Seminal Plasma Exosomes: Promising Biomarkers for Identification of Male and Pseudo-Males in Cynoglossus semilaevis
In mammals, small RNAs enclosed in exosomes have been identified as appropriate signatures for disease diagnosis. However, there is limited information on exosomes derived from seminal plasma, and few studies have reported analyzed the composition of exosomes and enclosed small RNAs in fish. The half-smooth tongue sole (Cynoglossus semilaevis) is an economically important fish for aquaculture, and it exhibits sexual dimorphism: the female gender show higher growth rates and larger body sizes than males. Standard karyotype analysis and artificial gynogenesis tests have revealed that this species uses heterogametic sex determination (ZW/ZZ), and so-called sex-reversed pseudo-males exist. In this study, we successfully identified exosomes in the seminal plasma of C. semilaevis; to the best of our knowledge, this is the first report of exosomes in fish seminal plasma. Analysis of the nucleotide composition showed that miRNAs were dominant in the exosomes, and the miRNAs were sequenced and compared to identify signature miRNAs as sexual biomarkers. Moreover, target genes of the signature miRNAs were predicted by sequence matching and annotation. Finally, four miRNAs (dre-miR-141-3P, dre-miR-10d-5p, ssa-miR-27b-3p, and ssa-miR-23a-3p) with significant differential expression in the males and pseudo-males were selected from the signature candidate miRNAs as markers for sex identification, and their expression profiles were verified using real-time quantitative PCR. Our findings could provide an effective detection method for sex differentiation in fish.
KeywordsExosomes Seminal plasma MicroRNAs Sexual biomarker Cynoglossus semilaevis
We thank Dr. Zhang Fan of OE Biotech Ltd. (Shanghai, China) for her critique of this manuscript. This work was supported by grants from the Tianjin Natural Science Foundation (17JCQNJC15000), transformation project of Tianjin Agricultural Achievements (201604090), special funding for Modern Agricultural Industrial Technology System (CARS-47-Z01), Modern Industrial Technology System in Tianjin (ITTFRS2017011), and National Natural Science Foundation of China (31872546, 31472262), the China-ASEAN Maritime Cooperation Fund through the project “China-ASEAN Center for Joint Research and Promotion of Marine Aquaculture Technology.”
Bo Zhang, Baolong Bao, and Jinsheng Sun conceived and designed the study. Bo Zhang and Na Zhao performed the computational analysis and expression profiling. Lei Jia and Na Zhao performed the identification of exosome experiments. Bo Zhang and Na Zhao wrote the manuscript. All authors read and approved the final manuscript. Na Zhao and Kunming Li performed to the RT-qPCR analysis.
Compliance with Ethical Standards
All experiments were approved by the Animal Care Committee of Shanghai Ocean University.
Conflict of Interest
The authors declare that they have no conflict of interest.
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