Production of Pacific abalone progeny from cryopreserved sperm: Molecular insights into growth, reproduction, and immunity

Abstract

This study aimed to produce market-sized Pacific abalone, Haliotis discus hannai progeny from cryopreserved sperm (PCS) and assess their mRNA stability. Embryonic development in PCS was moderately delayed until the veliger larval stage compared to control progeny (CP), although both groups exhibited similar morphological structures. Shell width (SW) and shell height (SH) in PCS and CP remained consistent throughout the experimental periods. However, the shell length (SL) and body weight (BW) of PCS were significantly lower than those of CP during the early developmental stages (up to 9 months). After this period, PCS and CP exhibited comparable body traits at 18 months and 3 years. At 9 months, the mRNA expression patterns of growth-associated genes (Hdh-HGAP, Hdh-IGFBP7, Hdh-MSTN), shell-forming genes (Hdh-PRLSTN, and Hdh-SMP5), immune regulatory genes (Hdh-CAT, Hdh-GR, Hdh-GPx, Hdh-Caspase 3, Hdh-BAX, and Hdh-BCL2) were similar between PCS and CP. However, Hdh-MGF, Hdh-SOD, Hdh-IL17, and Hdh-NF-kB exhibited significantly lower mRNA expression, whereas Hdh-MIRP3 showed significantly higher expression in PCS compared to CP at 9 months. Furthermore, the expression patterns of all examined genes were similar in both PCS and CP at 3 years. In summary, PCS initially demonstrated lower growth traits (SL, BW) and impaired mRNA stability until 9 months; however, these traits normalized over time, resulting in comparable growth traits and gene expression to CP by 3 years. These findings suggest that cryopreserved sperm can effectively produce viable offspring with normal growth traits and mRNA stability for vital growth (Hdh-MGF, Hdh-HGAP, Hdh-IGFBP7, Hdh-MIRP3 and Hdh-MSTN), shell formation (Hdh-SMP5 and Hdh-PRLSTN), reproduction (Hdh-GnRH, Hdh-GnRH-R, Hdh-APGWamide and Hdh-5HT), and immune (Hdh-SOD, Hdh-CAT, Hdh-GR, Hdh-GPx, Hdh-BCL2, Hdh-BAX, Hdh-Caspase 3, Hdh-IL17 and Hdh-NF-kB) associated genes.