Sex Reversal Research Articles

Macrophage activation drives ovarian failure and masculinization in zebrafish.

BMP15 is a conserved regulator of ovarian development and maintenance in vertebrates. In humans, premature ovarian insufficiency is caused by autoimmunity and genetic factors, including mutation of BMP15. The cellular mechanisms underlying ovarian failure caused by BMP15 mutation and immune contributions are not understood. Using zebrafish, we established a causal link between macrophage activation and ovarian failure, which, in zebrafish, causes sex reversal. We define a germline-soma signaling axis that activates macrophages and drives ovarian failure and female-to-male sex reversal. Germline loss of zebrafish Bmp15 impairs oogenesis and initiates this cascade. Single-cell RNA sequencing and genetic analyses implicate ovarian somatic cells that express conserved macrophage-activating ligands as mediators of ovarian failure and sex reversal. Genetic ablation of macrophages or elimination of Csf1Rb ligands, Il34 or Csf1a, delays or blocks premature oocyte loss and sex reversal. The axis identified here provides insight into the cells and pathways governing oocyte and ovary maintenance and potential therapeutic targets to preserve female fertility.

Spatial variation of the broodstock availability of Acanthopagrus berda (Forsskal 1775) with emphasis on seasonality, lunar periodicity, and sex ratio for facilitating efficient breeding program

Successful breeding program for hermaphrodites needs planned broodstock collection. The presented work aims to study the dynamics of broodstock availability of Acanthopagrus berda along selected stretches of Indian coast to facilitate scientific broodstock management. Broodstock availability was recorded for three consecutive years from two geographic locations, Karwar [offshore (beach area) using shore seine and inshore (estuarine area) using hook and lines, cast net and gillnet] and Calicut (inshore using hook and lines, cast net and gillnet). In both regions, most of the stocks were collected between the post-monsoon and pre-monsoon seasons (August to March). At Calicut, a considerable quantity of broodstock was available during all lunar phases except the new moon phase, while at Karwar, it was available except for the first and third quarter and full moon with assured availability during the waxing and waning phases. At Karwar, in the shore seine, the broodstock was available only from August to November, but in cast net, the broodstock was available from August to February. The brooders collected from the Calicut coast were significantly (Welch's statistics p < 0.05, Hedge's g = 0.79) larger (25.476 ± 0.32 cm) than those obtained from the Karwar (21.64 ± 0.17 cm). The highest mean length was observed in the case of fish collected by hook and line, cast net and gill net at Calicut (25.47 ± 0.31), followed by cast net (21.79 ± 0.52) and shore seine (21.60 ± 0.17) at Karwar. The estimated length of sex reversal (L50) was 25.50 cm total length (TL) with a considerable overlap in sex category in the size range of 20 − 30 cm. Welch ANOVA (One-Way) for difference in mean length affirmed no significant difference, and hence the probability of getting a higher number of a particular sex during a given lunar phase could not be assumed.

Temperature- and genotype-dependent stress response and activation of the hypothalamus-pituitary-interrenal axis during temperature-induced sex reversal in pejerrey Odontesthes bonariensis, a species with genotypic and environmental sex determination

In the pejerrey Odontesthes bonariensis (Atheriniformes, Atherinopsidae), exposure to high and low temperatures during the critical period of sex determination (CPSD) induce testicular and ovarian differentiation, respectively, regardless of the presence or not of the sex determining gene amhy, which is crucial for testis formation only at intermediate, sexually neutral temperatures. In this study we explored the existence of genotype-specific signaling of Crh (Corticotropin Releasing Hormone) family genes and their associated carrier protein, receptors, and other stress-related genes in response to temperature during the CPSD and the potential involvement of the central nervous system via the hypothalamus-pituitary-interrenal (HPI) axis in the sex determination of this species. The Crh family genes crhb, uts1, ucn3, the receptor crhr1 and the stress-related genes gr1, gr2, nr3c2 were transiently upregulated in the heads of pejerrey larvae during the CPSD by high temperature alone or in combination with other factors. Only crhr2 transcript abundance was not influenced by temperature but independently by time and genotype. In most cases, mRNA abundance was higher in the XX heads compared to that of XY individuals. The mRNAs of some of these genes were localized in the hypothalamus of pejerrey larvae during the CPSD. XX larvae also showed higher whole-body cortisol titers than the XY, downregulation of cyp19a1a and upregulation of the testis-related genes amhy/amha in trunks (gonads) and were 100% masculinized at the high temperature. In contrast, at the low temperature, crhbp and avt were upregulated in the heads, particularly the former in XY larvae. cyp19a1a and amhy/amha were up- and downregulated, respectively, in the gonads, and fish were 100% feminized. Signaling via the HPI axis was observed simultaneously with the first molecular signs of ongoing sex determination/differentiation in the gonads. Overall, the results strongly suggest a temperature-dependent, genotype-specific regulatory action of the brain involving the Crh family of stress-related genes on the process of environmental sex determination of pejerrey.

