Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Front Endocrinol (Lausanne)
2021 Nov 11;12:756530. doi: 10.3389/fendo.2021.756530.
Show Gene links
Show Anatomy links
Application of SNP in Genetic Sex Identification and Effect of Estradiol on Gene Expression of Sex-Related Genes in Strongylocentrotus intermedius.
Han YL
,
Sun ZH
,
Chang S
,
Wen B
,
Song J
,
Zuo RT
,
Chang YQ
.
???displayArticle.abstract???
Sea urchin (Strongylocentrotus intermedius) is an economically important mariculture species in Asia, and its gonads are the only edible part. The efficiency of genetic breeding in sea urchins is hampered due to the inability to distinguish gender by appearance. In this study, we first identified a sex-associated single nucleotide polymorphism (SNP) by combining type IIB endonuclease restriction site-associated DNA sequencing (2b-RAD-seq) and genome survey. Importantly, this SNP is located within spata4, a gene specifically expressed in male. Knocking down of spata4 by RNA interference (RNAi) in male individuals led to the downregulation of other conserved testis differentiation-related genes and germ cell marker genes. We also revealed that sex ratio in this validated culture population of S. intermedius is not 1:1. Moreover, after a 58-day feeding experiment with estradiol, the expression levels of several conserved genes that are related to testis differentiation, ovary differentiation, and estrogen metabolism were dynamically changed. Taken together, our results will contribute toward improving breeding efficiency, developing sex-controlled breeding, and providing a solid base for understanding sex determination mechanisms in sea urchins.
Figure 1. The sex-associated SNP of the Strongylocentrotus intermedius. The female genotype is G/A heterozygous and the male genotype is G/G homozygous. NP, nutritive phagocytes; SPC, spermatocyte; Oo, oocyte; SPD, spermatid.
Figure 2. Sequence analysis of spata4 in S. intermedius. (A) Coding sequence and deduced amino acid sequences of spata4 in S. intermedius.
(B) Phylogenetic tree of Spata4 proteins.
Figure 3. Expression pattern of spata4 mRNA in S. intermedius. 18S was used as the control. Each bar represents mean ± standard deviation (SD) (n = 3). One-way ANOVA was used to determine statistical analysis. Asterisks (*) indicate significant differences (p ≤ 0.05).
Figure 4. RT-qPCR measured the mRNA levels of spata4 and other sex-related genes after RNAi. (A) The expression of testis differentiation genes, including spata4, dmrt1, and soxE. (B) The expression of ovary differentiation genes, including foxl2 and hsd17b8. (C) The expression of germ cell marker genes, including boule and nanos1. 18S was used as the control. The gene expression in the control group was set to 1. Each bar represents mean ± standard deviation (SD) from three individuals. Independent-sample t-test was used to perform statistical analysis. Asterisks (*) indicate significant differences (p ≤ 0.05) between knockdown and control.
Figure 5. The concentration of estradiol levels in S. intermedius. (A) Concentration of estradiol in excrement after 3 days of dietary supplementation of E2. (B) Concentration of estradiol in testis on different days after dietary supplementation of E2 (n = 6). E2, estradiol; d, day. One-way ANOVA was used to determine statistical analysis. Different letters indicate significant differences (p ≤ 0.05).
Figure 6. The significant Gene Ontology (GO) annotation of differentially expressed genes. (A) The GO cluster of upregulated genes. (B) The GO cluster of downregulated genes.
Figure 7. RT-qPCR measured the mRNA levels of sex-related genes and estrogen metabolism-related genes in the testis of E2-treated S. intermedius. (A) The expression of testis differentiation genes. (B) The expression of ovary differentiation genes. (C) The expression of germ cell marker gene. (D) The expression of steroidgenic enzyme gene. (E) The expression of estrogen metabolism gene (Emc4). 18S was used as the control. Each bar represents mean ± standard deviation (SD) (n = 3). One-way ANOVA was used to determine statistical analysis. Asterisks (*) indicate significant differences (p ≤ 0.05).
Andrew,
Exposure to 17α-ethynylestradiol causes dose and temporally dependent changes in intersex, females and vitellogenin production in the Sydney rock oyster.
2010, Pubmed
Andrew,
Exposure to 17α-ethynylestradiol causes dose and temporally dependent changes in intersex, females and vitellogenin production in the Sydney rock oyster.
2010,
Pubmed
Arizza,
Gender differences in the immune system activities of sea urchin Paracentrotus lividus.
2013,
Pubmed
,
Echinobase
Bannister,
No substantial changes in estrogen receptor and estrogen-related receptor orthologue gene transcription in Marisa cornuarietis exposed to estrogenic chemicals.
2013,
Pubmed
Barrionuevo,
SOX E genes: SOX9 and SOX8 in mammalian testis development.
2010,
Pubmed
Catchen,
Stacks: building and genotyping Loci de novo from short-read sequences.
2011,
Pubmed
Chamary,
Evidence for selection on synonymous mutations affecting stability of mRNA secondary structure in mammals.
2005,
Pubmed
Chen,
Isolation of female-specific AFLP markers and molecular identification of genetic sex in half-smooth tongue sole (Cynoglossus semilaevis).
2007,
Pubmed
Cui,
Identification of Sex-Specific Markers Through 2b-RAD Sequencing in the Sea Urchin (Mesocentrotus nudus).
