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Molecular Cloning and Functional Characterization of Catalase in Stress Physiology, Innate Immunity, Testicular Development, Metamorphosis, and Cryopreserved Sperm of Pacific Abalone.

ABSTRACT: Catalase is a crucial enzyme of the antioxidant defense system responsible for the maintenance of cellular redox homeostasis. The aim of the present study was to evaluate the molecular regulation of catalase (Hdh-CAT) in stress physiology, innate immunity, testicular development, metamorphosis, and cryopreserved sperm of Pacific abalone. Hdh-CAT gene was cloned from the digestive gland (DG) of Pacific abalone. The 2894 bp sequence of Hdh-CAT had an open reading frame of 1506 bp encoding 501 deduced amino acids. Fluorescence in situ hybridization confirmed Hdh-CAT localization in the digestive tubules of the DG. Hdh-CAT was induced by different types of stress including thermal stress, H₂O₂ induction, and starvation. Immune challenges with Vibrio, lipopolysaccharides, and polyinosinic-polycytidylic acid sodium salt also upregulated Hdh-CAT mRNA expression and catalase activity. Hdh-CAT responded to cadmium induced-toxicity by increasing mRNA expression and catalase activity. Elevated seasonal temperature also altered Hdh-CAT mRNA expression. Hdh-CAT mRNA expression was relatively higher at the trochophore larvae stage of metamorphosis. Cryopreserved sperm showed significantly lower Hdh-CAT mRNA expression levels compared with fresh sperm. Hdh-CAT mRNA expression showed a relationship with the production of ROS. These results suggest that Hdh-CAT might play a role in stress physiology, innate immunity, testicular development, metamorphosis, and sperm cryo-tolerance of Pacific abalone.

SUBMITTER: Hossen S

PROVIDER: S-EPMC9854591 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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Molecular Cloning and Functional Characterization of Catalase in Stress Physiology, Innate Immunity, Testicular Development, Metamorphosis, and Cryopreserved Sperm of Pacific Abalone.

Hossen Shaharior S Sukhan Zahid Parvez ZP Kim Soo Cheol SC Hanif Md Abu MA Kong Il-Keun IK Kho Kang Hee KH

Antioxidants (Basel, Switzerland) 20230101 1

Catalase is a crucial enzyme of the antioxidant defense system responsible for the maintenance of cellular redox homeostasis. The aim of the present study was to evaluate the molecular regulation of catalase (Hdh-CAT) in stress physiology, innate immunity, testicular development, metamorphosis, and cryopreserved sperm of Pacific abalone. Hdh-CAT gene was cloned from the digestive gland (DG) of Pacific abalone. The 2894 bp sequence of Hdh-CAT had an open reading frame of 1506 ...[more]

PMID: 36670971

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Project description:Pacific abalone, Haliotis discus hannai, is a highly commercial seafood in Southeast Asia. The present study aimed to determine the influence of saccharides and vitamins on post-thaw sperm quality, ATP content, fertilization capacity, hatching capacity, and mRNA content of motility and fertilization-associated genes of Pacific abalone. Sperm cryopreserved using saccharides improved the post-thaw sperm quality including motility, acrosome integrity (AI), plasma membrane integrity (PMI), and mitochondrial membrane potential (MMP). However, vitamins (l-ascorbic acid) did not result in any significant improvement in sperm quality. Sperm cryopreserved using saccharides also improved ATP content, DNA integrity, and mRNA content of motility and fertilization-associated genes of post-thaw sperm than sperm cryopreserved without saccharides. Among sperm cryopreserved using different saccharides, post-thaw sperm quality indicators (except PMI) and mRNA content of motility and fertilization-associated genes did not show significant differences between sperm cryopreserved using 3% sucrose (S) combined with 8% dimethyl sulfoxide (DMSO) and sperm cryopreserved using 1% glucose (G) combined with 8% ethylene glycol (EG). However, sperm cryopreserved using 3% S + 8% DMSO showed higher post-thaw sperm quality (motility: 58.4 ± 2.9%, AI: 57.1 ± 3.2%, PMI: 65.3 ± 3.3%, and MMP: 59.1 ± 3.2%), ATP content (48.4 ± 1.8 nmol/ml), and % DNA in tail (2.09 ± 0.20%) than sperm cryopreserved using other saccharides. When sperms were cryopreserved using 3% S + 8% DMSO, the mRNA content of motility (heat shock protein 70, HSP70; heat shock protein 90, HSP90; protein kinase A, PKA-C; axonemal protein 66.0, Axpp66.0; and tektin-4) and fertilization-associated (sperm protein 18 kDa, SP18 kDa) genes were higher than in sperm cryopreserved using other saccharides. However, changes in the mRNA contents of these genes were insignificant between sperm cryopreserved using 3% S + 8% DMSO and 1% G + 8% EG. Taken together, these results indicate that cryopreservation using 3% S + 8% DMSO can improve post-thaw sperm quality and mRNA contents better than other examined cryoprotectants. The present study suggests that 3% S + 8% DMSO is a suitable cryoprotectant for sperm cryopreservation and molecular conservation of this valuable species.

Project description:Cryopreservation of immature testicular tissues is essential for increasing the possibilities of offspring generation by testicular xenografting for agricultural or medical purposes. However, successful production of offspring from the sperm involved has never been reported previously. In the present study, therefore, using intracytoplasmic sperm injection (ICSI), we examined whether xenogeneic sperm obtained from immature pig testicular tissue after cryopreservation would have the capacity to produce live piglets. Testicular fragments from 9- to 11-day-old piglets were vitrified after 10- or 20-min immersion in vitrification solution containing ethylene glycol (EG), polyvinyl pyrrolidone (PVP) and trehalose as cryoprotectants, and then stored in liquid nitrogen for more than 140 days. Thirty nude mice were assigned to each immersion-time group. Testicular fragments were transplanted under the back skin of castrated mice immediately after warming and removal of the cryoprotectants. Blood and testicular grafts were then recovered from the recipient mice on days 60, 120, 180 and 230-350 (day 0?=? grafting). Histological assessment of the testicular grafts and analyses of inhibin and testosterone production revealed no significant differences between the two immersion-time groups, indicating equal growth activity of the cryopreserved tissues. A single sperm obtained from a mouse in each group on day 230-350 was injected into an in vitro-matured porcine oocyte, and then the ICSI oocytes were transferred to the oviducts of estrus-synchronized recipient gilts. One out of 4 gilts that had received oocytes fertilized using sperm from the 10-min immersion group delivered 2 live piglets, and one of another 4 gilts from the 20-min group delivered 4 live piglets. Thus, we have successfully generated porcine offspring utilizing sperm from immature testicular tissues after cryopreservation and transplantation into nude mice. The present model using pigs will be applicable to many large animals, since pigs are phylogenetically distant from the murine recipients.

Dataset Information

Molecular Cloning and Functional Characterization of Catalase in Stress Physiology, Innate Immunity, Testicular Development, Metamorphosis, and Cryopreserved Sperm of Pacific Abalone.

Publications

Molecular Cloning and Functional Characterization of Catalase in Stress Physiology, Innate Immunity, Testicular Development, Metamorphosis, and Cryopreserved Sperm of Pacific Abalone.

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