Hyriopsis Cumingii Research Articles

Identification of wnt2 in the pearl mussel Hyriopsis cumingii and its role in innate immunity and gonadal development

Wnt2 is a significant factor in the Wnt signaling pathway, which is associated with a variety of physiological activities, including inflammatory response, cell apoptosis, reproductive system development, and cell differentiation. Hyriopsis cumingii is the main pearl breeding mussel in China. However, the role of wnt2 in this species remains unclear. In this study, wnt2 from H. cumingii was cloned and identified. The full-length cDNA of wnt2 is 1524 bp, containing a 963 bp open reading frame (ORF), encoding 320 amino acid residues. The tissue distribution of H. cumingii indicated that wnt2 was predominantly highly expressed in the ovary and gill. And the expression profile after Aeromonas hydrophila or LPS injection indicated that wnt2 was up-regulated in gill, suggesting its role in the innate immune response. The expression of wnt2 was high at 4-month-old of early gonadal development and throughout ovarian development. In situ hybridization (ISH) showed significant hybridization signals on the gills and mature eggs of female gonads. In addition, miR-1988b-5p was found to negatively regulate wnt2 to affect the expression of key genes (frizzled-5, ctnnb1, and tcf7l) in the Wnt signaling pathway. Thus, these findings suggest a key role for wnt2 in immune regulation and gonadal development in H. cumingii.

The temptin gene of the clade Lophotrochozoa is involved in formation of the prismatic layer during biomineralization in molluscs

The biomineralization mechanism of mollusc shell has been studied for a long time, but there is a lack of understanding about the relationship between the shell formation in vitro and the signaling system in vivo. In this study, we cloned a novel shell matrix protein gene (hc-temptin), which only be characterized as a water-borne protein pheromone of molluscs in previous studies, from the freshwater mussel Hyriopsis cumingii. By bioinformatics analysis we found that temptin was a gene unique to the clade Lophotrochozoa, and it exists in all mollusc taxa except Cephalopoda. The current data supported the premise that temptin was generated in the early emergence of molluscs and that it maintained a high mutation rate to evolve relative independently. The specificity of hc-temptin expression in the mantle tissue suggests its potential to participate in biomineralization. Its sequence contained typical Ca2+ binding sites. Our experiments involving the pearl formation process, damaged shell repair process, and RNAi experiment showed that hc-temptin was a shell matrix protein that plays an important role in formation of the prismatic layer. The results of this study provided new insights about the origin of the temptin gene and its role in molluscs.

Effect of temperature on pearl deposition rate and matrix protein gene expressions in Hyriopsis cumingii

In this study, we investigated the impact of water temperature on pearl formation and the molecular mechanisms underlying this biomineralization process in the mussel Hyriopsis cumingii. In the laboratory experiment, recipient mussels received saibo grafts from donor mussels and then were cultivated at five temperatures (16, 20, 24, 28, and 32 °C) for 1 month. We found that speed of pearl formation varied with water temperature: there was no visible pearl granules at 16 and 20 °C, tawny pearl granules were collected at 24 °C and lustrous pearl granules were collected at 28 and 32 °C. The expressions of several nacreous layer matrix protein genes (hic22, hic24, HcTyr Hc-upsalin and Hcperlucin) at 28 °C and 32 °C were peaked on day 28, and the expressions of other nacreous layer matrix protein genes (hic52 and hic74) on day 28 were significant higher than day 14. In the natural environment, pearl deposition rate also increased as water temperature rose, and the optimum range for pearl growth in H. cumingii was between 28.3 and 30.3 °C. Expression of the prismatic layer-related matrix protein genes hic31 and both prismatic and nacreous layer-related matrix protein gene silkmapin was significantly high in winter, and expressions of the nacreous layer-related matrix protein genes (hic24, hic74, HcTyr, Hc-upsalin and Hcperlucin) were always higher during higher temperature seasons compared with winter. These results indicate that high water temperature can speed up pearl formation and further regulate matrix protein secretion to accelerate pearl nacre deposition.

Identification and Functional Analysis of the Sex-Determiner Transformer-2 Homologue in the Freshwater Pearl Mussel, Hyriopsis cumingii.