2021,
Pubmed
,
Echinobase
Dan,
Genetic differentiation and efficient sex-specific marker development of a pair of Y- and X-linked markers in yellow catfish.
2013,
Pubmed
Fodor,
A critical evaluation of some of the recent so-called 'evidence' for the involvement of vertebrate-type sex steroids in the reproduction of mollusks.
2020,
Pubmed
Jiang,
Spermatogenesis associated 4 promotes Sertoli cell proliferation modulated negatively by regulatory factor X1.
2013,
Pubmed
Kamiya,
A trans-species missense SNP in Amhr2 is associated with sex determination in the tiger pufferfish, Takifugu rubripes (fugu).
2012,
Pubmed
Kikuchi,
The sex-determining locus in the tiger pufferfish, Takifugu rubripes.
2007,
Pubmed
Lai,
Xenopus Nanos1 is required to prevent endoderm gene expression and apoptosis in primordial germ cells.
2012,
Pubmed
Li,
Diverse and variable sex determination mechanisms in vertebrates.
2018,
Pubmed
Li,
SOAP2: an improved ultrafast tool for short read alignment.
2009,
Pubmed
Liu,
Sex-specific markers developed by next-generation sequencing confirmed an XX/XY sex determination system in bighead carp (Hypophthalmichthys nobilis) and silver carp (Hypophthalmichthys molitrix).
2018,
Pubmed
Liu,
Genetic manipulation of sex ratio for the large-scale breeding of YY super-male and XY all-male yellow catfish (Pelteobagrus fulvidraco (Richardson)).
2013,
Pubmed
Liu,
Cloning and characterization of testis-specific spermatogenesis associated gene homologous to human SPATA4 in rat.
2004,
Pubmed
Liu,
Molecular cloning and bioinformatic analysis of SPATA4 gene.
2005,
Pubmed
Liu,
Cloning and characterization of zebra fish SPATA4 gene and analysis of its gonad specific expression.
2005,
Pubmed
Liu,
Cloning and expression analysis of gonadogenesis-associated gene SPATA4 from rainbow trout (Oncorhynchus mykiss).
2005,
Pubmed
Liu,
[Molecular cloning of SRG2, a mouse testis spermatocyte apoptosis-related gene].
2002,
Pubmed
Matson,
DMRT1 prevents female reprogramming in the postnatal mammalian testis.
2011,
Pubmed
Mei,
Genetic basis and biotechnological manipulation of sexual dimorphism and sex determination in fish.
2015,
Pubmed
Mi,
Identification and profiling of sex-biased microRNAs from sea urchin Strongylocentrotus nudus gonad by Solexa deep sequencing.
2014,
Pubmed
,
Echinobase
Nunes,
Non-Target Effects of Green Fluorescent Protein (GFP)-Derived Double-Stranded RNA (dsRNA-GFP) Used in Honey Bee RNA Interference (RNAi) Assays.
2013,
Pubmed
Sauna,
Understanding the contribution of synonymous mutations to human disease.
2011,
Pubmed
Schwarz,
Mussels (Mytilus spp.) display an ability for rapid and high capacity uptake of the vertebrate steroid, estradiol-17β from water.
2017,
Pubmed
Scott,
Do mollusks use vertebrate sex steroids as reproductive hormones? Part I: Critical appraisal of the evidence for the presence, biosynthesis and uptake of steroids.
2012,
Pubmed
Scott,
Is there any value in measuring vertebrate steroids in invertebrates?
2018,
Pubmed
Scott,
Measurement of fish steroids in water--a review.
2007,
Pubmed
Shi,
Female-specific SNP markers provide insights into a WZ/ZZ sex determination system for mud crabs Scylla paramamosain, S. tranquebarica and S. serrata with a rapid method for genetic sex identification.
2018,
Pubmed
Sujit,
Increased DNA methylation in the spermatogenesis-associated (SPATA) genes correlates with infertility.
2020,
Pubmed
Sun,
Identification and functional characterization of piwi1 gene in sea cucumber, Apostichopus japonicas.
2021,
Pubmed
,
Echinobase
Sun,
Gonadal transcriptomic analysis and identification of candidate sex-related genes in Mesocentrotus nudus.
2019,
Pubmed
,
Echinobase
Varaksina,
[Effect of estradiol dipropionate on protein synthesis in oocytes and ovary of the sea urchin Strongylocentrotus intermedius at different stages of the reproductive cycle].
2002,
Pubmed
,
Echinobase
Wang,
Transcriptome analysis to characterize the genes related to gonad growth and fatty acid metabolism in the sea urchin Strongylocentrotus intermedius.
2019,
Pubmed
,
Echinobase
Wang,
2b-RAD: a simple and flexible method for genome-wide genotyping.
2012,
Pubmed
Warrier,
Occurrence of vertebrate steroids, estradiol 17beta and progesterone in the reproducing females of the mud crab Scylla serrata.
2001,
Pubmed
Xie,
Cloning and characterization of chicken SPATA4 gene and analysis of its specific expression.
2007,
Pubmed
Zhang,
Molecular Cloning and Sexually Dimorphic Expression Analysis of nanos2 in the Sea Urchin, Mesocentrotus nudus.
2019,
Pubmed
,
Echinobase
Zhang,
PEAR: a fast and accurate Illumina Paired-End reAd mergeR.
2014,
Pubmed
Zheng,
Evidence for the presence of sex steroid hormones in Zhikong scallop, Chlamys farreri.
2014,
Pubmed
