Crassostrea gigas is a frequently studied species in understanding physiological processes in bivalves. Similar to other animals, oysters store glucose in the body as glycogen. Glycogen is known to supply energy for germ cell development and maintenance. Glycogen is synthesized by glycogen synthase. GSK3β regulates glycogen synthase activity and plays an important role in glycogen synthesis. Therefore, this study aims to examine the effect of GSK3β on the annual cycle of oysters and the glycogen synthesis pathway and to investigate the energy pathway in comparison with seasonal variation. Oysters were sampled monthly for one year and were subjected to glycogen content, RT-PCR, FISH, and western blot analysis. The year-round glycogen content significantly differs only in the mantle edge during spring and summer of both sexes but not in labial palp, digestive gland, gonad, and adductor muscle. The expression of GSK3β mRNA level was highest in October for females and April for males. Both sexes had the lowest expression in July. In the adductor muscle, females and males showed the highest expression in April and the lowest in July and October. The pattern of GSK3β expression in gonads and adductor muscle was similarly confirmed through FISH. As a result of examining the signaling system, p-GSK3β (serine 9) increased. At the same time, glycogen synthase decreased in May when the condition index was the highest, p-GSK3β decreased in October and July when spawning occurred, and glycogen synthase increased. Overall, it is thought that p-GSK3β expression is high in C. gigas at ripe, which inhibits glycogen synthesis and is used as energy for growth and maturation. Glycogen synthesis occurs for energy storage during degeneration.

Olive flounder (Paralichthys olivaceus) muscle satellite cells (OFMCs) were obtained by enzymatic primary cell isolation and the explant method. Enzymatic isolation yielded cells that reached 80% confluence within 8 days, compared to 15 days for the explant method. Optimum OFMC growth was observed in 20% fetal bovine serum at 28℃ with NaCl2 and CaCl2, as well as an essential fibroblast growth factor. Olive flounder skeletal muscle myoblasts were induced into a mitogen-poor medium containing 2% horse serum for differentiation; they fused to form multinucleate myotubes. The results indicated complete differentiation of myoblasts into myotubes; we also detected the expression of the myogenic regulatory factors myoD, myogenin, and desmin. Upregulation (Myogenin, desmin) and downregulation (MyoD) of muscle regulation factors confirmed the differentiation in OFMC.