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https://doi.org/10.5713/ab.21.0259    [Accepted] Published online August 25, 2021.
Alterations of mRNA and lncRNA profiles associated with the extracellular matrix and spermatogenesis in goats
Haolin Chen1,2  , Xiaomeng Miao2  , Jinge Xu2  , Ling Pu2  , Liang Li2  , Yong Han2  , Fengxian Mao3  , Youji Ma1,* 
1College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, 730000, China
2Institute of Animal Husbandry and Veterinary, Guizhou Academy of Agricultural Sciences, Guizhou, 550000, China
3Guizhou Province Livestock and Poultry Genetic Resources Management Station, Guiyang, Guizhou, 550000, China
Correspondence:  Youji Ma,Email: yjma@gsau.edu.cn
Received: 3 June 2021   • Revised: 12 July 2021   • Accepted: 16 August 2021
Abstract
Objective
Spermatozoa are produced within the seminiferous tubules after sexual maturity. The expression levels of mRNAs and lncRNAs in testicular tissues are different at each stage of testicular development and are closely related to formation of the extracellular matrix and spermatogenesis. Therefore, we set out to study the expression of lncRNAs and mRNAs during the different developmental stages of the goat testis.
Methods
We constructed 12 RNA libraries using testicular tissues from goats aged 3, 6, and 12 months, and studied the functions of mRNAs and lncRNAs using the Gene Ontogeny (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Relationships between differentially expressed genes (DEGs) were analyzed by lncRNA-mRNA co-expression network and protein-protein interaction network (PPI). Finally, the protein expression levels of matrix metalloproteinase 2 (MMP2), insulin-like growth factor 2 (IGF2), and insulin-like growth factor-binding protein 6 (IGFBP6) were detected by western blotting.
Results
We found 23, 8, and 135 differentially expressed lncRNAs and 161, 12, and 665 differentially expressed mRNAs that were identified between 3 vs. 6, 6 vs. 12, and 3 vs. 12 months, respectively. GO, KEGG, and PPI analyses showed that the differential genes were mainly related to the extracellular matrix. Moreover, MMP2 was a hub gene and co-expressed with the lncRNA TCONS-0002139 and TCONS-00093342. The results of qRT-PCR verification were consistent with those of RNA-seq sequencing. The expression trends of MMP2, IGF2, and IGFBP6 protein were the same as that of mRNA, which all decreased with age. IGF2 and MMP2 were significantly different in the 3 vs. 6-month-old group (P < 0.05).

Conclusion

These results improve our understanding of the molecular mechanisms involved in sexual maturation of the goat testis.
Keywords: Goat; LncRNA; mRNA; Puberty; Testis
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