Transformer-2 (Tra-2) is an upstream regulatory element of the sex regulation mechanism in insects and plays a critical role in sex formation. To understand the role of tra-2 in Hyriopsis cumingii, the full-length Hctra-2 (1867 bp) was obtained from the gonads, and sequence alignment with other species showed that HCTRA-2 protein had a highly conserved RRM domain. Phylogenetic analysis showed that the HCTRA-2 protein was a close relative to of the mollusks TRA-2 protein. The qRT-PCR of tissue-specific expression pattern showed that the Hctra-2 was abundant in gonads, and the expression in testes was higher than that in ovaries (p < 0.01). It suggests that Hctra-2 may play a potential regulatory role in gonadal development of H. cumingii. In the early gonadal development, the Hctra-2 expression was the highest on the third day after fertilization and increased slightly from 4 months to 5 months, which may be related to the embryonic sex determination and early gonadal development. In situ hybridization showed that Hctra-2 mRNA signals were present in both male and female gonads. After silencing Hctra-2 by RNAi, the expression levels of Hcfem-1b and Hcdmrt were changed. It is speculated that there may be a certain relationship between them, which plays an important role in the sex regulation of H. cumingii. Our research will help to deepen our understanding of the shellfish sex determination mechanisms.

Effects of chitosan oligosaccharide and hyriopsis cumingii polysaccharide on the quality of wheat flour and extruded flour products

Effects of chitosan oligosaccharide (COS) and hyriopsis cumingii polysaccharide (HCP) on the quality of wheat flour and corresponding extruded flour products were investigated in this work. The results showed that both COS and HCP are conducive to the improvement of dough quality. Moreover, compared to control group samples, the moisture content, expansion ratio and oil absorption rate of the samples were increased and the hardness were decreased with the addition of COS. These phenomena indicate the quality of extruded flour products became better in the presence of COS as well. However, HCP has little or no effect on the quality of extruded flour products may be due to its degradation under high temperature and pressure extrusion. COS with higher stability exhibited better improvement effects on the quality of extruded flour products and showed a promising prospect for application in extruded food industry.

Histopathology, antioxidant responses, transcriptome and gene expression analysis in triangle sail mussel Hyriopsis cumingii after bacterial infection

Bacterial disease outbreaks in filter feeder bivalve Hyriopsis cumingii as water contamination become more frequent in the water ecosystem, especially in intensive aquaculture habitats. To characterize host-pathogen interactions between H. cumingii and bacterial infection, we investigated the effects of Stenotrophomonas maltophilia HOP3 and Aeromonas veronii GL1 on the antioxidant response, tissue invasion and transcriptome expression of H. cumingii by infectivity trials. We showed that bacterial infections resulted in tubular necrosis of the hepatopancreas and induced the acute immune response in H. cumingii. The transcriptomic study identified a total of 5957 differentially expressed genes (DEGs) after A. veronii challenge. These DEGs were implicated in 302 KEGG pathways, notably in Apoptosis, Phagosome and Lysosome. The results showed that the relative expressions of all six immune-related DEGs were effectively stimulated with A. veronii, accompanied by tissue differences. Overall, these findings will contribute to an analysis of the immune response of H. cumingii to bacterial infection at the transcriptomic level and its genomic resource for research.

Cloning of a HcCreb gene and analysis of its effects on nacre color and melanin synthesis in Hyriopsis cumingii.

Creb (Cyclic AMP response element binding protein) is a nuclear regulatory factor that regulates transcription through autophosphorylation. In melanocytes, cAMP’s corresponding elements bind to the Creb protein to autophosphorylation and activate MITF (Microphthalmia-associated transcription factor). MITF stimulates Tyrosine(tyr) to induce melanocytes to differentiate into eumelanin and pheomelanin. In this study, a HcCreb gene in Hyriopsis cumingii was cloned and its effects on melanin synthesis and nacre color were studied. HcCreb was expressed in both purple and white mussels, and there was a significant difference in expression between adductor muscle (p<0.01) and mantle tissue (p<0.05). Other tissues did not show significant differences (except for gill tissue), and in general, the level of mRNA expression was higher in purple mussels than in white mussels. In both white and purple mussels expression levels in gill tissue was the highest, followed by the mantle. Strong and specific mRNA signals were detected in the dorsal epithelial cells of the mantle pallial layer, indicating that HcCreb may be involved in nacre formation. After arbutin treatment, the expression of HcCreb decreased significantly. By further testing the changes in mantle melanin content it was found that the melanin content after arbutin treatment decreased significantly compared to the control group (p<0.05). It is speculated that the HcCreb gene plays a role in the process of melanin synthesis and nacre color formation in H. cumingii.

