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Front Genet
2020 Mar 04;11:161. doi: 10.3389/fgene.2020.00161.
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Beta-Hydroxysteroid Dehydrogenase Genes in Orange-Spotted Grouper (Epinephelus coioides): Genome-Wide Identification and Expression Analysis During Sex Reversal.
Xiao L, Guo Y, Wang D, Zhao M, Hou X, Li S, Lin H, Zhang Y.
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Beta-hydroxysteroid dehydrogenases (β-HSDs) are a group of steroidogenic enzymes that are involved in steroid biosynthesis and metabolism, and play a crucial role in mammalian physiology and development, including sex determination and differentiation. In the present study, a genome-wide analysis identified the numbers of β-hsd genes in orange-spotted grouper (Epinephelus coioides) (19), human (Homo sapiens) (22), mouse (Mus musculus) (24), chicken (Gallus gallus) (16), xenopus (Xenopus tropicalis) (24), coelacanth (Latimeria chalumnae) (17), spotted gar (Lepisosteus oculatus) (14), zebrafish (Danio rerio) (19), fugu (Takifugu rubripes) (19), tilapia (Oreochromis niloticus) (19), medaka (Oryzias latipes) (19), stickleback (Gasterosteus aculeatus) (17) and common carp (Cyprinus carpio) (27) samples. A comparative analysis revealed that the number of β-hsd genes in teleost fish was no greater than in tetrapods due to gene loss followed by a teleost-specific whole-genome duplication event. Based on transcriptome data from grouper brain and gonad samples during sex reversal, six β-hsd genes had relatively high expression levels in the brain, indicating that these genes may be required for neurogenesis or the maintenance of specific biological processes in the brain. In the gonad, two and eight β-hsd genes were up- and downregulated, respectively, indicating their important roles in sex reversal. Our results demonstrated that β-hsd genes may be involved in the sex reversal of grouper by regulating the synthesis and metabolism of sex steroid hormones.
FIGURE 1. Chromosome localization of 19 β-hsd genes from orange-spotted grouper.
FIGURE 2. Phylogenetic analysis, gene structure and conserved domains of β-hsd genes in orange-spotted grouper. (A) The phylogenetic tree of all β-Hsd in grouper was constructed using Neighbor-Joining method. (B) The exon/intron organization of β-hsd genes of grouper. The red boxes represent exons and black lines indicate introns. (C) The conserved protein domains in the β-Hsd were identified using CD-Search program. Each domain is indicated with a specific color. Yellow: NADB_Rossmann superfamily, green: PLN02864 superfamily, purple: SCP2 superfamily, blue: SDR superfamily.
FIGURE 3. Phylogenetic relationships of β-Hsd proteins from grouper and other five species. The phylogenetic tree was constructed using MEGA7 by Maximum Likelihood method with the amino acid sequences of β-Hsd from grouper (Epinephelus coioides, Ec), (Homo sapiens, Hs), mouse (Mus musculus, Mm), zebrafish (Danio rerio, Dr), tilapia (Oreochromis niloticus, On), and stickleback (Gasterosteus aculeatus, Ga).
FIGURE 4. Clustering of the expression profiles of 19 grouper β-hsd genes during sex reversal. Genes were clustered according to phylogenetic relationships in expression profiles. The RPKM values were transformed into Z scores. Z scores were plotted according to Z = (x - μ)/σ, where x is the log 2 transformed gene expression measurement and μ and σ are the mean and standard deviations of expression of the gene. Red and blue colors indicate high and low relative expression levels after homogenization, respectively. The RPKM value for each gene and each stage was provided in the figure.
FIGURE 5. Quantitative RT-PCR examination of 10 β-hsd genes expressions in the brain and gonad during sex reversal of grouper. Values represent the relative mRNA expression in relation to internal control (β-actin gene). Data were expressed as the mean ± SEM for four replicates. In each panel, different lowercase letters indicate significant differences (p < 0.05).
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