| This study was focused on the wild-type zebrafish male and female morphology,expression of melanin inhibitor on zebrafish embryonic development and the formation of melanin and the innate immune related genes α2ML. The main findings are as follows:(1)Established the equation to identify male and female zebrafish by using statistical methods:Female: S1=335.868X1+206.572X2-48.174Male: S2=388.843X1+155.848X2-44.586The correlation of female zebrafish body weight and body length was greater,compared to the male,the characteristic of female bodily form was higher,the head was lower.(2)The research finding of the effect of PTC on zebrafish: ① The function of zebrafish heart will be decreased with increasing concentration of PTC. ② The juvenile of zebrafish was more tolerant to PTC than the embryonic, and the early zebrafish embryos were more sensitive to PTC. ③Egg membrane has a very strong protective effect on zebrafish embryos. In later experiments, the zebrafish embryo toxicity experiment can be divided into two phases: First, the rupture of membranes before hatching, second, second,the rupture of membranes after hatching. Or you can try to peel eggs directly in the appropriate culture medium for experimental. ④ In this study, the group of 0.3m M concentration has the best melanin inhibiting effect. ⑤The experiment of dark group showed that zebrafish in a dark environment will form more melanin. Suggestions in the light related experiments can be appropriately increase the light.(3)Due to α2ML in gene function during embryonic development was not yet clear. so we chose the Zebrafish as a research subject, using RT-PCR and in situ hybridization techniques to analyze the gene expression patterns of α2ML in fish tissues and embryos of zebrafish. By the result of RT-PCR, the expression of α2ML was detected in the adult liver,intestines, heart, muscle, ovary, testis and gill.Express quantity order: liver > intestinal >muscle > testis > heart > gill > ovaries. In embryos RT-PCR experiments, the expression ofα2ML was only detected in the gastrula stage, pharyngeal gill period and the incubation period. Express quantity order: pharyngeal gill stage> incubation period> gastrulation. It showed cleavage stage of 1h and 2.5h can find a clear signal of expression in theblastoderm. Blastula stage in 3h, 4h, 5.3h, 5.7h, the entire embryonic cell will be visible seen the signal expression of α2ML. Blastula stage in 5.3h and 5.7h,It had obvious yolk fusion nucleus(YSN) expression.It turned out that α2ML may play a role in migration of embryonic cells. Gastrulation period of 6h, 6.5h, 7h, 7.5h, 8h, 9h embryonic cells can be seen the signal of α2ML expression throughout the embryonic cell and both can be seen clearly YSN.Segment generated period in 14 h and 18 h, the signal in germ layers of the embryo is not obvious and transparent,but the embryonic cells on egg yolk and YSN expressed significantly. During the hatching period(60h, 72h), tail epidermis and abdomen yolk in embryo are evident expressed,the liver and intestinal in 72 h express clearly,the retina can also detect the signal. In the juvenile stage of 120 h, It had a strong external signal performance in back, tail, belly and pectoral skin. There was also a strong signal in internal swim bladder, liver, intestine and retina. |
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