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: Triploid carp (100%) with 150 (3n=150) chromosomes were obtained by crossing the females of improved tetraploid hybrids (♀, 4n=200) of red crucian carp (♀)×common carp (♂) with the males of diploid yellow river carp (♂, 2n=100). The crosses yielded transgenic triploid carp (positive triploid fish, 44.2% of the progeny) and non-transgenic triploid carp (negative triploid fish). Histological examination of the gonads of 24-month-old positive triploid fish suggested they were sterile and the fish were not able to produce mature gametes during the breeding season. Morphologically, both the positive and negative triploid fish were similar. They had a spindle-shaped, laterally compressed, steel grey body with two pairs of barbells. Most of the quantifiable traits of the triploid carp were intermediate between those of the two parents. The positive and negative triploid fish were raised in the same pond for 2 years. The mean body weight of the positive triploid fish was 2.3 times higher than the negative triploid fish. The weight of the largest positive triploid fish was 2.91 times higher than that of the largest negative triploid fish. Thus, we produced fast-growing transgenic triploid carp that have a reduced ecological risk because of their inability to mate and produce progeny. Keywordstransgene–carp–diploid–triploid–improved tetraploid hybrids of red crucian carp×common carp–sterility–ecological safetyChinese Science Bulletin 06/2011; 56(16):1679-1684. · 1.37 Impact Factor
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ABSTRACT: In this study, the annual cycle of the gonadal steroids testosterone (T), 11-ketotestosterone (11-KT), 17β-estradiol (E2) and 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) was determined using radioimmunoassay and then compared for two populations of rainbow trout, XX diploid females (n = 40) and XXX triploid females (n = 15). In females, E2 and DHP levels were found to be significantly related to body weight (r = 0.22513; p < 0.0001 and r = 0.15831; p > 0.001, respectively). In this group, E2 concentrations peaked in November (25.05 ng /ml), while maximum DHP levels, only measurable from October to April, were attained in February (64.14 ng /ml). No significant differences in hormone ranges related to egg output ability were observed. Finally, sex steroid concentrations were low in the triploid female XXX fish compared to the female XX population. Nevertheless, maximum T (33.85 ng /ml) and 11-KT (32.35 ng /ml) levels were recorded in January, for XXX. The levels for these two hormones are relatively high and are also significantly associated (r = 0.8430; p < 0.0001). Diploid females showed significantly higher levels of E2 than triploids over the 12-month study period. The female triploid fish produced the lowest steroid hormone levels, such that these would be the most suitable for human consumption.Reproduction in Domestic Animals 06/2012; · 1.18 Impact Factor
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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; · 4.24 Impact Factor