Alternative splicing of histone demethylase Kdm6bb mediates temperature-induced sex reversal in the Nile tilapia

Sex determination in many fish species is remarkably plastic and temperature sensitive. Nile tilapia display a genetic sex-determination system (XX/XY). However, high-temperature treatment during critical thermosensitive periods can induce XX females into XXm pseudo-males, and this phenomenon is termed temperature-induced sex reversal (TISR). To investigate the molecular mechanism of TISR in Nile tilapia, we performed Iso-seq analysis and found a dramatic effect of high temperature on gene alternative splicing (AS). Kdm6bb histone demethylase showed a novel AS at intron 5 that generates Kdm6bb_tv1 transcripts without intron 5 and Kdm6bb_tv2 with intron 5. Kdm6bb_tv1 encodes a full-length protein while Kdm6bb_tv2 encodes a truncated protein. Expression analysis revealed that intron 5 splicing of Kdm6bb is male and gonad biased at larval stage, and only gonad biased at adult stage. High-temperature treatment induced intron 5 splicing in the gonads of XX and XY fish, resulting in increased Kdm6bb_tv1 expression. To directly test the role of Kdm6bb_tv1 in Nile tilapia TISR, we knocked out expression of Kdm6bb_tv1. However, Kdm6bb_tv1-/- homozygous mutants showed embryonic lethality. Overexpression of Kdm6bb_tv1, but not Kdm6bb_tv2, induced sex reversal of XX females into pseudo-males. Overexpression of Kdm6bb_tv1, as with high-temperature treatment, modified the promotor region of Gsdf and Dmrt1 by demethylating the trimethylated lysine 27 of histone 3 (H3K27me3), thereby increasing expression. Collectively, these studies demonstrate that AS of Kdm6bb intron 5 increases the expression of Kdm6bb_tv1, which acts as a direct link between high temperature and activation of Gsdf and Dmrt1 expression, leading to male sex determination.

CRISPR/Cas9-mediated gene mutation of EcIAG leads to sex reversal in the male ridgetail white prawn Exopalaemon carinicauda.

In the culture of crustaceans, most species show sexual dimorphism. Monosex culture is an effective approach to achieve high yield and economic value, especially for decapods of high value. Previous studies have developed some sex control strategies such as manual segregation, manipulation of male androgenic gland and knockdown of the male sexual differentiation switch gene encoding insulin-like androgenic gland hormone (IAG) in decapods. However, these methods could not generate hereditable changes. Genetic manipulation to achieve sex reversal individuals is absent up to now. In the present study, the gene encoding IAG (EcIAG) was identified in the ridgetail white prawn Exopalaemon carinicauda. Sequence analysis showed that EcIAG encoded conserved amino acid structure like IAGs in other decapod species. CRISPR/Cas9-mediated genome editing technology was used to knock out EcIAG. Two sgRNAs targeting the second exon of EcIAG were designed and microinjected into the prawn zygotes or the embryos at the first cleavage with commercial Cas9 protein. EcIAG in three genetic males was knocked out in both chromosome sets, which successfully generated sex reversal and phenotypic female characters. The results suggest that CRISPR/Cas9-mediated genome editing technology is an effective way to develop sex manipulation technology and contribute to monosex aquaculture in crustaceans.

