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A Novel Tyrosinase Gene Plays a Potential Role in Modification the Shell Organic Matrix of the Triangle Mussel Hyriopsis cumingii.

ABSTRACT: Although tyrosinases have been speculated to participate in the shell formation of mollusks, there is still a lack of experimental evidence to support this assumption. In this study, a novel tyrosinase designated HcTyr2 was isolated and characterized from the freshwater mussel Hyriopsis cumingii. The change in HcTyr2 mRNA expression during the process of embryonic development was detected by real-time quantitative PCR. The result showed that the expression of HcTyr2 mRNA was significantly upregulated at the stages of gastrulae and unmatured glochidia (P < 0.05), suggesting that this gene might fundamentally participate in the biogenesis and growth of the initial shell. Meanwhile, the upregulation of HcTyr2 mRNA at the stages of shell regeneration 24 h and 9 days after shell notching in the mantle edge (P < 0.05) implied that it might play an important role in shell periostracum and nacre formation by mediating the cross-linking of quinoproteins to promote the maturity of organic matrix. Additionally, the knockdown of HcTyr2 mRNA by RNA interference resulted in not only the suppression of periostracum growth but also structural disorder of nacre aragonite tablets, as detected by scanning electron microscopy. These results suggested that HcTyr2 might regulate the growth of shell by its oxidative ability to transform soluble matrix proteins into insoluble matrix proteins, then promoting the maturity of the shell organic framework in H. cumingii. In general, our results suggested the importance of HcTyr2 in the shell formation and regeneration of H. cumingii.

SUBMITTER: Ren G

PROVIDER: S-EPMC7045039 | biostudies-literature |

REPOSITORIES: biostudies-literature

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A C1qDC Protein (HcC1qDC6) with Three Tandem C1q Domains Is Involved in Immune Response of Triangle-Shell Pearl Mussel (Hyriopsis cumingii).

Project description:C1q-domain-containing (C1qDC) proteins are a family of proteins with a globular C1q (gC1q) domain and participate in several immune responses. In this study, a C1qDC gene was identified from the triangle-shell pearl mussel Hyriopsis cumingii (designated as HcC1qDC6). This gene has a full-length cDNA of 1782 bp and an open reading frame of 1,335 bp that encodes a 444-amino acid polypeptide containing three gC1q domains. HcC1qDC6 contains at least five exons and four introns. The mRNA transcripts of HcC1qDC6 were found to have the highest expression levels in the mantle tissue. The expression levels in the mantle and hepatopancreas were significantly upregulated by Staphylococcus aureus and Vibrio parahaemolyticus challenges. Moreover, knockdown of HcC1qDC6 inhibits the expression of two immune-related genes (tumor necrosis factor and whey acidic protein). The recombinant proteins of C1q1, C1q2, and C1q3 all exhibit a binding activity against seven bacterial species and directly bind to peptidoglycan and lipopolysaccharide. The results indicate that HcC1qDC6 is involved in the innate immunity of H. cumingii.

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Cloning and Analysis of Gene Expression of Two Toll Receptors in Freshwater Pearl Mussel Hyriopsis cumingii.

Project description:Toll receptors are involved in innate immunity of invertebrates. In this study, we identify and characterize two Toll genes (named HcToll4 and HcToll5) from triangle sail mussel Hyriopsis cumingii. HcToll4 has complete cDNA sequence of 3,162 bp and encodes a protein of 909 amino acids. HcToll5 cDNA is 4,501 bp in length and encodes a protein of 924 amino acids. Both deduced HcToll4 and HcToll5 protein contain signal peptide, extracellular leucine rich repeats (LRRs), and intracellular Toll/interleukin-1 (IL-1) receptor domains. Quantitative real-time PCR analysis revealed that HcToll4 and HcToll5 were largely distributed in the hepatopancreas and could be detected in the gills and mantle. HcToll4 and HcToll5 expression could respond to Staphylococcus aureus, Vibrio parahaemolyticus, White spot syndrome virus (WSSV) or Poly I:C challenge. RNA interference by siRNA results showed that HcToll4 and HcToll5 were involved in the regulation of theromacin (HcThe) and whey acidic protein (HcWAP) expression. Based on these results, HcToll4 and HcToll5 might play pivotal function in H. cumingii innate immune response.

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Hyriopsis cumingii

Project description:a freshwater mussel Raw sequence reads

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2022-03-17 | PXD013675 | Pride

Comparative analysis of the transcriptome in tissues secreting purple and white nacre in the pearl mussel Hyriopsis cumingii.

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Molecular cloning and characterization of Pif gene from pearl mussel, Hyriopsis cumingii, and the gene expression analysis during pearl formation.

