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Histology of female and male gonads. (a) Female ovaries at previtellogenic stage or primary growth stage showed chromatin nucleolar oocyte (1) and perinucleolar oocyte (2) previtologenic. (b) Male testis in pre-spermatogenic stage; the testes observed with numerous lobules contain only spermatogonia (1) separated by a thin layer of connective tissue (arrow)
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A study was conducted to evaluate the gonad differentiation of juvenile yellow perch (YP, Perca flavencens) and determine the latest labile period related to hormone treatment. Juvenile fish were subjected to two dietary concentrations of methyltestosterone (MT; 20 and 50 mg/kg feed) for 60 days in three (3) age groups of 38-, 46-, and 67-days post...
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... The MT feeding was administered from 20 to 68 dpb stage, and after that, the feeding of the fish continued on the control diet without the hormone. Three breedingenvironment replicates of each treatment group were set up, and MT concentrations were based on previous studies [16][17][18]. After the 140 dpb stage period, body length, total length, and weight of each individual were measured, and fin, whole-body tissue, and gonadal tissue samples were collected for subsequent hormone-level analysis histological, sex determination, and transcriptomic analyses. ...
It is widely known that all-female fish production holds economic value for aquaculture. Sebastes schlegelii, a preeminent economic species, exhibits a sex dimorphism, with females surpassing males in growth. In this regard, achieving all-female black rockfish production could significantly enhance breeding profitability. In this study, we utilized the widely used male sex-regulating hormone, 17α-methyltestosterone (MT) at three different concentrations (20, 40, and 60 ppm), to produce pseudomales of S. schlegelii for subsequent all-female offspring breeding. Long-term MT administration severely inhibits the growth of S. schlegelii, while short term had no significant impact. Histological analysis confirmed sex reversal at all MT concentrations; however, both medium and higher MT concentrations impaired testis development. MT also influenced sex steroid hormone levels in pseudomales, suppressing E2 while increasing T and 11-KT levels. In addition, a transcriptome analysis revealed that MT down-regulated ovarian-related genes (cyp19a1a and foxl2) while up-regulating male-related genes (amh) in pseudomales. Furthermore, MT modulated the TGF-β signaling and steroid hormone biosynthesis pathways, indicating its crucial role in S. schlegelii sex differentiation. Therefore, the current study provides a method for achieving sexual reversal using MT in S. schlegelii and offers an initial insight into the underlying mechanism of sexual reversal in this species.
... During the study period mortality of the fry was observed in control group T 0 and treatment groups H 1 , H 2 and H 3 respectively (Fig.3). MT was fed to fish at dosages close to the lowest concentration employed in previous research, particularly with cold water fishes (Othman et al, 2022;Farias et al, 2023), they suggested that MT generally boosted growth. The amount of food given with hormone or how long it was given to the fish could be contributing factors in the current study's decreased growth rate (Liu et al, 2023), and certain salmonids. ...
... The transition to PF or PM was induced at the embryonic stage 15th day before sex differentiation by estradiol (E2) or methyltestosterone (MT). We injected 5 µL of 10 mg/mL E2 (E110147, Aladdin, shanghai, China) or MT (M830053, Macklin, shanghai, China) into the fertilized eggs whose eggshells were softened by hydrochloric acid (HCl) to induce the sex reversal of Chinese soft-shelled turtle (Zhou et al., 2021;Othman et al., 2022). The gonads of pseudofemale and pseudo-male individuals began to differentiate at the embryonic stage 15th day and mature after 2 years. ...
Sex dimorphism is a key feature of Chinese soft-shelled turtle ( Pelodiscus sinensis ). The males (M) have higher econosmic value than females (F) due to wider calipash and faster growth. Exogenous hormones like estradiol and methyltestosterone can induce sexual reversal to form new phenotypes (pseudo-female, PF; pseudo-male, PM) without changing the genotype. The possibility of inducing sexual reversal is particularly important in aquaculture breeding, but the underlying biological mechanisms remain unclear. Here we applied a direct RNA sequencing method with ultralong reads using Oxford Nanopore Technologies to study the transcriptome complexity in P. sinensis . Nanopore sequencing of the four gender types (M, F, PF, and PM) showed that the distribution of read length and gene expression was more similar between same-sex phenotypes than same-sex genotypes. Compared to turtles with an M phenotype, alternative splicing was more pronounced in F turtles, especially at alternative 3′ splice sites, alternative 5′ splice sites, and alternative first exons. Furthermore, the two RNA methylation modifications m5C and m6A were differentially distributed across gender phenotypes, with the M type having more modification sites in coding sequence regions, but fewer modification sites in 3′UTR regions. Quantitative analysis of enriched m6A RNAs revealed that the N6-methylated levels of Odf2 , Pacs2 , and Ak1 were significantly higher in M phenotype individuals, while the N6-methylated levels of Ube2o were reduced after sexual reversal from both M and F phenotypes. Taken together, these findings reveal an important role of epigenetics during sexual reversal in Chinese soft-shelled turtles.