Endocrine regulation of reproductive biology in echinoderms: An evolutionary perspective from closest marine invertebrate relatives to chordates

Echinoderms are a phylum of invertebrate deuterostomes, which contain echinoids, asteroids, holothuroids, crinoids, and ophiuroids. Echinoderms have special evolutionary position and unique characteristics, including pentamerous radial body structure, elaborate calcareous endoskeletons, and versatile water vascular system. Echinoderms exhibit extraordinarily diverse reproductive modes: asexual reproduction, sexual reproduction, sexual reversal, etc. Endocrine regulation plays important well-known roles in sex differentiation, gonadal development and maturation, gametogenesis, and reproductive behavior in vertebrates. However, the entire picture of reproductive endocrinology in echinoderms as an evolutionary model of the closest marine invertebrate relatives to chordates has not been revealed. Here, we reviewed previous and recent research progress on reproductive endocrinology in echinoderms, mainly including two sections: Sex steroids in echinoderms and neuropeptide regulation in echinoderm reproduction. This review introduces a variety of endocrine regulatory mechanisms in reproductive biology of echinoderms. It discusses the vertebrate-like sex steroids, putative steroidogenic pathway and metabolism, and reproduction-related neuropeptides. The review will provide a deeper understanding about endocrine regulatory mechanisms of gonadal development in lower deuterostomes and the application of endocrine control in economic echinoderm species in aquaculture.

Sex steroid hormones and steroid secretory cells in the gonads of cyclostomes and fish

. The purpose of this work is to generalize data on the content of sex steroid hormones in cyclostomes and fish in connection with the different state of the gonads, as well as on the localization and ultrastructural organization of steroid secretory cells (SCs). Method used: generalization of literature data. Novelty: for the first time, data on the development of SC in the gonads of juvenile fish are presented on the scale of a literature review; changes in their localization and functional activity are shown. Results: Most of the data on the steroidogenic function of the gonads in fish were obtained in connection with the study of sex differentiation (inversion) and sexual maturation (spawning). The issues of the synthesis of sex steroid hormones and their molecular structure, blood levels in various states of the gonads, as well as the localization and ultrastructural organization of SCs are considered. It has been noted that SCs appear in the gonads long before sex differentiation and can be present among stromal, granulosa, and theca cells. In cyclostomes and fish — juvenile protogynous hermaphrodites in the gonads of genetic males, SCs do not appear in the membranes around the oocytes of the previtellogenesis period, which determines their deficiency of estrogen hormones and, as a result, sex inversion. With induced sex reversal, the development of the steroidogenic function of the gonads proceeds in the opposite direction to its natural development: in males, oocytes appear and steroid activity shifts from the stroma of the gonads into granulosa and theca; in females, on the contrary, secretory activity in the follicular membranes decreases and increases in the stroma of the gonads. Practical significance: the results of the analysis will give insight into the nature of sex steroid hormones, their dynamics in the blood, and their role in the implementation of various processes of gonadogenesis and gametogenesis before their practical use in fish farms.

The -KTS splice variant of WT1 is essential for ovarian determination in mice.

Sex determination in mammals depends on the differentiation of the supporting lineage of the gonads into Sertoli or pregranulosa cells that govern testis and ovary development, respectively. Although the Y-linked testis-determining gene Sry has been identified, the ovarian-determining factor remains unknown. In this study, we identified -KTS, a major, alternatively spliced isoform of the Wilms tumor suppressor WT1, as a key determinant of female sex determination. Loss of -KTS variants blocked gonadal differentiation in mice, whereas increased expression, as found in Frasier syndrome, induced precocious differentiation of ovaries independently of their genetic sex. In XY embryos, this antagonized Sry expression, resulting in male-to-female sex reversal. Our results identify -KTS as an ovarian-determining factor and demonstrate that its time of activation is critical in gonadal sex differentiation.

Regulatory mechanism of LncRNAs in gonadal differentiation of hermaphroditic fish, Monopterusalbus