Project description:In the present study, the Pif gene of the freshwater pearl aquaculture mussel, Hyriopsis cumingii (HcPif) was successfully cloned and functionally characterized. The full sequence of HcPif gene consists of 3415 base pairs, which putatively encode two proteins, HcPif90 and HcPif80. A sequence analysis revealed that HcPif contained a von Willebrand factor type A domain and a chitin-binding domain, and shared many functional residues with other Pif homologues. A highly conserved sequence, FKGLDEIELML, at the C-terminus of Pif80s was identified as the key functional site. The corresponding peptide fragment markedly modified the morphology of calcite crystallites in CaCO3 crystallization assay and might play an essential role in the interactive binding between HcPif80 and CaCO3. Moreover, real-time PCR results showed that HcPif gene was dominantly expressed in the pearl secreting tissues and its expression changed in response to the different development status of the pearl sac during pearl aquaculture. The gene expression of HcPif was maximum 7 days after mantle grafting and declined to about the control level on day 30. Our in vitro and in vivo experimental data indicated that HcPif gene possessed the inherent characteristics of a nacre formation gene and its expression might faithfully reflect the pearl secretion status of the pearl mussels examined. Our findings may extend the understanding of the biomineralization mechanism of nacre formation and provide a potential biomarker for pearl farming.

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An integrated transcriptome analysis of immunological responses in pearl sac of pearl mussel Hyriopsis cumingii after mantle implantation

Project description:Our findings help us gain new insight into the immunological reactions induced by transplantation in pearl mussels and markedly bring us more clues about pearl sac formation, contributing to the novel development of management strategies for pearl culture.

2019-11-30 | GSE123069 | GEO

A Novel Matrix Protein Hic31 from the Prismatic Layer of Hyriopsis Cumingii Displays a Collagen-Like Structure.

Project description:In this study, we clone and characterize a novel matrix protein, hic31, from the mantle of Hyriopsis cumingii. The amino acid composition of hic31 consists of a high proportion of Glycine residues (26.67%). Tissue expression detection by RT-PCR indicates that hic31 is expressed specifically at the mantle edge. In situ hybridization results reveals strong signals from the dorsal epithelial cells of the outer fold at the mantle edge, and weak signals from inner epithelial cells of the same fold, indicating that hic31 is a prismatic-layer matrix protein. Although BLASTP results identify no shared homology with other shell-matrix proteins or any other known proteins, the hic31 tertiary structure is similar to that of collagen I, alpha 1 and alpha 2. It has been well proved that collagen forms the basic organic frameworks in way of collagen fibrils and minerals present within or outside of these fibrils. Therefore, hic31 might be a framework-matrix protein involved in the prismatic-layer biomineralization. Besides, the gene expression of hic31 increase in the early stages of pearl sac development, indicating that hic31 may play important roles in biomineralization of the pearl prismatic layer.

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Identification and Characterization of C-Mos in Pearl Mussel Hyriopsis cumingii and Its Role in Gonadal Development.

Project description:C-Mos, a proto-oncogene, regulates oocyte maturation by activating the classical MAPK pathway in cells. To examine the function of C-Mos in Hyriopsis cumingii, C-Mos was identified in this study. The full-length cDNA of C-Mos was 2213 bp, including 144 bp in the 5' UTR, 923 bp in 3' the UTR, and 1146 bp in the open reading frame (ORF) region. During early gonad development, the expression of C-Mos from 4 to 6 months of age in H. cumingii was significantly higher than that in other months, with the highest expression in 6-month-old H. cumingii, suggesting that C-Mos may be involved in early gonadal development in H. cumingii. Clear hybridization signals were found by in situ hybridization in the oocytes, oocyte nucleus and oogonium, and a small number of hybridization signals were found in the follicular wall of the male gonads. In addition, the C-Mos RNA interference (RNAi) assay results showed that the knockdown of C-Mos caused a down-regulation of ERK and P90rsk. In summary, these results indicate that C-Mos has a crucial part to play in gonadal development in H. cumingii.

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Molecular cloning and copy number variation of a ferritin subunit (Fth1) and its association with growth in freshwater pearl mussel Hyriopsis cumingii.

Project description:Iron is one of the most important minor elements in the shells of bivalves. This study was designed to investigate the involvement of ferritin, the principal protein for iron storage, in shell growth. A novel ferritin subunit (Fth1) cDNA from the freshwater pearl mussel (Hyriopsis cumingii) was isolated and characterized. The complete cDNA contained 822 bp, with an open reading frame (ORF) of 525 bp, a 153 bp 5' untranslated region (UTR) and a 144 bp 3' UTR. The complete genomic DNA was 4125 bp, containing four exons and three introns. The ORF encoded a protein of 174 amino acids without a signal sequence. The deduced ferritin contained a highly conserved motif for the ferroxidase center comprising seven residues of a typical vertebrate heavy-chain ferritin. It contained one conserved iron associated residue (Try27) and iron-binding region signature 1 residues. The mRNA contained a 27 bp iron-responsive element with a typical stem-loop structure in the 5'-UTR position. Copy number variants (CNVs) of Fth1 in two populations (PY and JH) were detected using quantitative real-time PCR. Associations between CNVs and growth were also analyzed. The results showed that the copy number of the ferritin gene of in the diploid genome ranged from two to 12 in PY, and from two to six in JH. The copy number variation in PY was higher than that in JH. In terms of shell length, mussels with four copies of the ferritin gene grew faster than those with three copies (P<0.05), suggesting that CNVs in the ferritin gene are associated with growth in shell length and might be a useful molecular marker in selective breeding of H. cumingii.

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