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Screening of Biomarkers Related to Ovarian Maturation and Spawning in Blunt Snout Bream (Megalobrama amblycephala) Based on Metabolomics and Transcriptomics

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Abstract

In fish breeding practices, gamete maturity of females is vital to reproductive success. For some species, it is possible to estimate the female maturation status based on abdomen observation, but quite difficult for some species which mature at big size. To screen out the potential biomarker in fish blood relating to female maturation, we employed the approach integrating the UPLC-MS/MS and RNA-seq techniques to investigate the metabolites and genes reflecting the sexual maturation and spawning of female blunt snout bream Megalobrama amblycephala. The study included four groups, 1-year-old immature female individuals, 2-year-old immature female individuals, 2-year-old sexually mature female individuals, and 2-year-old sexually mature female individuals after 24 h of successful spawning. The upregulated metabolites in mature females were involved in “steroid hormone biosynthesis,” “metabolic pathways,” “glycerophospholipid metabolism,” etc. compared with those of immature individuals. As the key intermediate of steroid hormone biosynthesis, 17α-hydroxypregnenolone exhibited the highest level in 2-year-old mature females than in the immature females. Meanwhile, the metabolites (i.e., dodecanoic acid and myristic acid) participating in fatty acid synthesis exhibited much lower levels in the females after spawning than those before spawning. In addition to the metabolites, the genes involved in ovarian steroidogenesis were significantly upregulated in the 2-year-old immature females compared to the 1-year-old immature females, indicating that the ovarian steroidogenesis plays important roles in ovarian development of M. amblycephala at the early stages. The significant upregulation of genes (i.e., itpr1, camk2, and mekk2) involved in the “GnRH signaling pathway” was observed in the mature females compared with the immature females, which indicated that the estrogen levels increased after female maturation in M. amblycephala. Moreover, many genes (e.g., gck, creb1, tf2-9, ryr2, asgr1, and creb1) regulating insulin secretion and thyroid hormone synthesis were significantly downregulated after female spawning. The dynamics of gene expression and metabolites observed in this study provide novel cues for guiding fish practical artificial reproduction.

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Data Availability

The raw data of metabolites was deposited at EMBL-EBI MetaboLights under accession number MTBLS1020. Gene expression profiles were deposited at the Gene Expression Omnibus database (GEO) under accession number GSE131093. The raw data of RNA-Seq used as reference was deposited at the NCBI Sequence Read Archive (SRA) (NCBI Sequence Read Archive SRX5824305).

References

  • Bromage NR, Whitehead C, Breton B (1982) Relationships between serum levels of gonadotropin, oestradiol-17β, and vitellogenin in the control of ovarian development in the rainbow trout: II. The effects of alterations in environmental photoperiod. Gen Comp Endocrinol 47:366–376

    Article  CAS  PubMed  Google Scholar 

  • Budzinski H, Devier MH, Labadie P, Togola A (2006) Analysis of hormonal steroids in fish plasma and bile by coupling solid-phase extraction to GC/MS. Anal Bioanal Chem 386:1429–1439

    Article  CAS  PubMed  Google Scholar 

  • Chen S, Zhou Y, Chen Y, Gu J (2018) Fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 34:i884–i890

  • Devlin RH, Nagahama Y (2002) Sex determination and sex differentiation in fish: an overview of genetic, physiological, and environmental influences. Aquaculture 208:191–364

    Article  CAS  Google Scholar 

  • Dufour S, Lopez E, Le Menn F, Le Belle N, Baloche S, Fontaine YA (1988) Stimulation of gonadotropin release and of ovarian development, by the administration of a gonadoliberin agonist and of dopamine antagonists, in female silver eel pretreated with estradiol. Gen Comp Endocrinol 70:20–30

    Article  CAS  PubMed  Google Scholar 

  • Filby AL, Thorpe KL, Maack G, Tyler CR (2007) Gene expression profiles revealing the mechanisms of anti-androgen-and estrogen-induced feminization in fish. Aquat Toxicol 81:219–231