Molecular characterization of the nitric oxide synthase gene and its immunomodulation of nitric oxide production in the triangle shell mussel (Hyriopsis cumingii)

Nitric oxide synthase (NOS) is a critical enzyme that catalyzes nitric oxide biosynthesis and orchestrates various immunological responses mediated by nitric oxide (NO) in host animals. In this study, the NOS gene was identified in the triangle shell mussel (Hyriopsis cumingii) (HcNOS). HcNOS was highly conserved in the characteristic gene structures of NOS. Phylogenetic analysis suggested that HcNOS was a typical invertebrate NOS. Further gene expression analysis, NOS activity assays and nitric oxide content measurements demonstrated the inducibility of HcNOS in responses to lipopolysaccharide (LPS) challenge and during tissue transplantation. Of note, mantle grafting induced a prolonged HcNOS/NO response, suggesting that through the HcNOS/NO system, multiple immunomodulators may play decisive roles in tissue grafting in triangle shell mussels. Thus, HcNOS appears to be a crucial player in responding to both bacterial infection and tissue transplantation.

Control of cyanobacterial blooms in different polyculture patterns of filter feeders and effects of these patterns on water quality and microbial community in aquacultural ponds

Six experimental groups were established for a land-based enclosure experiment: Macrobrachium rosenbergii Monoculture group; M. rosenbergii and Hyriopsis cumingii polyculture group; M. rosenbergii and silver and bighead carp polyculture group; and M. rosenbergii, H. cumingii, and silver and bighead carp polyculture groups (GSBH1, GSBH2, and GSBH3) in different densities of H. cumingii. Each group had four replicates. Control of cyanobacterial blooms in different polyculture patterns and the effects of these patterns on water quality and microbial community were studied. Results showed that the silver and bighead carp polyculture could control cyanobacterial blooms. The H. cumingii and silver and bighead carp polyculture improved water quality, and the GSBH2 group had the best purifying effect. Moreover, different microbial communities were observed in the different polyculture patterns. The silver and bighead carp polyculture exerted obvious effect on microbial community, whereas H. cumingii only slightly affected microbial community. Water quality parameters that significantly influenced microbial community included Secchi depth transparency, total nitrogen content, total phosphorus content, total dissolved phosphate content, nitrite nitrogen content, chemical oxygen demand, and chlorophyll a content. Thus, microbial community could also serve as an indicator of nutrient levels of water bodies.

The mantle exosome proteins of Hyriopsis cumingii participate in shell and nacre color formation

Pearl color is closely related to the nacre color of shell in Hyriopsis cumingii, and pearl color has a huge impact on its price. The nacre is an important part of the shell, and studies have suggested that mantle exosomes participated in shell formation. Exosomes contain many different proteins that are involved in different biological processes. In this study, exosomes were extracted from mantles of mussels with different nacre color. TMT quantitative proteome sequencing analysis was performed on purple and white mussel mantle exosomes, and 4861 proteins were obtained. Based on the standard of (|log2 (Fold change)| ≥ 1.2 or ≤ 0.83 and p-value ≤ 0.05), a total of 758 differentially expressed proteins were found. Some proteins involved in shell and nacre color formation were predicted with the proteins annotate, GO classification system. Moreover, 14 differentially expressed proteins (including eight up-regulated proteins and six down-regulated proteins) were validated using parallel reaction monitoring (PRM) assays. Overall, this information will be useful to improve our understanding of the molecular mechanisms of shell and nacre color formation in H. cumingii.

Bioaccumulation, Metabolism, and Biomarker Responses in Hyriopsis cumingii Exposed to 4‐Mono‐Chlorinated Dibenzothiophene

