Cloning Of Fucal In Monopterus Albus And The Differentiation Expression Of SSEA And Fucal In Monopterus Albus Gonad Tissue

The first mature female Monopterus albus is a hermaphrodite fish. For the Monopterus albus' natural sex reversal phenomenon of the molecular mechanism and genetic background in the academic community has been the focus of attention. To elucidate the developmental and differential situation of female and male germ stem cells (GSCs) in the development and sex reversal of Monopterus Albus, we investigated the location of SSEA1,SSEA4,E-Cadherin (E-Cad) SOX2. Podocalyxin (PCX),OCT3\4 and Nanog in the female and male GSCs. The immunohistochemistry showed that the expression of SSEA-1,SSEA-4,PCX in the male GSCs found more than in the female GSCs which showed that the female GSCs and male GSCs are difference in the biochemical characteristics. It also showed that SSEA in male gonadal development and differentiation in Monopterus Albus may have important biological functions.SSEA is a type of glycosidic lipid, is a secondary product of gene expression. The expression and activity of Fucal (α-L-FucosidaseⅠ) is important to the SSEA synthesis regulation. Use RACE and RT-PCR technology cloned Monopterus albus Fucal partial sequence of gene sequence. Partial cDNA of Monopterus albus Fucal gene long 1176bp, encoding a polypeptide containing a total of 392 amino acids. Sequence alignment analysis showed that Monopterus albus Fucal gene homology between different species of vertebrates are less, suggesting that this gene is not conservative in evolution.Fucal gene showed significant differences in the expression of Monopterus albus Male and female gonadal tissue by RT-PCR. This result is consistent with the results of SSEA in Monopterus albus gonads of immunohistochemistry, which shows that Fucal and SSEA closely related to the catabolism, may be involved in reproductive and developmental regulation of Monopterus albus.In order to further carry out the reversal of Monopterus albus molecular mechanism of reproductive regulation, we explored the methods of the purification, culture and identification of the gonad tissues of Monopterus Albus in vitro. The tissue block pasted culture method were used to culture gonad tissues of Monopterus Albus, the cells migrated out around tissues after 3-5 days culturing and the growth halo emerged. About 15 days, the diameter of the growth halo is up to 3-3.2 mm, the cells of the growth halo include three types:fibroblasts, nerve like cells, and germ stem cell similar cells. Fibroblast was the main components of the growth halo. Its body was flat. The morphology was unregular and there were many short prominences and the light nuclear of cell had 2-3 obvious nucleo luses. The main components of the growth halo. The morphology of Nerve-like cells. Nerve-like cells with radiation filamentous cell protrusions were positive in oil red O staining. Germ stem cell similar cells are round, single-core or multi-core, and the ratio of the nucleus and cytoplasm is mass, AP staining shows positive. Therefore, this study has laid a foundation for the mechanism of sex reversal of Monopterus Albus on the molecule level.