Journal of Developmental Medicine(Electronic Version)  2023, Vol. 11 Issue (2): 81-91 DOI: 10.3969/j.issn.2095-5340.2023.02.001 |
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| Differential expression analysis and related pathways of testis based on transcriptomesequencing in Ppp2ca conditional knockout mice |
| Liu Xing, Chen Bingyan, Wang Danni,et al
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Reproductive Medical Center, theAffiliated Changzhou No.2 People’s Hospital of Nanjing Medical Unversity, Jiangsu, Changzhou 213000,China
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Abstract 【Abstract】 Objective To investigate the effect of conditional knockout of protein phosphatase 2 catalyticsubunit alpha (Ppp2ca) on differentially expressed genes (DEGs) and related pathways in mouse testiculargerm cells. Method Mice with conditional knockout Ppp2ca (Ppp2cacKO) in germ cells were constructedby Cre-loxP principle. The role of protein phosphatase 2A (PP2A) in the male reproductive system wasexplored by inactivating the catalytic subunit of PP2A. When the knockout male mice were 8 weeks old,the body weight and testis weight were recorded, the testis and epididymis of the mice were collected forhistological examination, and the serum testosterone level was detected. Testicular tissues were collectedfor transcriptome sequencing, gene ontology analysis (GO) and Kyoto encyclopedia of genes and genomes(KEGG) bioinformatics analysis. Result The testis weight of Ppp2cacKO mice and Ppp2cactrl mice were(0.044±0.003) and (0.119±0.008) g (t=24.550, P<0.001). The serum testosterone level was (0.327±0.096)and (0.567±0.050) μg/L (t=3.899, P=0.018). The testis weight and serum testosterone level of Ppp2cacKOmice were lower than those of Ppp2cactrl mice. Hematoxylin-eosin staining showed that no sperm wasobserved in the testis and epididymis of the Ppp2cacKO mice, and the number and hierarchy of spermatogeniccells in seminiferous tubules were significantly reduced with a large number of vacuolar structures. A totalof 11 304 DEGs (7 568 up-regulated and 3 736 down-regulated), 36 microRNAs (8 up-regulated and 28down-regulated), and 3 732 long non-coding RNAs (1 107 up-regulated and 2 625 down-regulated) werescreened by transcriptome sequencing. The DEGs in Ppp2cacKO mice were significantly different in multipleGO enrichment terms, such as spermatogenesis, spermatid development, sperm motility, acrosomal vesicles,centrosomes, microtubules and mitochondrial outer membrane. In addition, KEGG functional enrichmentresults showed that DEGs were mainly related to biochemical metabolic pathways and signal transductionpathways, and mainly affected endocytosis, actin cytoskeleton regulation, cAMP signaling pathway, Wntsignaling pathway, lipid metabolism, cytochromic P450 and sphingomodin signaling pathway, etc. TheseDEGs closely related with spermatogenesis. Conclusion Knockout of Ppp2ca can cause abnormalfunction of PP2A, affecting spermatogenesis and the activity of signaling pathways related to testiculardevelopment, leading to impaired spermatogenesis, meiotic arrest and low testosterone level in mice.
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Received: 10 January 2023
Published: 31 March 2023
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