Polychlorinated dibenzothiophenes (PCDTs) are sulfur analogues of polychlorinated dibenzofurans with prevalent occurrence in aquatic environments and potential ecological risks. However, data on the behavior and toxicity of PCDTs in aquatic organisms remain scarce. In the present study, the bioaccumulation, metabolism, and oxidative damage of 4-mono-chlorinated dibenzothiophene (4-mono-CDT) in freshwater mussel (Hyriopsis cumingii) were investigated after exposure to 4-mono-CDT in semistatic water. The uptake rates, depuration rates, half-lives, and bioconcentration factors of 4-mono-CDT in hepatopancreas, gill, and muscle tissues ranged from 0.492 to 1.652 L d-1 g-1 dry weight, from 0.117 to 0.308 d-1 , from 2.250 to 5.924 d, and from 2.903 to 8.045 × 103 L kg-1 dry weight, respectively. A dechlorinated metabolite (dibenzothiophene) was detected in hepatopancreas tissue, indicating that dechlorination was the main metabolic pathway of 4-mono-CDT. As the exposure time increased, the activities of superoxide dismutase, catalase, and glutathione peroxidase were induced or inhibited in the different experimental groups. The malondialdehyde content increased with increasing 4-mono-CDT dose and exposure time. A higher concentration of 4-mono-CDT corresponded to a greater integrated biomarker response in each tissue and greater oxidative damage. The antioxidant enzymes in hepatopancreas were more sensitive to 4-mono-CDT than those in gill. The results provide useful information on the behavior and ecotoxicity of PCDTs in freshwater mussels. Environ Toxicol Chem 2021;40:1873-1882. © 2021 SETAC.

Bacterial community in gut, shell sediment, and surrounding water of Hyriopsis cumingii fed three different microalgal diets

A challenge in the intelligent breeding of pearl mussels (Hyriopsis cumingii) is the lack of a high-quality microalgal diet. To select the optimal microalgae diet, we examined the effect of feeding of different microalgal diets on the bacterial community composition in the gut, shell surface sediment, and surrounding water of pearl mussels. The bacterial communities of these three samples were analyzed using Illumina high-throughput sequencing. The effect of feeding of three types of microalgae (Scenedesmus abundans, Chlorella pyrenoidosa, and Scenedesmus dimorphus) to triangle sail mussels was investigated in a small intelligent aquaculture system. The different microalgal diets affected the growth of H. cumingii, and optimal growth was observed in the S. dimorphus group (P < 0.05). The three microalgal diets significantly affected the microbiota abundance and diversity in the gut, surface sediment, and surrounding water (P < 0.05). We also explored the bacterial community relationship among the gut, surface sediment, and surrounding water. A strong correlation was observed between bacterial community in the surface sediment and surrounding water. Our finding that different microalgal diets can affect the bacterial community of H. cumingii gut, shell surface sediment, and surrounding water expand the knowledge of the bacterial community in aquaculture ecosystems of mussels and may contribute to establishing an intelligent breeding model for H. cumingii.

Effects of partial substitution of NaCl on myofibrillar protein properties from pearl mussel Hyriopsis cumingii muscle: Structural characteristics and aggregation behaviors

The effects of NaCl and its partial substitutes (KCl, MgCl2 and CaCl2) on solubility, structural characteristics and aggregation behaviors of myofibrillar protein (MP) from pearl mussel muscle were investigated and compared. MP at 0.6 M NaCl was beneficial to protein unfolding and showed excellent potential functional properties. When NaCl was substituted in low level, MPs also showed good solubility and ordered microstructure as well as NaCl, especially MgCl2 and CaCl2, due to the unfolding of α-helical structures and subsequently exposed tyrosine residues and hydrophobic groups. However, the obviously increased disulfide bonds and hydrophobic interactions in high substitution level indicated the excessive non-sodium salts had negative effects on molecular rearrangement, leading to irregular and overly tight of microstructure. Thus, NaCl partially substituted by KCl, MgCl2 and CaCl2 in low substitution level is promising to improve functional properties of MP in low-sodium meat products.

Dorsal regulates the expression of two phage lysozymes acquired via horizontal gene transfer in triangle sail mussel Hyriopsis cumingii

Dorsal is a Rel/NF-κB transcription factor, which forms a key part of the Toll pathway. Lysozyme is a ubiquitous enzyme that degrades bacterial cell walls. In this study, a Dorsal homolog was cloned and characterized from triangle sail mussel Hyriopsis cumingii, namely, HcDorsal. Dorsal consisted of 3041 bp, including a 1938 bp open reading frame encoding a 645 amino acid protein. The deduced HcDorsal protein contained a Rel homology domain and an Ig-like, plexin, transcription factor domain. Analysis of expression patterns showed that HcDorsal was highly expressed in the hepatopancreas of H. cumingii. The expression level of HcDorsal continuously increased after Vibrio parahaemolyticus stimulation. When HcDorsal was knocked down by siRNA interference, two phage lysozyme genes (HcLyso1 and HcLyso2) obtained by horizontal gene transfer were significantly downregulated in hemocytes of mussels. Furthermore, knockdown of HcLyso1 and HcLyso2 could weaken V. parahaemolyticus clearance ability. Recombinant HcLyso1 and HcLyso2 proteins accelerated the bacterial clearance in vivo in mussels and evidently inhibited the growth of V. parahaemolyticus. These results suggested that HcDorsal could be activated after V. parahaemolyticus stimulation and then modulate the immune response through the transcriptional regulation of HcLyso1 and HcLyso2, thereby playing a protective role in mussels.

