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microRNA profiling in three main stages during porcine spermatogenesis
Zonggang Luo1, Yingkai Liu, Lei Chen
1Department of Animal Science, Southwest University, Rongchang, Chongqing, China, luozgg@163.com.
Background:
Spermatogenesis is an intricate biological event wherein an undifferentiated spermatogonium develops into mature sperms. MicroRNAs are a type of single strand small non-coding RNA molecule and are implicated in the regulation of many crucial pathways during cell proliferation, apoptosis, and differentiation.
Method:
Here, we present a comprehensive comparison of miRNA expression profiling in three main stages during porcine spermatogenesis using high-throughput sequencing.
Results:
We built three small RNA libraries for the testis, the epididymis and the ejaculated sperm from a Landrace boar, and in total obtained 3821 precursor hairpins encoding for 4761 mature miRNAs, of which 23 are miRNA*. Notably, 940 precursor miRNAs produced both the 5'- and 3'- strands as sister pairs, indicating the distinctive expression patterns of germ cell miRNAs. Additionally, 418 out of 710 co-expressed miRNAs were identified as being differentially expressed between libraries (P < 0.001). Apart from the sexual specific X chromosome, many miRNAs were found to be located on chromosome 12, which may play potential roles in spermatogenesis according to the result of synteny analysis with human and mouse. The Gene Ontology and KEGG pathway analysis revealed that the target genes of co-expressed miRNAs were highly involved in the cell cycle process, metal ion binding, modification of plasma membrane, and the p53 signal pathway.
Insights
MicroRNAs regulate sperm development. This study profiled microRNA expression across porcine spermatogenesis stages, identifying key microRNAs and pathways involved in sperm formation.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Genomics
Background:
- Spermatogenesis is the complex process of sperm development from spermatogonia.
- MicroRNAs (miRNAs) are small non-coding RNAs regulating gene expression in cell proliferation, apoptosis, and differentiation.
Purpose of the Study:
- To comprehensively compare miRNA expression profiles during three key stages of porcine spermatogenesis.
- To identify specific miRNAs and their target pathways involved in sperm development.
Main Methods:
- High-throughput sequencing was used for miRNA expression profiling.
- Small RNA libraries were constructed from porcine testis, epididymis, and ejaculated sperm.
- Bioinformatic analyses included differential expression, synteny, Gene Ontology, and KEGG pathway analysis.
Main Results:
- Over 4700 mature miRNAs were identified, with distinct expression patterns observed.
- 418 co-expressed miRNAs showed differential expression between the analyzed tissues (P < 0.001).
- Target genes of co-expressed miRNAs are enriched in cell cycle, metal ion binding, plasma membrane modification, and the p53 signaling pathway.
Conclusions:
- Distinct miRNA expression profiles are associated with different stages of porcine spermatogenesis.
- Specific miRNAs and their target pathways, including the p53 pathway, are crucial for germ cell development.
- Chromosome 12 and the X chromosome harbor miRNAs potentially important for spermatogenesis.
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