Multiple vitellogenin genes (vtgs) in large yellow croaker (Larimichthys crocea): molecular characterization and expression pattern analysis during ovarian development.

Abstract

The large yellow croaker (Larimichthys crocea) is a marine fish that is economically important to Chinese fisheries, and its reproductive and developmental biology have been extensively investigated. However, the molecular mechanism of oogenesis in L. crocea is not clear. Here, we investigated the multiple vitellogenin (Vtg) system in large yellow croaker. Three different vtg cDNA sequences, including vtgAa, vtgAb and vtgC, were cloned, which indicate the existence of multiple Vtg proteins in large yellow croaker (Lc-Vtgs). Subsequently, the vtg cDNA sequences and predicted Vtg protein structures were analysed, and Vtg protein structures were found to be highly conserved. To research the expression of vtgs during the development of the ovaries, we examined ovarian development and oogenesis by histological analysis. Four stages of ovary development - stages II, III, IV and V - were observed and their boundaries were defined. Soon afterwards, the expression of vtgs in the liver (known as the main site of Vtg synthesis in teleosts) and ovary were analysed. The expression of vtgs was detected in the two tissues. Interestingly, in the early stages of development (stages II and III), there is little or no generation of yolk granules and the expression of vtgs in the liver is low. However, in the late stages (stages IV and V), yolk granules are generated rapidly and the expression of vtgs is significantly increased in the liver. These results support the hypothesis that the Vtgs were synthetized by the liver, and absorbed by the growing oocytes to promote oogenesis in large yellow croaker. We also detected the presence of vtg mRNA in the liver cells and oocytes by in situ hybridization, which indicated that vths were expressed both in the liver and ovaries. Importantly, we found that the distribution of vtgAa and vtgAb mRNA was close to the sites of yolk granule formation in oocytes.