Effects of Acute Ammonia Stress on Antioxidant Responses, Histopathology and Ammonia Detoxification Metabolism in Triangle Sail Mussels (Hyriopsis cumingii)

Ammonia is one of the major pollutants in the aquatic ecosystem. Hyriopsis cumingii has great potential for the restoration of eutrophic water. However, there is no study investigating the effect of ammonia exposure in H. cumingii. The median lethal concentration (96 h LC50) of unionized ammonium was 12.86 mg/L in H. cumingii. In the study, H. cumingii were exposed to 6.43 mg L−1 unionized ammonium (1/2 96 h LC50) for 0, 6, 12, 24, 48, 72, and 96 h. High environment ammonia induced antioxidant response to protect the body from oxidative damage. After exposure to ammonia, there was a same trend of induction followed by inhibition of the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione-S-transferases (GST) in the hepatopancreas and gills of H. cumingii. However, the antioxidant response could not completely counteract the oxidation effect during the exposure period, resulting in lipid peroxidation (LPO) and tissue injury in the hepatopancreas and gills of H. cumingii eventually. Moreover, this study indicated that glutamine synthetase (GS), glutamate dehydrogenase (GDH), alanine aminotransaminase (ALT), and aspartate aminotransaminase (AST) in the hepatopancreas and gills may play an important role in ammonia detoxification of H. cumingii. Our results will be helpful to understand the mechanism of aquatic toxicology induced by ammonia in shellfish.

Ecological Efficiency of the Mussel Hyriopsis cumingii (Lea, 1852) on Particulate Organic Matter Filtering, Algal Controlling and Water Quality Regulation

Hyriopsis cumingii plays an important role in aquatic ecosystems and fishery economy due to its potential value for water purification and edibleness. The present study was undertaken to reveal the short- and long-term purifying effects of H. cumingii on pond water. The short-term experiment results showed that total suspended solids, particulate organic carbon, and fatty acid in water were significantly eliminated by H. cumingii, with average percentage of filtering effect respectively reaching 22.19%, 57.48%, and 21.00% in the high-density group. Analogously, H. cumingii could significantly reduce phytoplankton biomass, species number, density, and chlorophyll-a concentration, especially in the control of diatoms, green algae, and cyanobacteria. Besides, electric conductivity also was significantly reduced by H. cumingii and its variation tendency showed a typical density-dependent effect. Similar purification effects were observed in chemical oxygen demand (COD), total nitrogen, and total phosphorus. In the long-term monitoring test, H. cumingii has also shown positive effects on the water transparency and the control to phytoplankton and COD in recirculating aquaculture pond. Overall, H. cumingii showed excellent ecological function, and the mixed breeding density of shellfish in the recirculating aquaculture pond is worth further exploration.

Integrated mRNA and miRNA expression profile analysis of female and male gonads in Hyriopsis cumingii

Hyriopsis cumingii is an important species for freshwater pearl cultivation in China. In terms of pearl production, males have larger pearls and better glossiness than females, but there are few reports focusing on the sex of H. cumingii. In this study, six mRNA and six microRNA (miRNA) libraries were prepared from ovaries and testes. Additionally, 28,502 differentially expressed genes (DEGs) and 32 differentially expressed miRNAs (DEMs) were identified. Compared with testis, 14,360 mRNAs and 20 miRNAs were up-regulated in ovary, 14,142 mRNAs and 12 miRNAs were down-regulated. In DEGs, the known genes related to sex determinism and/or differentiation were also identified, such as DMRT1, SOX9, SF1 for males, FOXL2 for females, and other potentially significant candidate genes. Three sex-related pathways have also been identified, which are Wnt, Notch, and TGF-beta. In 32 DEMs, the three miRNAs (miR-9-5p, miR-92, miR-184) were paid more attention, they predicted 28 target genes, which may also be candidates for sex-related miRNAs and genes. Differential miRNAs target genes analysis reveals the pathway associated with oocyte meiosis and spermatogenesis. Overall, the findings of the study provide significant insights to enhance our understanding of sex differentiation and/or sex determination mechanisms for H. cumingii.

