Carbonic Anhydrase in Pacific Abalone Haliotis discus hannai: Characterization, Expression, and Role in Biomineralization.

Carbonic anhydrases (CAs) are universal zinc ion containing metalloenzymes that play a pivotal role in various physiological processes. In this study, a CA I (designated as Hdh CA I) was isolated and characterized from the mantle tissue of Pacific abalone, Haliotis discus hannai. The full-length cDNA sequence of Hdh CA I was 1,417-bp in length, encoding a protein of 337 amino acids with molecular weight of 37.58 kDa. Hdh CA I sequence possessed a putative signal peptide of 22 amino acids and a CA catalytic function domain. The predicted protein shared 94 and 78% sequence identities with Haliotis gigantea and Haliotis tuberculata CA I, respectively. Results of phylogenetic analysis indicated that Hdh CA I was evolutionarily close to CA I of H. gigantea and H. tuberculata with high bootstrap values. Significantly higher levels of Hdh CA I mRNA transcript were found in mantle than other examined tissues. In situ hybridization results showed strong hybridization signals in epithelial cells of the dorsal mantle pallial, an area known to synthesize and secrete proteins responsible for the nacreous layer formation of shell. This is the first study on Hdh CA I in H. discus hannai and the results may contribute to further study its physiological functions in shell biomineralization of abalone.

Sharker MR ，Kim SC ，Hossen S ，Sumi KR ，Choi SK ，Choi KS ，Kho KH ... -  《Frontiers in Molecular Biosciences》

Molecular Characterization of Carbonic Anhydrase II (CA II) and Its Potential Involvement in Regulating Shell Formation in the Pacific Abalone, Haliotis discus hannai.

Carbonic anhydrases (CAs) are a family of metalloenzymes that can catalyze the reversible interconversion of CO2/HCO3 -, ubiquitously present in both prokaryotes and eukaryotes. In the present study, a CA II (designated as HdhCA II) was sequenced and characterized from the mantle tissue of the Pacific abalone. The complete sequence of HdhCA II was 1,169 bp, encoding a polypeptide of 349 amino acids with a NH2-terminal signal peptide and a CA architectural domain. The predicted protein shared 98.57% and 68.59% sequence identities with CA II of Haliotis gigantea and Haliotis tuberculata, respectively. Two putative N-linked glycosylation motifs and two cysteine residues could potentially form intramolecular disulfide bond present in HdhCA II. The phylogenetic analysis indicated that HdhCA II was placed in a gastropod clade and robustly clustered with CA II of H. gigantea and H. tuberculata. The highest level of HdhCA II mRNA expression was detected in the shell forming mantle tissue. During ontogenesis, the mRNA of HdhCA II was detected in all stages, with larval shell formation stage showing the highest expression level. The in situ hybridization results detected the HdhCA II mRNA expression in the epithelial cells of the dorsal mantle pallial, an area known to express genes involved in the formation of a nacreous layer in the shell. This is the first report of HdhCA II in the Pacific abalone, and the results of this study indicate that this gene might play a role in the shell formation of abalone.

Sharker MR ，Sukhan ZP ，Sumi KR ，Choi SK ，Choi KS ，Kho KH ... -  《-》

Characterization of Insulin-Like Growth Factor Binding Protein 7 (Igfbp7) and Its Potential Involvement in Shell Formation and Metamorphosis of Pacific Abalone, Haliotis discus hannai.

Sharker MR ，Hossen S ，Nou IS ，Kho KH ... -  《INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES》

Characterization of Insulin-Like Growth Factor Binding Protein-5 (IGFBP-5) Gene and Its Potential Roles in Ontogenesis in the Pacific Abalone, Haliotis discus hannai.

Sharker MR ，Kim SC ，Hossen S ，Kho KH ... -  《Biology-Basel》

Hdh-Tektin-4 Regulates Motility of Fresh and Cryopreserved Sperm in Pacific Abalone, Haliotis discus hannai.

As structural components of sperm, tektins are thought to play a fundamental role in sperm flagellar motility. In this study, Tektin-4 (Hdh-TEKT4) gene was successfully cloned and characterized from the testis tissue in Pacific abalone, Haliotis discus hannai. The full-length cDNA of Hdh-TEKT4 was 1,983 bp, with a coding region of 1,350 bp encoding 51.83 kDa putative protein of 449 deduced amino acids. Hdh-TEKT4 contains a tektin domain including a nonapeptide signature motif (RPGVDLCRD). Fluorescence in situ hybridization revealed that Hdh-TEKT4 localized in the spermatids of Pacific abalone testis. qRT-PCR analysis showed that Hdh-TEKT4 was predominantly expressed in testis tissues. Hdh-TEKT4 mRNA expression was upregulated during the fully mature testicular developmental stage in both seasonal development and EAT exposed abalone. Furthermore, mRNA expression of Hdh-TEKT4 was significantly higher in sperm with higher motility than in sperm with lower motility during peak breeding season, induced spawning activity stages, and after cryopreservation in different cryoprotectants. Taken together, these results indicate that the expression of Hdh-TEKT4 in Pacific abalone sperm might have a positive correlation with sperm motility.

Sukhan ZP ，Hossen S ，Cho Y ，Lee WK ，Kho KH ... -  《Frontiers in Cell and Developmental Biology》