To investigate the effects of exogenous steroid hormones on growth, body color, and gonadal development in the Opsariichthys bidens (O. bidens), synthetic methyltestosterone (MT) and 17β-estradiol (E2) were used for 28 days’ treatment of 4-month-old O. bidens before the breeding season. Our results suggested that MT had a significant growth-promoting effect (P < 0.05), whereas E2 played an inhibitory role. On the body surface, the females in the MT group showed gray stripes, and the fish in other groups showed no obvious stripes. The males with MT treatment displayed brighter blue-green stripes compared to the CK and E2 groups. The histological analysis showed that the MT significantly promoted testes development in males, blocked oocyte development, and caused massive apoptosis in females, whereas the E2 group promoted ovarian development and inhibited testes development. Based on qRT-PCR analysis, in females, the expression of igf-1, dmrt1, and cyp19a1a genes revealed that E2 treatment resulted in down-regulation of igf-1 expression and up-regulation of cyp19a1a expression. In males, igf-1 and dmrt1 were significantly up-regulated after MT treatment, and E2 treatment led to down-regulation of igf-1. Therefore, this study demonstrates that MT and E2 play an important role in reversing the morphological sex characteristics of females and males.
The objective of this study was to reveal the growth, colouration and gonado-physiological changes due to the exogenous aromatase inhibitor (AIs) in an ornamental fish. 17α-methyl-testosterone (MT) and letrozole (LET) were used as potential AIs. The AI were supplemented with a gel-based feed (LET: 50, 100, 150 and MT: 12.5, 25, 37.5 mg/kg feed) in Rosy barb, Pethia conchonius fry. The fishes were reared in a 45-L glass tank using AI treated gel-based feed for 3 months. Growth in AI-based diets was reduced but the reduction was minimal compared to the control. At 25 mg/kg feed of 17 MT, the highest male proportion (84.72% 6.05%) was recorded, which was significantly higher (P�0.05) than other groups. L*, a*, and b* values showed that 17α-MT-fed groups had brighter coloration (P�0.05). Histological sections showed that LET-17α-MT suppressed ovarian development , causing atretic oocytes. Testicular development was unaffected. 25 mg/kg-treated feed increased SOD, CAT, GST, and GPX. The AI (MT) at 25 mg/kg gel-based feed could therefore be utilised for musculinization without impacting growth, colour, and antioxidant activity of rosy barb, which serves the entire male population in the ornamental fish sector.
Spats1 (spermatogenesis-associated, serinerich 1) has been characterized as a male-biased gene which acts an important role in the germ cell differentiation of mammals. Nevertheless, the function of Spats1 in the Chinese soft-shelled turtle (P. sinensis) has not yet been reported. To initially explore the expression of Spats1 in P. sinensis and its response to sex steroid treatment, we cloned the CDS of Spats1 for the first time and analyzed its expression profile in different tissues, including the testes in different seasons. The Spats1 cDNA fragment is 1201 base pairs (bp) in length and contains an open reading frame (ORF) of 849 bp, which codes for 283 amino acids. Spats1 mRNA was highly expressed in the testes (p < 0.01) and barely detectable in other tissues. In P. sinensis, the relative expression of Spats1 also responsive to seasonal changes in testis development. In summer (July) and autumn (October), Spats1 gene expression was significantly higher in the testes than in other seasons (p < 0.05). Spats1 mRNA was found to be specifically expressed in germ cells by chemical in situ hybridization (CISH), and it was mainly located in primary spermatocytes (Sc1), secondary spermatocytes (Sc2) and spermatozoa (St). Spats1 expression in embryos was not significantly changed after 17α-methyltestosterone (MT)and 17β-estradiol (E2) treatment. In adults, MT significantly induced Spats1 expression in male P. sinensis. However, the expression of Spats1 in testes was not responsive to E2 treatment. In addition, the expression of Spats1 in females was not affected by either MT or E2 treatment. These results imply that Spats1 is a male-specific expressed gene that is mainly regulated by MT and is closely linked to spermatogenesis and release in P. sinensis.
