Two Figla homologues have disparate functions during sex differentiation in half-smooth tongue sole (Cynoglossus semilaevis)
Figla is a germ-cell-specific transcription factor associated with ovary development and differentiation. In vertebrates, one transcriptional form of Figla is commonly found. However, besides the common form of this gene (named Figla_tv1), a new transcriptional form (named Figla_tv2) was identified in half-smooth tongue sole (Cynoglossus semilaevis). The full-length cDNA of Figla_tv1 was 1057 bp long with a 591-bp open reading frame encoding a predicted 196 amino acid protein, while Figla_tv2 encoded a 125 amino acid protein. Figla_tv1 and Figla_tv2 expression in various tissues was detected by qRT-PCR. Figla_tv1 was expressed mainly in ovary, skin and liver, while Figla_tv2 was expressed in all examined tissues. In the gonads, Figla_tv1 was expressed in ovary, while Figla_tv2 was predominately expressed in testis of pseudomales. Further, in situ hybridization located Figla_tv1 only in oocytes and Figla_tv2 mainly in germ cells of pseudomale testis. After knocking down Figla_tv2 in a pseudomale testis cell line, the expression of two steroid hormone-encoding genes, StAR and P450scc, was significantly up-regulated (P < 0.05). Our findings suggest that Figla_tv1 has a conserved function in folliculogenesis, as in other vertebrates, and that Figla_tv2 may have a role in the spermatogenesis of pseudomales by regulating the synthesis and metabolism of steroid hormones.
[…] The 5′- and 3′-RACE fragments were assembled using DNASTAR software (DNASTAR, Madison, WI) to obtain the full-length cDNA sequences of Figla_tv1 and Figla_tv2. Sequence identity and similarity of the two translated Figla proteins from C. semilaevis with known Figla proteins from other vertebrates were assessed using the MegAlign program (DNASTAR) with Clustal W method. Sequence alignments of two Figla nucleotide sequences were generated using the Clustal X and GeneDoc programs. Multiple sequence alignments of Figla proteins from different species were conducted using the Clustal X and edited by BOXSHADE (http://www. ch.embnet.org/software/BOX_form.html). […]
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