Type F scavenger receptor expressed by endothelial cells (SREC)-II from Epinephelus coioides is a potential pathogen recognition receptor in the immune response to Vibrio parahaemolyticus infection.

Scavenger receptors play a central role in defending against infectious diseases in mammals. However, the function of SRECII remains unknown in teleost fish. In this study, type F scavenger receptor expressed by endothelial cells-II (SRECII) cDNA sequence was first identified from Epinephelus coioides, named EcSRECII, which contained an N-terminal signal peptide, eight EGF/EGF-like cysteine-rich motifs and a C-terminal low-complexity region. The gene location maps revealed that EcSRECII has the conservation of synteny among selected species. Subcellular localization showed that EcSRECII was mainly located in the cytoplasm in HEK293T cells and GS cells. In healthy E. coioides, EcSRECII mRNA was highly expressed in spleen, skin, gill, thymus and head kidney. The relative EcSRECII mRNA expression after Vibrio parahaemolyticus infection was significantly up-regulated at 12 h in spleen, head kidney and thymus, but downregulated at 1 d in skin and reduced at 3 d and 1 w in spleen. Furthermore, overexpression of EcSRECII activated NF-κB and IFN-ß signaling pathway in vitro. Taken together, these results indicated that EcSRECII could be as the potential pathogen recognition receptor for involving in bacterial infection by regulating innate immunity responses in E. coioides.

Expression profiles of dmrts and foxls during gonadal development and sex reversal induced by 17α-methyltestosterone in the orange-spotted grouper.

The orange-spotted grouper, Epinephelus coioides, is a marine protogynous hermaphrodite fish of commercial importance. There are many examples of sex change species among marine fish, but the molecular basis for the sex change is still unknown. Gonadal expression patterns of the dmrts and foxls genes in E. coioides have pointed to sexual dimorphism in this species and it has been shown that mRNA levels of dmrts and foxls to vary significantly during reproduction cycles. The steroid 17α-methyltestosterone was used to induce sex reversal in these fish, during which dmrts and foxls levels changed significantly and subsequently reverted to normal when 17α-methyltestosterone was withdrawn. Interestingly, the expression of dmrt2b and dmrt3 was not affected by this steroid. We speculate that the role of foxl2 in reproduction may be conserved via regulation of early differentiation of the ovary by the hypothalamus-pituitary-gonad axis, and dmrt2 may have a significant role in premature ovarian differentiation and maintenance in E. coioides. dmrt1 and foxl3 played a role in the development of the testes and are believed to be potential male regulatory genes.

Comparison of species-specific qPCR and metabarcoding methods to detect small pelagic fish distribution from open ocean environmental DNA.

Environmental DNA (eDNA) is increasingly used to noninvasively monitor aquatic animals in freshwater and coastal areas. However, the use of eDNA in the open ocean (hereafter referred to OceanDNA) is still limited because of the sparse distribution of eDNA in the open ocean. Small pelagic fish have a large biomass and are widely distributed in the open ocean. We tested the performance of two OceanDNA analysis methods-species-specific qPCR (quantitative polymerase chain reaction) and MiFish metabarcoding using universal primers-to determine the distribution of small pelagic fish in the open ocean. We focused on six small pelagic fish species (Sardinops melanostictus, Engraulis japonicus, Scomber japonicus, Scomber australasicus, Trachurus japonicus, and Cololabis saira) and selected the Kuroshio Extension area as a testbed, because distribution of the selected species is known to be influenced by the strong frontal structure. The results from OceanDNA methods were compared to those of net sampling to test for consistency. Then, we compared the detection performance in each target fish between the using of qPCR and MiFish methods. A positive correlation was evident between the qPCR and MiFish detection results. In the ranking of the species detection rates and spatial distribution estimations, comparable similarity was observed between results derived from the qPCR and MiFish methods. In contrast, the detection rate using the qPCR method was always higher than that of the MiFish method. Amplification bias on non-target DNA and low sample DNA quantity seemed to partially result in a lower detection rate for the MiFish method; the reason is still unclear. Considering the ability of MiFish to detect large numbers of species and the quantitative nature of qPCR, the combined usage of the two methods to monitor quantitative distribution of small pelagic fish species with information of fish community structures was recommended.

The Administration of Cortisol Induces Female-to-Male Sex Change in the Protogynous Orange-Spotted Grouper, Epinephelus coioides.