BackgroundMonopterusalbus is a hermaphroditic fish with sex reversal from ovaries to testes via the ovotestes in the process of gonadal development, but the molecular mechanism of the sex reversal was unknown.MethodsWe produced transcriptomes containing mRNAs and lncRNAs in the crucial stages of the gonad, including the ovary, ovotestis and testis. The expression of the crucial lncRNAs and their target genes was detected using qRT‒PCR and in situ hybridization. The methylation level and activity of the lncRNA promoter were analysed by applying bisulfite sequencing PCR and dual-luciferase reporter assays, respectively.ResultsThis effort revealed that gonadal development was a dynamic expression change. Regulatory networks of lncRNAs and their target genes were constructed through integrated analysis of lncRNA and mRNA data. The expression and DNA methylation of the lncRNAs MSTRG.38036 and MSTRG.12998 and their target genes Psmβ8 and Ptk2β were detected in developing gonads and sex reversal gonads. The results showed that lncRNAs and their target genes exhibited consistent expression profiles and that the DNA methylation levels were negatively regulated lncRNA expression. Furthermore, we found that Ptk2β probably regulates cyp19a1 expression via the Ptk2β/EGFR/STAT3 pathway to reprogram sex differentiation.ConclusionsThis study provides novel insight from lncRNA to explore the potential molecular mechanism by which DNA methylation regulates lncRNA expression to facilitate target gene transcription to reprogram sex differentiation in M.albus, which will also enrich the sex differentiation mechanism of teleosts.

Production of sterile mono-sex triploid yellow drum (Nibea albiflora): genotypic females and sex-reversed phenotypic males with emphasis on utilization as surrogate broodstock.

Production of sterile mono-sex fish is of great significance for sustainable aquaculture as well as germ cell transplantation. In this study, we aimed to produce mono-sex triploid yellow drum, including genotypic females (XXX female) and sex-reversed phenotypic males (XXX male). Firstly, the mono-female triploids were produced through cold-shock treatment on eggs fertilized with sperm from neo-males. Then, the mono-male triploids were produced by the sex reversal of mono-female triploids with oral administration of letrozole (LZ). We comparatively investigated the growth and gonadal development in the mono-sex triploids. The results showed that the triploids displayed similar growth performance to their diploids throughout their first year, but had impaired gonadosomatic index and gametogenesis. No mature gametes were produced in the triploids during their first spawning season. Meanwhile, we analyzed the process of gametogenesis in the both sex of triploids. Ultrastructure of gametogenesis showed that the germ cells arrested at abnormal metaphase 1 in females, while males had irregular meiotic divisions, variable-sized spermatid and degenerated cells. The expression levels of meiosis-related genes (i.e., sycp3 and rec8) confirmed the abnormal meiosis in the triploids. Furthermore, the gonadal development was also determined by the expression patterns of vasa, dmrt1 and cyp19a1a. Abnormal expression of vasa mRNA and protein were detected in triploids. High cyp19a1a expression levels suggested the sex steroid hormones production might be at least partially functional in triploid females. In addition, high dmrt1 expression levels confirmed the masculinization and testicular development of sex-reversed triploid males by LZ. Our findings provide an efficient protocol to produce sterile mono-sex triploid yellow drum and provide new insights into the mechanism of gonadal sterility of triploid fish.

Tilapia, a good model for studying reproductive endocrinology

The Nile tilapia (Oreochromis niloticus), with a system of XX/XY sex determination, is a worldwide farmed fish with a shorter sexual maturation time than that of most cultured fish. Tilapia show a spawning cycle of approximately 14 days and can be artificially propagated in the laboratory all year round to obtain genetically all female (XX) and all male (XY) fry. Its genome sequence has been opened, and a perfect gene editing platform has been established. With a moderate body size, it is convenient for taking enough blood to measure hormone level. In recent years, using tilapia as animal model, we have confirmed that estrogen is crucial for female development because 1) mutation of star2, cyp17a1 or cyp19a1a (encoding aromatase, the key enzyme for estrogen synthesis) results in sex reversal (SR) due to estrogen deficiency in XX tilapia, while mutation of star1, cyp11a1, cyp17a2, cyp19a1b or cyp11c1 affects fertility due to abnormal androgen, cortisol and DHP levels in XY tilapia; 2) when the estrogen receptors (esr2a/esr2b) are mutated, the sex is reversed from female to male, while when the androgen receptors are mutated, the sex cannot be reversed; 3) the differentiated ovary can be transdifferentiated into functional testis by inhibition of estrogen synthesis, and the differentiated testis can be transdifferentiated into ovary by simultaneous addition of exogenous estrogen and androgen synthase inhibitor; 4) loss of male pathway genes amhy, dmrt1, gsdf causes SR with upregulation of cyp19a1a in XY tilapia. Disruption of estrogen synthesis rescues the male to female SR of amhy and gsdf but not dmrt1 mutants; 5) mutation of female pathway genes foxl2 and sf-1 causes SR with downregulation of cyp19a1a in XX tilapia; 6) the germ cell SR of foxl3 mutants fails to be rescued by estrogen treatment, indicating that estrogen determines female germ cell fate through foxl3. This review also summarized the effects of deficiency of other steroid hormones, such as androgen, DHP and cortisol, on fish reproduction. Overall, these studies demonstrate that tilapia is an excellent animal model for studying reproductive endocrinology of fish.

