Differential expression of Gnrh2, Gthbeta, and Gthr genes in sterile triploids and fertile tetraploids.
ABSTRACT Gonadotropin-releasing hormone (GnRH), gonadotropin hormone (GTH), and gonadotropin hormone receptor (GTHR) are the pivotal signal molecules of the hypothalamic-pituitary-gonad (HPG) axis, which plays a crucial role in regulating gonadal development in vertebrate. In this study, we comparatively analyze the expression characteristics of Gnrh2, Gthbeta, and Gthr in red crucian carp diploids, triploids, and allotetraploids. The expression patterns of these genes are similar in the three fish ploidy types: the Gnrh2 gene is expressed in midbrains, pituitaries, and gonads; the Gthbeta gene is expressed in pituitaries; the Gthr gene is mainly expressed in gonads. These results indicate that the three genes participate in the regulation of gonadal development. By real-time polymerase chain reaction and in situ hybridization, we find that, among these three fish ploidy types, the expression level of Gthr in the gonads of triploids is lower than those of diploids and tetraploids; this weakens the combination of GTHR with GTH released from the pituitary and leads to the sterility of triploids, since the gonad cannot produce enough sex steroids. In addition, the low expression of Gthr in triploids may affect the down-regulation of Gthbeta, which then affects the down-regulation of Gnrh2; hence, the expression levels of Gnrh2 and Gthbeta genes in triploids are the highest after the breeding season. In conclusion, the differential expression of Gnrh2, Gthbeta, and Gthr in triploids and tetraploids is related to their sterility and bisexual fertility, respectively.
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ABSTRACT: Increasing piRNAs provide RNA-interference pathways to regulate transposons and gene expression in germline cells. We demonstrate that Piwi transcripts are exclusively expressed in adult testes and ovaries in teleosts, with triploids showing the highest Piwi expression in the ovaries. Studies in vivo and in vitro showed that hCG and E2 treatment suppressed Piwi expression. We further cloned 200 small RNAs in the three kinds of fish. Seven piRNAs were obtained from all the three different ploidy fishes. During ovulation, five piRNAs showed significantly higher expression in the ovaries of sterile triploids than fertile diploids and tetraploids. Furthermore, E2 suppressed the expression of the six piRNAs at different levels in vivo and in vitro. The present study bridges the gap between the HPG axis and Piwi-piRNA pathway by suggesting that a dysfunctional HPG axis abrogated the piRNA suppression in triploid fish.Molecular and Cellular Endocrinology 12/2013; · 4.04 Impact Factor
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ABSTRACT: Distant hybridization makes it possible to transfer the genome of one species to another, which results in changes in phenotypes and genotypes of the progenies. This study shows that distant hybridization or the combination of this method with gynogenesis or androgenesis lead to different ploidy fishes with genetic variation, including fertile tetraploid hybrids, sterile triploid hybrids, fertile diploid hybrids, fertile diploid gynogenetic fish, and their derived progenies. The formations of the different ploidy fishes depend on the genetic relationship between the parents. In this study, several types of distant hybridization, including red crucian carp (Carassius auratus red var.) (2n=100, abbreviated as RCC) (female) x common carp (Cyprinus carpio L.) (2n=100, abbreviated as CC) (male), and RCC (2n=100) (female) x blunt snout bream (Megalobrama amblycephala) (2n=48, abbreviated as BSB) (male) are described. In the distant hybridization of RCC (female) x CC (male), bisexual fertile F(3)-F(18) allotetraploid hybrids (4n=200, abbreviated as 4nAT) were formed. The diploid hybrid eggs and diploid sperm generated by the females and males of 4nAT developed into diploid gynogenetic hybrids and diploid androgenetic hybrids, respectively, by gynogenesis and androgenesis, without treatment for doubling the chromosome. Improved tetraploid hybrids and improved diploid fishes with genetic variation were derived from the gynogenetic hybrid line. The improved diploid fishes included the high-body RCC and high-body goldfish. The formation of the tetraploid hybrids was related to the occurrence of unreduced gametes generated from the diploid hybrids, which involved in premeiotic endoreduplication, endomitosis, or fusion of germ cells. The sterile triploid hybrids (3n=150) were produced on a large scale by crossing the males of tetraploid hybrids with females of diploid fish (2n=100). In another distant hybridization of RCC (female) x BSB (male), different ploidy fishes were obtained, including diploid bisexual fertile natural gynogenetic fish (2n=100), sterile triploid hybrids (3n=124), and bisexual fertile tetraploid hybrids (4n=148). Furthermore, two kinds of pentaploid hybrids (5n=172 and 5n=198) were formed. The biological characteristics and the mechanisms of formation of the different ploidy fish were compared and discussed at the cellular and molecular level. The results indicated distant hybridization or the combination of this method with gynogenesis or androgenesis affects the formation of different ploidy fish with genetic variation.Science China. Life sciences 04/2010; 53(4):416-25. · 2.02 Impact Factor
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ABSTRACT: In vertebrates, gonadotropin-releasing hormone (GnRH) and pituitary adenylate cyclase-activating polypeptide (PACAP) are key hormones regulating growth and reproduction in the brain-pituitary axis. The regulating hormonal interactions are of great interest, therefore, the aim of this study is to provide novel insights into the involvement of brain GnRH and PACAP in oogensis and spermatogenesis in a fish model, the blue gourami (Trichogaster trichopterus). cDNA cloning of two GnRH forms combined with phylogenetic analysis revealed that three paralogous GnRH forms exist in blue gourami and evolve as a result of genome duplication. GnRH1 mRNA levels are related to final oocyte maturation (FOM), and this peptide stimulated β follicle-stimulating hormone (βFSH) and growth hormone (GH) gene expression; GnRH2 stimulated β gonadotropins (GtH) gene expression and GnRH analog combined with PACAP-38 synergistically upregulate GH and βFSH gene expression. The data presented, together with previous studies in our lab, enable suggesting mechanisms explaining the physiological relevance of these peptides in the regulation of gametogenesis and steroidogenesis in blue gourami females. These findings support the biological importance of the GnRH and PACAP hormones family, enabling them to stimulate differential biological functions in the regulation of growth and reproduction.Journal of Molecular Neuroscience 03/2012; 48(3):603-16. · 2.89 Impact Factor