In this study, we injected cortisol into the protogynous orange-spotted grouper (Epinephelus coioides) to investigate the role of this hormone in sex change. Following injection, we evaluated gonadal changes, serum levels of steroid hormones, and sex-related gene expression during the processes of cortisol-induced sex change and cortisol withdrawal in the orange-spotted grouper. Cortisol treatment caused the degeneration of oocytes and induced sex change in a dose-dependent manner. Over the long-term, we observed a significant increase in serum 11-ketotestosterone (11-KT) levels in all cortisol-treated groups, although levels of 17ß-estradiol did not change significantly. Consistent with the elevation of serum 11-KT levels, the expression of genes related to testicular development was also significantly up-regulated in the cortisol-treated groups. Based on our results, we propose that cortisol may trigger masculinization by inducing the synthesis of 11-KT and by directly activating the expression of sex-related genes. Furthermore, we found that cortisol-induced sex change was not permanent and could be reversed after the withdrawal of cortisol treatment.

Overexpression of Anti-müllerian Hormone Gene in vivo Affects Gonad Sex Differentiation in Undifferentiated Orange-Spotted Groupers (Epinephelus coioides).

Sex differentiation in teleost fishes occurs in response to sex determination signals, which induce the gonad to develop as either an ovary or testis. However, sex differentiation mechanisms in fishes are diverse, and information on gonad differentiation in sex changing fishes remains limited. The orange-spotted grouper (Epinephelus coioides) is a protogynous hermaphroditic fish that provides an ideal model for investigating gonad differentiation in vertebrates. In this study, Transcriptome data showed that expression levels of amh and amhrII in gonads were increased during sex differentiation. Then we investigated the effect of overexpression anti-Müllerian hormone (Amh) on gonad development in juvenile orange-spotted groupers. Expression levels of female-related genes and serum 17ß-estradiol levels were decreased, while expression of male-related genes and serum 11-ketotestosterone levels were increased in fish fed with amh-plasmid. Overexpression of Amh was also promoted the spermatogonia proliferation and induced the development of male gonads in undifferentiated orange-spotted groupers, but that this male tendency was preceded by female differentiation. In summary, these results illustrated that Amh overexpression by amh-plasmid feeding induced male gonad development in undifferentiated groupers.

New Insights Into the Role of Follicle-Stimulating Hormone in Sex Differentiation of the Protogynous Orange-Spotted Grouper, Epinephelus coioides.

Follicle-stimulating hormone (FSH) signaling is considered to be essential for early gametogenesis in teleosts, but its functional roles during sex differentiation are largely unknown. In this study, we investigated the effects of long-term and short-term FSH injection on sex differentiation in the protogynous orange-spotted grouper (Epinephelus coioides). Long-term FSH treatment initially promoted the formation of ovaries but subsequently induced a male fate. The expression of female pathway genes was initially increased but then decreased, whereas the expression of male pathway genes was up-regulated only during long-term FSH treatment. The genes related to the synthesis of sex steroid hormones, as well as serum 11-ketotestosterone and estradiol, were also up-regulated during long-term FSH treatment. Short-term FSH treatment activated genes in the female pathway (especially cyp19a1a) at low doses but caused inhibition at high doses. Genes in the male pathway were up-regulated by high concentrations of FSH over the short term. Finally, we found that low, but not high, concentrations of FSH treatment activated cyp19a1a promoter activities in human embryonic kidney (HEK) 293 cells. Overall, our data suggested that FSH may induce ovarian differentiation or a change to a male sex fate in the protogynous orange-spotted grouper, and that these processes occurred in an FSH concentration-dependent manner.

The next-generation sequencing reveals the complete mitochondrial genome of Alosa sapidissima (Perciformes: Clupeidae) with phylogenetic consideration.

The complete mitochondrial genome of the Alosa sapidissima is presented in this study. The mitochondrial genome is 16,741 bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and a control region. The gene order and composition of Epinephelus awoara mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand in descending order is 28.79% of G, 28.39% of T, 24.94% of A, and 17.88% of C. With the exception of the NADH dehydrogenase subunit 6 (ND6) and 8 tRNA genes, all other mitochondrial genes are encoded on the heavy strand. The phylogenetic analysis by maximum-likelihood (ML) method showed that the A. sapidissima has the closer relationship to between the A. alosa and A. pseudoharengus in the phylogenetic relationship.

The complete mitochondrial genome of the hybrid grouper (Cromileptes altivelis♀ × Epinephelus lanceolatus♂) with phylogenetic consideration.

The complete mitochondrial genome of the Hybrid grouper (Cromileptes altivelis♀ × Epinephelus lanceolatus♂) was presented in this study. The mitochondrial genome is 16,501 bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and a control region. The gene order and composition of Hybrid grouper mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 26.24% of A, 15.68% of C, 29.07% of T, and29.01% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand. The phylogenetic analysis by maximum-likelihood (ML) method shows that the hybrid grouper has closer relationship to C. altivelis.