Project description:microRNAs (miRNAs) are an evolutionarily conserved class of non-coding RNA molecules, which regulate kinds of biological processes at post-transcriptional level. Investigation of miRNAs expression profiles using high-throughput strategies is efficiently conductive to identify and characterize miRNAs. In this study, through Solexa deep sequencing approach, we obtained 115 orange spotted grouper (Epinephelus coioides) encoded miRNAs. Among them, 107 miRNAs shared high similarity with miRNAs encoded by zebrafish (Danio rerio) and other four vertebrates, indicating that cellular miRNAs are highly conserved between species.
Project description:microRNAs (miRNAs) are an evolutionarily conserved class of non-coding RNA molecules, which regulate kinds of biological processes at post-transcriptional level. Investigation of miRNAs expression profiles using high-throughput strategies is efficiently conductive to identify and characterize miRNAs. In this study, through Solexa deep sequencing approach, we obtained 115 orange spotted grouper (Epinephelus coioides) encoded miRNAs. Among them, 107 miRNAs shared high similarity with miRNAs encoded by zebrafish (Danio rerio) and other four vertebrates, indicating that cellular miRNAs are highly conserved between species. 18-26 nt small RNAs from GS cells were sequenced in one Solexa lane
Project description:Sex differentiation is a complex process that requires many factors to regulate proliferation, differentiation, development, and organization of the gonads. In teleosts, the molecular mechanisms of sex differentiation are diverse and unclear, especially in hermaphrodite fish. In the present study, 15 cell types, including germ cells, follicle cells, and theca cells, and so on, were identified using single-cell RNA sequencing (scRNA-seq) in the gonads during sex differentiation in a hermaphrodite fish, orange-spotted grouper (Epinephelus coioides). Two pre-follicle cell types were defined, and the differentiating trajectories of the follicle cells were outlined. Notably, both pre-follicle and follicle cells highly expressed male-related genes synchronously, such as amh, sox9, and dmrt3 in pre-follicle cells, as well as dmrt1 in follicle cells. Oocyte possessed two distinct states, with high expression of oocyte development related genes in one state and spermatogenesis related genes in the other state, respectively. Our results provide novel insights into cell types and lineage tracing in the gonads during sex differentiation in hermaphrodite species.
Project description:Cortisol was injected into the protogynous epinephelus coioides to investigate the role of this hormone on sex change. Following injection, we evaluated sex-related gene expression during the processes of cortisol-induced sex change in epinephelus coioides.
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized analysis of the immune response to V. alginolyticus in Epinephelus coioides larvae, we used high-throughput deep sequencing technology to study the effect of infection on gene expression. Methods: Epinephelus coioides larvae (30-days-old) were injected with V. alginolyticus , and fish were sacrificed at 24 h after infection. Ten wild-type Epinephelus coioides larvae were used to build an EST library for transcriptome analysis. The library products were prepared for sequencing analysis using an Illumina HiSeq™ 2000. Raw data were saved as fastq files. The following were removed: reads with adaptors, reads in which over 10% bases were unknown, and low quality reads (i.e., the percentage of bases of quality value ≤ 5 exceeds 50% in the read). Clean reads were mapped to reference sequences using SOAP aligner/soap2 . The randomness of RNA fragmentation was used to construct the library, and the numbers of reads mapped to the reference sequence were calculated. The RPKM method (Reads Per kb per Million reads) was used to calculate gene expression level, and differentially expressed genes (DEGs) Results: A total of 114,851,002 reads were assembled, consisting of 9,687,355,560 nucleotides; these were further assembled into 209,082 contigs with a mean length of 372 bp. Gene ontology (GO) analysis of the transcriptome revealed 12 cellular component subcategories, 16 molecular function subcategories, and 42 biological process subcategories (P value < 0.05). A total of 32664 Epinephelus coioides genes were mapped to the Kyoto Encyclopedia of Genes and Genomes (KEGG); 1504 differentially expressed genes (DEGs) were subsequently identified, in 12 categories (P value < 0.05). Vibrio infection affected the expression of genes involved in complementation, coagulation cascades, pathogen (Staphylococcus aureus) infection, phagosome activity, antigen processing, and the antigen presentation pathway. Conclusions: We conclude that the complement pathway of innate immunity and the hepicidin antimicrobial peptide may play important roles in the defense of Epinephelus coioides larvae against V. alginolyticus, and the immune response may activate at 4h after bacterial infection. These results implicate the complement pathway signal pathway in immunity during V. alginolyticus infection at early developmental stages, enhancing our understanding of the mechanisms underlying the immune response to Vibrio infection in Epinephelus coioides.