    Article  CAS  PubMed  Google Scholar 

  • Gao Z, Luo W, Liu H, Zeng C, Liu X, Yi S, Wang W (2012) Transcriptome analysis and SSR/SNP markers information of the blunt snout bream (Megalobrama amblycephala). PLoS One 7:e42637

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gil-Solsona R, Nácher-Mestre J, Lacalle-Bergeron L, Sancho JV, Calduch-Giner JA, Hernández F, Pérez-Sánchez J (2017) Untargeted metabolomics approach for unraveling robust biomarkers of nutritional status in fasted gilthead sea bream (Sparus aurata). PeerJ 5:e2920

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Good C, Davidson J (2016) A review of factors influencing maturation of Atlantic salmon, Salmo salar, with focus on water recirculation aquaculture system environments. J World Aquacult Soc 47:605–632

    Article  Google Scholar 

  • Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29:644

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guan N, Zhou Q, Lan T, Zhou L, Zhao B, Wang W, Gao Z (2018) Is the nutritional value of fish fillet related to fish maturation or fish age? Integrated analysis of transcriptomics and metabolomics in blunt snout bream (Megalobrama amblycephala). Cell Physiol Biochem 49:17–39

    Article  CAS  PubMed  Google Scholar 

  • Guo C, Huang X, Yang M, Wang S, Ren S, Li H, Peng X (2014) GC/MS-based metabolomics approach to identify biomarkers differentiating survivals from death in crucian carps infected by Edwardsiella tarda. Fish Shellfish Immunol 39:215–222

    Article  CAS  PubMed  Google Scholar 

  • Gutierrez AP, Lubieniecki KP, Fukui S, Withler RE, Swift B, Davidson WS (2014) Detection of quantitative trait loci (QTL) related to grilsing and late sexual maturation in Atlantic salmon (Salmo salar). Mar Biotechnol (NY) 16:103–110

    Article  CAS  Google Scholar 

  • Haidle L, Janssen JE, Gharbi K, Moghadam HK, Ferguson MM, Danzmann RG (2008) Determination of quantitative trait loci (QTL) for early maturation in rainbow trout (Oncorhynchus mykiss). Mar Biotechnol (NY) 10:579–592

    Article  CAS  Google Scholar 

  • Hamasaki M, Takeuchi Y, Miyaki K, Yoshizaki G (2013) Gonadal development and fertility of triploid grass puffer Takifugu niphobles induced by cold shock treatment. Mar Biotechnol (NY) 15:133–144

    Article  CAS  Google Scholar 

  • Irvine I, Sorensen PW (1993) Acute olfactory sensitivity of wild common carp, Cyprinus carpio, to goldfish hormonal sex pheromones is influenced by gonadal maturity. Can J Zool 71:2199–2210

    Article  CAS  Google Scholar 

  • Knight OM, Van Der Kraak G (2015) The role of eicosanoids in 17α, 20β-dihydroxy-4-pregnen-3-one-induced ovulation and spawning in Danio rerio. Gen Comp Endocrinol 213:50–58

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi M, Sorensen PW, Stacey NE (2002) Hormonal and pheromonal control of spawning behavior in the goldfish. Fish Physiol Biochem 26:71–84

    Article  CAS  Google Scholar 

  • Kocmarek AL, Ferguson MM, Danzmann RG (2015) Comparison of growth-related traits and gene expression profiles between the offspring of neomale (XX) and normal male (XY) rainbow trout. Mar Biotechnol (NY) 17:229–243

    Article  CAS  Google Scholar 

  • Kuo C, Nash CE (1975) Recent progress on the control of ovarian development and induced spawning of the grey mullet (Mugil cephalus L.). Aquaculture 5:19–29

    Article  Google Scholar 

  • Langmead B, Salzberg SL (2012) Fast gapped-read alignment with bowtie 2. Nat Methods 9:357

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Luo W, Zeng C, Deng W, Robinson N, Wang W, Gao Z (2014) Genetic parameter estimates for growth-related traits of blunt snout bream (Megalobrama amblycephala) using microsatellite-based pedigree. Aquac Res 45:1881–1888

