Structural and expression analyses of gonadotropin Ibeta subunit genes in goldfish (Carassius auratus)

Gonadotropin (GTH) is a pituitary glycoprotein hormone that regulates gonadal development in vertebrates. In teleosts, it is considered that two types of GTH, GTH I (follicle-stimulating hormone-like GTH) and GTH II (luteinizing hormone-like GTH), are produced in the pituitary, and their molecules are comprised of common alpha and distinct beta subunits. In this study, we describe the complete structure and 5'-flanking regulatory region of two distinct genes encoding GTH Ibeta in goldfish, Carassius auratus. The two goldfish GTH Ibeta genes, gfGTHIbeta-1 and gfGTHIbeta-2, span 1719 and 1545 base pairs (bp) nucleotides, respectively, and there is a high sequence identity (92.1%) between the coding regions. Both genes consist of three exons separated by two introns as in mammalian FSH beta genes. The locations of the first intron and second intron showed a well-conserved pattern similar to those of mammalian FSH beta genes. Inspection of the 5'-flanking region of the gfGTHIbeta-1 and gfGTHIbeta-2 (approximately 1.4 and 1.1kb, respectively) revealed the presence of several putative cis-acting elements, including the gonadotrope-specific element, gonadotropin-releasing hormone responsive element, and half steroid hormone responsive elements. Interestingly, some of their elements were located contiguously between -187 and -124bp upstream from a TATAA sequence. Reverse transcription polymerase chain reaction confirmed that these two genes are expressed in the pituitary of individual fish. These results, taken together, demonstrate that there are at least two functional genes encoding GTH Ibeta, probably due to the tetraploidy of goldfish. The unique locations of the cis-acting elements in the GTH Ibeta genes suggest they may be involved in the expression of the goldfish GTH Ibeta gene.