Histomorphic analysis and expression of mRNA and miRNA in embryonic gonadal differentiation in Chinese soft-shelled turtle (Pelodiscus sinensis)

The Chinese soft-shelled turtle (Pelodiscus sinensis) is extensively cultured in Asia for its nutritional and medical value. Gonadal differentiation is fantastic in turtles, whereas morphologic, mRNA, and miRNA expressions were insufficient in the turtle. In this study, ovaries and testes histomorphology analysis of 14–23 stage embryos were performed, and mRNA and miRNA expression profiles were analyzed. Histomorphology analysis revealed that gonads were undifferentiated at embryonic stage 14. Ovarian morphological differentiation became evident from stage 15, which was characterized by the development of the cortical region and degeneration of the medullary region. Concurrently, testicular morphological differentiation was apparent from stage 15, marked by the development of the medullary region and degeneration of the cortical region. qRT-PCR results showed that Cyp19a1 and Foxl2 exhibited female-specific expression at stage 15 and the expression increased throughout most of the embryonic development. Dmrt1, Amh, and Sox9 displayed male-specific expression at stage 15 and tended to increase substantially at later developmental stages. The expression of miR-8356 and miR-3299 in ZZ gonads were significantly higher than that in ZW gonads at stage 15, 17 and 19, and they had the highest expression at stage 15. While the expression of miR-8085 and miR-7982 had the highest expression at stage 19. Furthermore, chromatin remodeler genes showed differential expression in female and male P. sinensis gonads. These results of master sex-differentiation genes and morphological characteristics would provide a reference for the research of sex differentiation and sex reversal in turtles. Additionally, the expression of chromatin remodeler genes indicated they might be involved in gonadal differentiation of P. sinensis.

Effects of dietary arachidonic acid on growth, gonadal development, and tissue fatty acid composition in the hermaphroditic swamp eel (Monopterus albus)

10-week dietary arachidonic acid (ARA) feeding trials were conducted in 14-month-old swamp eel (Monopterus albus) to evaluate the effects on growth performance, gonadal development, tissue fatty acid profile, and gene expression related to lipid metabolism. The results showed that swamp eels fed diets supplemented with 1.4% or 2.8% ARA had a significantly higher weight gain rate (WGR) and specific growth rate (SGR) than those fed the control diet. Dietary supplementation with ARA improved oocyte development and maturation. Higher gonadosomatic index (GSI) was detected in 1.4% and 2.8% ARA groups. ARA supplementation delayed sexual reversal in females. The ARA content in the muscle, liver and gonads increased with increasing dietary ARA concentration; however, ARA supplementation significantly reduced the transcriptional expression of lipid metabolism-related genes in the gonads, such as fad2, pla2g4a, and elovl5. Serum alanine aminotransferase (ALT) levels in the ARA 1.4% group were significantly lower than those in the control and ARA 2.8% groups. Total cholesterol (T-CHO) and triglyceride (TG) levels were significantly lower in the 1.4% and 2.8% ARA groups than in the control group. Catalase (CAT) and high-density lipoprotein cholesterol (HDL-C) increased with dietary ARA concentrations. This study found that dietary ARA is important for improving growth, Ovarian development, and antioxidant capacity in swamp eels.

Bidirectional sex-change behavior and physiological aspects in the Gorgeous goby Lythrypnus pulchellus (Gobiidae).