    Article  Google Scholar 

  • Melamed P, Rosenfeld H, Elizur A, Yaron Z (1998) Endocrine regulation of gonadotropin and growth hormone gene transcription in fish. Comp Biochem Physiol C 119:325–338

    Article  CAS  PubMed  Google Scholar 

  • Nagahama Y, Yamashita M (2008) Regulation of oocyte maturation in fish. Develop Growth Differ 50:S195–S219

    Article  CAS  Google Scholar 

  • Nishimura T, Tanaka M (2014) Gonadal development in fish. Sex Dev 8:252–261

    Article  PubMed  Google Scholar 

  • Norberg B, Weltzien F, Karlsen Ø, Holm JC (2001) Effects of photoperiod on sexual maturation and somatic growth in male Atlantic halibut (Hippoglossus hippoglossus L.). Comp Biochem Physiol B 129:357–365

    Article  CAS  PubMed  Google Scholar 

  • Patiño R, Sullivan CV (2002) Ovarian follicle growth, maturation, and ovulation in teleost fish. Fish Physiol Biochem 26:57–70

    Article  Google Scholar 

  • Pavlidis M, Greenwood L, Mourot B, Kokkari C, Le Menn F, Divanach P, Scott AP (2000) Seasonal variations and maturity stages in relation to differences in serum levels of gonadal steroids, vitellogenin, and thyroid hormones in the common dentex (Dentex dentex). Gen Comp Endocrinol 118:14–25

    Article  CAS  PubMed  Google Scholar 

  • Picone G, Balling Engelsen S, Savorani F, Testi S, Badiani A, Capozzi F (2011) Metabolomics as a powerful tool for molecular quality assessment of the fish Sparus aurata. Nutrients 3:212–227

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Power DM, Llewellyn L, Faustino M, Nowell MA, Björnsson BT, Einarsdóttir IE, Canario AV, Sweeney GE (2001) Thyroid hormones in growth and development of fish. Comp Biochem Physiol C 130:447–459

    CAS  Google Scholar 

  • Prisingkorn W, Prathomya P, Jakovlić I, Liu H, Zhao Y, Wang W (2017) Transcriptomics, metabolomics and histology indicate that high-carbohydrate diet negatively affects the liver health of blunt snout bream (Megalobrama amblycephala). BMC Genomics 18:856

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Qiao Q, Le Manach S, Sotton B, Huet H, Duvernois-Berthet E, Paris A, Duval C, Ponger L, Marie A, Blond A, Mathéron L, Vinh J, Bolbach G, Djediat C, Bernard C, Edery M, Marie B (2016) Deep sexual dimorphism in adult medaka fish liver highlighted by multi-omic approach. Sci Rep-UK 6:32459

    Article  CAS  Google Scholar 

  • Roufidou C, Schmitz M, Mayer I, Sebire M, Katsiadaki I, Shao YT, Borg B (2018) Hormonal changes over the spawning cycle in the female three-spined stickleback, Gasterosteus aculeatus. Gen Comp Endocrinol 257:97–105

    Article  CAS  PubMed  Google Scholar 

  • Senthilkumaran B, Yoshikuni M, Nagahama Y (2004) A shift in steroidogenesis occurring in ovarian follicles prior to oocyte maturation. Mol Cell Endocrinol 215:11–18

    Article  CAS  PubMed  Google Scholar 

  • Škugor A, Slanchev K, Torgersen JS, Tveiten H, Andersen Ø (2014) Conserved mechanisms for germ cell-specific localization of nanos3 transcripts in teleost species with aquaculture significance. Mar Biotechnol (NY) 16(3):256–264

    Article  CAS  Google Scholar 

  • Sower SA, Schreck CB (1982) Steroid and thyroid hormones during sexual maturation of coho salmon (Oncorhynchus kisutch) in seawater or fresh water. Gen Comp Endocrinol 47:42–53

    Article  CAS  PubMed  Google Scholar 

  • Stacey NE, MacKenzie DS, Marchant TA, Kyle L, Peter RE (1984) Endocrine changes during natural spawning in the white sucker, Catostomus commersoni: I. gonadotropin, growth hormone, and thyroid hormones. Gen Comp Endocrinol 56:333–348

