Luo Xuliang, Guo Yan, Li Xuelian, Mei Zi, Zhou Haobo, Qiu Ping, Wang Haoxin, Chen Yan, Gong Yanzhang
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology and College of Veterinary Medicine, Huazhong Agricultural University, 430070, Wuhan, People's Republic of China.
BMC Genomics. 2025 Mar 28;26(1):305. doi: 10.1186/s12864-025-11500-5.
Aromatase, encoded by Cyp19a1, is the rate limiting enzyme in biosynthesis of estrogens, and excessive aromatase can reduce the semen quality in roosters. Seminal plasma extracellular vesicles (SPEV) are nanoscale vesicles that carry and transmit signaling molecules, thereby affecting semen quality. Currently it is still unclear whether SPEV are involved in the process of that aromatase affects the quality semen in chicken. To clarify this issue, lentivirus carrying Cyp19a1 (LV-CYP19A1) for over-expression of aromatase was constructed and injected to testis of 35-week-old roosters. Semen quality and seminal plasma hormone were measured, and SPEV were also extracted and proteome sequencing was performed after treatment of LV-CYP19A1.
The results indicated that semen volume, fertility, sperm motility, testosterone (T) levels were significantly decreased, and estradiol (E) levels were significantly increased in LV-CYP19A1 group than those in control group (P < 0.05). Through proteomic analysis of SPEV, we identified 966 differentially expressed proteins (DEPs) in the comparison of LV-CYP19A1 group and control group. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) items of DEPs are mainly enriched in ATP synthesis coupled electron transport, flagellated sperm motility, regulation of steroid biosynthetic process, and PI3K-Akt signaling pathway. Furthermore, 8 proteins including ENO4, APOB, SDHA, SDHB, UQCRC1, VIN, PITGB3 and FXN were identified as key proteins in SPEV involving in the process of aromatase regulated rooster semen quality.
Our results reveal that aromatase can down-regulate the protein expression related to regulation of ATP synthesis and metabolism, and sperm motility in SPEV, thereby reducing semen quality in roosters.
由Cyp19a1编码的芳香化酶是雌激素生物合成中的限速酶，过量的芳香化酶会降低公鸡的精液质量。精浆细胞外囊泡（SPEV）是携带和传递信号分子的纳米级囊泡，从而影响精液质量。目前尚不清楚SPEV是否参与芳香化酶影响鸡精液质量的过程。为了阐明这个问题，构建了携带Cyp19a1的慢病毒（LV-CYP19A1）以过表达芳香化酶，并将其注射到35周龄公鸡的睾丸中。测量精液质量和精浆激素，并在LV-CYP19A1处理后提取SPEV并进行蛋白质组测序。
结果表明，LV-CYP19A1组的精液量、受精率、精子活力、睾酮（T）水平显著降低，雌二醇（E）水平显著高于对照组（P <0.05）。通过对SPEV的蛋白质组分析，我们在LV-CYP19A1组和对照组的比较中鉴定出966种差异表达蛋白（DEP）。DEP的基因本体（GO）和京都基因与基因组百科全书（KEGG）条目主要富集在ATP合成偶联电子传递、鞭毛精子运动、类固醇生物合成过程的调节和PI3K-Akt信号通路中。此外，包括ENO4、APOB、SDHA、SDHB、UQCRC1、VIN、PITGB3和FXN在内的8种蛋白质被鉴定为SPEV中参与芳香化酶调节公鸡精液质量过程的关键蛋白质。
我们的结果表明，芳香化酶可以下调与ATP合成和代谢调节以及SPEV中精子活力相关的蛋白质表达，从而降低公鸡的精液质量。
