhibit the same prominent adaptability to harsh environment on Qinghai-Tibetan Plateau as yak. Spermatogenesis is a highly-regulated process comprising three main continuous stages of germ cell differentiation: mitotic proliferation of spermatogonia, meiosis of spermatocytes and spermiogenesis of haploid spermatids. As the emerging roles of miRNAs discovered in regulating gene expression at the post-transcriptional level, investigation of the regulatory functions of miRNAs involved in spermatogenesis will contribute to a better understanding of the mechanisms of spermatogenic arrest of cattleyak. We identified the DE miRNAs between the testis samples of cattleyak and yak, some of which were involved in gene regulation during mitotic proliferation, meiosis and spermiogenesis processes. The further characterization of these miRNAs is helpful to reveal the mechanisms of spermatogenic arrest of cattleyak and the identified miRNAs together with their target genes may serve as effective molecular markers in resolving the problems of male infertility of cattleyak in the future
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