    Article  CAS  PubMed  Google Scholar 

  • Tang W, Gilman AG (1992) Adenylyl cyclases. Cell 70:869–872

    Article  CAS  PubMed  Google Scholar 

  • Tarazona S, García F, Ferrer A, Dopazo J, Conesa A (2011) NOIseq: a RNA-seq differential expression method robust for sequencing depth biases. EMBnet J 17:18–19

    Article  Google Scholar 

  • Treerattrakool S, Panyim S, Udomkit A (2011) Induction of ovarian maturation and spawning in Penaeus monodon broodstock by double-stranded RNA. Mar Biotechnol (NY) 13:163–169

    Article  CAS  Google Scholar 

  • Trudeau VL (1997) Neuroendocrine regulation of gonadotrophin II release and gonadal growth in the goldfish, Carassius auratus. Rev Reprod 2:55–68

    Article  CAS  PubMed  Google Scholar 

  • Waiho K, Fazhan H, Zhang Y, Zhang Y, Li S, Zheng H, Liu W, Ikhwanuddin M, Ma H (2019) Gonadal microRNA expression profiles and their potential role in sex differentiation and gonadal maturation of mud crab Scylla paramamosain. Mar Biotechnol (NY) 21:320–334

    Article  CAS  Google Scholar 

  • Wen B, Mei Z, Zeng C, Liu S (2017) metaX: a flexible and comprehensive software for processing metabolomics data. BMC Bioinformatics 18:183

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xie C, Mao X, Huang J, Ding Y, Wu J, Dong S, Kong L, Gao G, Li C, Wei L (2011) KOBAS 2.0: a web server for annotation and identification of enriched pathways and diseases. Nucleic Acids Res 39:W316–W322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yaron Z, Levavi-Sivan B (2011) Endocrine regulation of fish reproduction. Encyclopedia of fish physiology: from genome to environment. Elsevier, pp. 1500-1508

  • Zhou L, Zhao B, Guan N, Wang W, Gao Z (2017) Plasma metabolomics profiling for fish maturation in blunt snout bream. Metabolomics 13:40

    Article  CAS  Google Scholar 

  • Zohar Y, Mylonas CC (2001) Endocrine manipulations of spawning in cultured fish: from hormones to genes, reproductive biotechnology in finfish aquaculture. Elsevier, pp. 99–136

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Funding

This work was financially supported by the National Natural Science Foundation of China (Grant No. 31872559), National Key Research and Development Program (Grant No. 2018YFD0900205), Modern Agriculture Industry Technology System Construction Projects of China titled as-Staple Freshwater Fishes Industry Technology System (Grant No. CARS-46-08), and Fundamental Research Funds for the Central Universities (Grant No. 2662018PY035).

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  1. Shaokui Yi and Li-Fang Liu contributed equally to this work.

Authors and Affiliations

  1. College of Fisheries, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education/Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, No. 1 Shizishan Street, Hongshan District, Wuhan, 430070, Hubei, China

    Shaokui Yi, Li-Fang Liu, Lai-Fang Zhou, Bo-Wen Zhao, Wei-Min Wang & Ze-Xia Gao

  2. Engineering Research Center of Green development for conventional aquatic biological industry in the Yangtze River Economic Belt, Ministry of Education, Wuhan, 430070, China

    Shaokui Yi, Li-Fang Liu, Lai-Fang Zhou, Bo-Wen Zhao, Wei-Min Wang & Ze-Xia Gao

  3. Engineering Technology Research Center for Fish Breeding and Culture in Hubei Province, Wuhan, 430070, China

    Ze-Xia Gao

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  1. Shaokui Yi
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Correspondence to Ze-Xia Gao.

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Yi, S., Liu, LF., Zhou, LF. et al. Screening of Biomarkers Related to Ovarian Maturation and Spawning in Blunt Snout Bream (Megalobrama amblycephala) Based on Metabolomics and Transcriptomics. Mar Biotechnol 22, 180–193 (2020). https://doi.org/10.1007/s10126-019-09943-5

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