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Published on: January 7, 2019
Microarray-based approach identifies differentially expressed microRNAs in porcine sexually immature and mature
Lifan Luo1, Lianzhi Ye, Gang Liu
1Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, People's Republic of China.
Background:
MicroRNAs (miRNAs) are short non-coding RNA molecules which are proved to be involved in mammalian spermatogenesis. Their expression and function in the porcine germ cells are not fully understood.
Methodology:
We employed a miRNA microarray containing 1260 unique miRNA probes to evaluate the miRNA expression patterns between sexually immature (60-day) and mature (180-day) pig testes. One hundred and twenty nine miRNAs representing 164 reporter miRNAs were expressed differently (p<0.1). Fifty one miRNAs were significantly up-regulated and 78 miRNAs were down-regulated in mature testes. Nine of these differentially expressed miRNAs were validated using quantitative RT-PCR assay. Totally 15,919 putative miRNA-target sites were detected by using RNA22 method to align 445 NCBI pig cDNA sequences with these 129 differentially expressed miRNAs, and seven putative target genes involved in spermatogenesis including DAZL, RNF4 gene were simply confirmed by quantitative RT-PCR.
Conclusions:
Overall, the results of this study indicated specific miRNAs expression in porcine testes and suggested that miRNAs had a role in regulating spermatogenesis.
Insights
MicroRNAs (miRNAs) play a role in pig testes development. This study identified specific miRNAs and their potential targets, revealing their importance in regulating spermatogenesis.
Area of Science:
- Reproductive Biology
- Genetics
- Molecular Biology
Background:
- MicroRNAs (miRNAs) are crucial regulators in mammalian spermatogenesis.
- The specific roles and expression patterns of miRNAs in porcine germ cells remain largely uncharacterized.
Purpose of the Study:
- To investigate miRNA expression profiles in porcine testes at different developmental stages.
- To identify differentially expressed miRNAs and their potential target genes involved in spermatogenesis.
Main Methods:
- MicroRNA microarray analysis of testes from sexually immature (60-day) and mature (180-day) pigs.
- Validation of differentially expressed miRNAs using quantitative RT-PCR.
- Bioinformatic prediction of miRNA targets and confirmation of selected gene targets.
Main Results:
- A total of 129 miRNAs showed differential expression between immature and mature pig testes.
- 51 miRNAs were upregulated and 78 were downregulated in mature testes.
- Seven putative target genes, including DAZL and RNF4, were confirmed to be regulated by specific miRNAs.
Conclusions:
- Specific miRNA expression patterns are evident in porcine testes.
- These findings suggest a significant role for miRNAs in the regulation of spermatogenesis in pigs.