Characteristics of a Critical Calcium Transportation Regulator: Plasma Membrane Ca&amp;lt;sup&amp;gt;2+&amp;lt;/sup&amp;gt;-ATPase Involved in Calcium Homeostasis from &amp;lt;i&amp;gt;Hyriopsis cumingii&amp;lt;/i&amp;gt; (Lea)

Plasma Membrane Calcium ATPase (PMCA) plays a critical role in transporting Ca2+ out of the cytosol across the plasma membrane. Here, a full-length cDNA sequence of plasma membrane Ca2+-ATPase gene was isolated from the gill of Hyriopsis cumingii (HcPMCA) by using SMART RACE technique. The entire cDNA was 5230 bp, including a 417-bp 5'-UTR, a 3588-bp ORF and a 1225-bp 3'-UTR, encoding a 1195-amino acid protein, and no putative signal peptide was predicted. Compared with PMCA homologs from seawater mollusks, HcPMCA had high similarity with them in both sequence and structure. Tissue-specific expression analysis revealed that HcPMCA mRNA was detected in all the sampled tissues, but was prominently expressed in the gill and mantle. When exposed to a serie of increasing Ca2+ that lasted for 7 days, the mRNA expression of HcPMCA in the mantle was slightly downregulated, but peaked at 60 mg/L. Moreover, the temporal expression of HcPMCA transcripts in the mantle after 60 mg/L Ca2+ exposure was shown to be bell-shaped, which was slightly downregulated at 24 h, but upregulated from 24 h to 48 h post-treatment, peaking at 48 h. The result of present study provides useful information for further studies on function and regulation mechanism of HcPMCA gene.

Expression and functional characterization of the CUB domain-containing protein from the triangle sail mussel (Hyriopsis cumingii) in response to pathogenic infection.

The complement C1r/C1s, Uegf, and Bmp1 (CUB) domains, which are most exclusively found in extracellular and plasma membrane-related proteins, are involved in various biological processes. In this study, a CUB domain-containing protein (designed as HcCDCP) was cloned and characterized from freshwater pearl mussel (Hyriopsis cumingii). The 2280bp complete cDNA of the HcCDCP contained a 1002bp open reading frame, which encoded a protein with 333 amino acids. The predicted HcCDCP protein contained a typical CUB domain and a transmembrane region. The tissue distribution analysis indicated that the HcCDCP was detected in all tissues, and the highest expression was found in hepatopancreas followed by gills. After infection with bacteria (i.e., Staphylococcus aureus and Vibrio parahaemolyticus), virus (white spot syndrome virus) and virus analogs (poly[I:C]), the mRNA level of the HcCDCP was significantly upregulated, suggesting that the HcCDCP might be involved in host immune defense response. The RNA interference revealed that the silencing of the HcCDCP could evidently inhibit the expression levels of lysozyme and tumor necrosis factor. Moreover, the recombinant protein of the CUB domain (rCUB) possessed binding capacity to eight different kinds of bacteria. The polysaccharide binding assay showed that the rCUB specifically bound to lipopolysaccharide, peptidoglycan, and D-mannose. This study provided valuable information for exploring the biological roles of CDCPs in the host defense system of mollusks.

Comparative proteomic study on fem-1b in female and male gonads in Hyriopsis cumingii

Sex regulation of bivalves is a complex process, and there has also been little research on their sex-determining genes. To better understand the sex determination/differentiation of bivalves, we analyzed the gonadal proteomics of Hyriopsis cumingii. We identified 673 peptides and 76 proteins. The 35 differentially expressed proteins identified included 19 upregulated and 16 downregulated proteins. FEM-1B protein associated with sex determination showed female-specific expression; the function of fem-1b gene was explored. The full-length cDNA of fem-1b cloned by RACE was 2105 bp. Quantitative PCR (qPCR) was performed on male and female tissues and early developmental stages; the results showed that fem-1b was expressed in both male and female tissues, and its expression level in gonads was significantly higher in females than in males (P < 0.01). In early embryonic development, the fem-1b mRNA expression increased significantly on the third day after fertilization (cleavage stage). Results of in situ hybridization showed a strong signal on oocyte membranes and no signal in male gonads. Our results indicate that fem-1b plays a certain role in sex determination and oocyte development of H. cumingii. This study provides valuable resources for understanding the mechanism and pathway of sex determination/differentiation of H. cumingii.