The factor in the germline alpha(figla), as a member of the basic helix-loop-helix family, has been reported to be involved in ovary development in mammals and teleosts. However, the regulatory mechanisms of figla in ...The factor in the germline alpha(figla), as a member of the basic helix-loop-helix family, has been reported to be involved in ovary development in mammals and teleosts. However, the regulatory mechanisms of figla in teleosts remain unclear. Here,figla in P. olivaceus(Pofigla) was characterized with encoding a 202 amino acid protein that contains a conserved basic region and helix-loop-helix(HLH) domain. Amino acids alignment and synteny analysis revealed that Pofigla was conserved with the orthologous gene sequences in other vertebrates. The results of qRT-PCR showed Pofigla was maternally inherited during embryonic development. For tissue distribution, Pofigla showed a sexually dimorphic gene expression in the gonad of different genders, with a higher expression in ovary than in testis. In situ hybridization(ISH) results demonstrated Pofigla was specifically expressed in germ cells including oocytes, spermatogonia and spermatocytes. By screening and analyzing two proximal regions(-2966/-2126 and-772/-444) with high promoter activity, we found SOX5, LEF1, FOXP1 and GATA1 may play important roles in the transcriptional regulation of Pofigla. Furthermore, we observed the co-localization between Figla and LEF1 in HEK 293T cells. And the significant up-regulation effect of the canonical Wnt signaling pathway on the expression of Pofigla was found in cultured ovarian cells. This study provided the first evidence that figla not only has an important function in ovary development, but also plays some potential roles in testis development and/or male germ cell differentiation during early testis development in P. olivaceus. The results provide valuable reference in exploring the regulatory network of figla in teleost.展开更多
MiR-430 is a non-coding small RNA that participates in early embryonic development by acting on the Nodal signaling pathway.Its functions in Paralichthys olivaceus(P.olivaceus),however,remain unclear.In this study,thr...MiR-430 is a non-coding small RNA that participates in early embryonic development by acting on the Nodal signaling pathway.Its functions in Paralichthys olivaceus(P.olivaceus),however,remain unclear.In this study,through biological information technology,we hypothesized lefty was the target gene of miR-430.Lefty is a divergent member of the transforming growth factor-β(TGF-β)superfamily,and is involved in regulating left-right patterning as an antagonist of the Nodal signaling pathway.To study the relationship between miR-430 and lefty gene,miR-430 and fluorescent expression vector were transfected into embryo cells.Our results showed that miR-430 has important effects on early embryonic development.It can inhibit the expressions of lefty and other genes related to Nodal signaling pathway.Additionally,the expression of miR-430 was involved in the metamorphosis of P.olivaceus.This study clarifies the function of miR-430 in P.olivaceus,which will be helpful for further research on the interactions between miR-430 and the Nodal signaling pathway.展开更多
基金supported by the National Key R&D Program of China (No. 2018YFD0901205)
文摘The factor in the germline alpha(figla), as a member of the basic helix-loop-helix family, has been reported to be involved in ovary development in mammals and teleosts. However, the regulatory mechanisms of figla in teleosts remain unclear. Here,figla in P. olivaceus(Pofigla) was characterized with encoding a 202 amino acid protein that contains a conserved basic region and helix-loop-helix(HLH) domain. Amino acids alignment and synteny analysis revealed that Pofigla was conserved with the orthologous gene sequences in other vertebrates. The results of qRT-PCR showed Pofigla was maternally inherited during embryonic development. For tissue distribution, Pofigla showed a sexually dimorphic gene expression in the gonad of different genders, with a higher expression in ovary than in testis. In situ hybridization(ISH) results demonstrated Pofigla was specifically expressed in germ cells including oocytes, spermatogonia and spermatocytes. By screening and analyzing two proximal regions(-2966/-2126 and-772/-444) with high promoter activity, we found SOX5, LEF1, FOXP1 and GATA1 may play important roles in the transcriptional regulation of Pofigla. Furthermore, we observed the co-localization between Figla and LEF1 in HEK 293T cells. And the significant up-regulation effect of the canonical Wnt signaling pathway on the expression of Pofigla was found in cultured ovarian cells. This study provided the first evidence that figla not only has an important function in ovary development, but also plays some potential roles in testis development and/or male germ cell differentiation during early testis development in P. olivaceus. The results provide valuable reference in exploring the regulatory network of figla in teleost.
基金supported by the National Natural Sci-ence Foundation of China(No.31372511)the Fun-damental Research Funds for the Central Universities(No.201822026)
文摘MiR-430 is a non-coding small RNA that participates in early embryonic development by acting on the Nodal signaling pathway.Its functions in Paralichthys olivaceus(P.olivaceus),however,remain unclear.In this study,through biological information technology,we hypothesized lefty was the target gene of miR-430.Lefty is a divergent member of the transforming growth factor-β(TGF-β)superfamily,and is involved in regulating left-right patterning as an antagonist of the Nodal signaling pathway.To study the relationship between miR-430 and lefty gene,miR-430 and fluorescent expression vector were transfected into embryo cells.Our results showed that miR-430 has important effects on early embryonic development.It can inhibit the expressions of lefty and other genes related to Nodal signaling pathway.Additionally,the expression of miR-430 was involved in the metamorphosis of P.olivaceus.This study clarifies the function of miR-430 in P.olivaceus,which will be helpful for further research on the interactions between miR-430 and the Nodal signaling pathway.
