Project description:As a precise and efficient genetic engineering method, gene editing technology for animals and plants will gradually move towards large-scale applications, and will have a disruptive impact on the breeding of related traits. By using antisense RNA interference technology to interfere with the mRNA expression and protein translation of the SF-1 gene, we have successfully cultivated tilapia with defective gonadal development. The results of the study showed that the weights of male and female fish in the experimental group were significantly higher than those of the control group. The gonadal weight and gonadal index of the experimental group were significantly lower than those of the control group and the negative control group. HE staining showed that the number of spermatogonia in the testis tissue of the experimental group was significantly reduced, the cavity in the seminiferous tubules was significantly enlarged and increased, the number of sperm cells was significantly decreased, and the number of sperm was relatively rare. However, a large number of stage V oocytes accumulated in the ovarian tissue of female fish in the experimental group, and many of them were gray-white. The mRNA and protein expression levels of SF-1 gene in the gonadal tissues were extremely significantly lower than those of the control and negative controls. The combined analysis of transcriptome and proteome showed that Steroid hormone biosynthesis, Arachidonic acid metabolism and Cell adhesion molecules are involved in the regulation of the gonads of male fish after SF-1 gene silencing. However, ECM-receptor interaction, Retinol metabolism, Nicotinate and nicotinamide metabolism and cytochrome P450 may be involved in the regulation of female fish SF-1 gene silencing. SF-1 silencing may cause developmental defects in the external reproductive organs of male and female fishes and inhibit the development of gonadal tissues through different regulatory pathways. This provides data support for the further functional analysis of SF-1 on fish and the treatment of abnormal human SF-1 expression.

Project description:Background: Food supply is a major factor influencing growth rates in animals. This has important implications for both natural and farmed fish populations, since food restriction may difficult reproduction. However, a study on the effects of food supply on the development of juvenile gonads has never been transcriptionally described in fish. Methods and Findings: This study investigated the consequences of growth on gonadal transcriptome of European sea bass in: 1) 4-month-old sexually undifferentiated fish, comparing the gonads of fish with the highest vs. the lowest growth, to explore a possible link between transcriptome and future sex, and 2) testis from 11-month-old juveniles where growth had been manipulated through changes in food supply. The four groups used were: i) sustained fast growth, ii) sustained slow growth, iii) accelerated growth, iv) decelerated growth. The transcriptome of undifferentiated gonads was not drastically affected by initial natural differences in growth. Further, changes in the expression of genes associated with protein turnover were seen, favoring catabolism in slow-growing fish and anabolism in fast-growing fish. Moreover, while fast-growing fish took energy from glucose, as deduced from the pathways affected and the analysis of protein-protein interactions examined, in slow-growing fish lipid metabolism and gluconeogenesis was favored. Interestingly, the highest transcriptomic differences were found when forcing initially fast-growing fish to decelerate their growth, while accelerating growth of initially slow-growing fish resulted in full transcriptomic convergence with sustained fast-growing fish. Conclusions: Food availability during sex differentiation shapes the juvenile testis transcriptome, as evidenced by adaptations to different energy balances. Remarkably, this occurs in absence of major histological changes in the testis. Thus, fish are able to recover transcriptionally their testes if they are provided with enough food supply during sex differentiation; however, an initial fast growth does not represent any advantage in terms of transcriptional fitness if later food becomes scarce.

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