The Gorgeous goby Lythrypnus pulchellus shows extreme sexual plasticity with the bidirectional sex-change ability socially controlled in adults. Therefore, this study describes how the hierarchical status affects hormone synthesis through newborn hormone waste products in water and tests the influence of body size and social dominance establishment in sex reversal duration and direction. The associated changes in behavior and hormone levels are described under laboratory conditions in male-male and female-female pairs of similar and different body sizes, recording the changes until spawning. The status establishment occurred in a relatively shorter time period in male and female pairs of different sizes (1-3 days) compared to those of similar size (3-5 days), but the earlier one did not significantly affect the overall time of sex change (verified by pair spawning). The changes in gonads, hormones, and papilla occurred in sex-changer individuals, but the first one was observed in behavior. Courtship started at 3-5 days in male pairs and from 2 h to 1 day in female pairs of both groups of different and similar sizes. Hormones did not gradually move in the new sexual phenotype direction during the sex-change time course. Nonetheless, estradiol regulated sex change and 11-ketotestosterone enabled bidirectional sex change and was modulated by agonistic interactions. Cortisol is associated with status and gonadal sex change. In general, similar mechanisms underlie sex change in both directions with a temporal change sequence in phases. These results shed new light on sex-change mechanisms. Further studies should be performed to determine whether these localized changes exist in the steroid hormone synthesis along the brain-pituitary gonad axis during social and bidirectional sex changes in L. pulchellus.

Growth, sex reversal pattern, and reproductive characteristics of Barramundi (Lates calcarifer) broodstock candidates reared in floating cages

Sex reversal of male to female is a characteristic of barramundi (Lates calcarifer), which is affected by several factors, thereby changing the broodstock population. A study was conducted in floating cages in Langkawi, Malaysia, to determine the weight point at the onset of the sex reversal phenomena. A total of 75 female and 55 male adult individuals (3–4 weeks of age) were sampled from the fish cultured in cages to ascertain their sex at different weights. The water temperature and salinity values were 29.82 °C and 33.12 ppt, respectively. The specimens were classified into twelve bodyweight classes (2.00–8.00 ± 0.5 kg intervals). Female specimen body weight distribution was highest in the 6.01–6.50 kg class (22.6%), followed by the 5.51–6.00 kg and 4.51–5.00 class (13.3%), while male specimen body weight distribution was highest in the 4.51–5.00 kg class (32.1%), followed by the 4.01–4.50 kg class (30.3%). Length-to-weight relationships for females and males of Asian Seabass indicated positive allometric growth. The correlation between body weight and GSI, using Pearson’s correlation, for both sexes, for the male and female barramundi, there was a weak correlation between body weight and GSI, which was 37 and 30%, respectively. Based on the present study’s findings, it can be concluded that sex reversal from male to female in Barramundi largely occurred at 4.57 kg body weight and 66.8 cm total length.

The Effects of Mucuna pruriens and Basella alba Extracts on Sex Reversal in Rainbow Trout (Oncorhynchus mykiss)

The use of synthetic steroids for sex reversal in various fish is widespread, but due to the potential dangers of these types of compounds, the use of phytochemicals has been explored as a suitable natural alternative. In this experiment, the effects of extracts of Mucuna pruriens seeds and Basella alba leaves on sex reversal in Oncorhynchus mykiss were investigated. Fifteen-days-old rainbow trout larvae (mean weight: 0.123 ± 0.001 g) were fed experimental diets containing 17 α-methyltestosterone (MT) (60, 90 mg/kg diet), ethanol extract of B. alba leaves (1.0, 2.0 g/kg diet), and methanol extract of M. pruriens seeds (0.1, 0.2 g/kg diet) for 90 days. After 90 days, the experimental diets were replaced with the basal diet until 10 months of age. The control group was fed the basal diet. At the end of the experiment, growth factors, survival (%), intersex (%), and masculinization (%) were measured. The feed conversion ratio and specific growth rate in all experimental groups were not significantly different from those in the control group ( P &gt; 0.05 ). The highest significant percentage of intersex (6.03 ± 3.53%) and the lowest percentage of survival (72.33 ± 2.51%) were observed in the treatment with 90 mg MT/kg compared with the other treatments ( P &gt; 0.05 ). The percentage of intersexuality was significantly lower in the groups fed with plant extract than in the groups fed with MT ( P &lt; 0.05 ). The percentage of intersexuality was significantly lower in the groups fed with plant extract than in the groups fed with MT. The highest percentage of masculinization was observed in the groups fed with M. pruriens extract (0.2 g/kg) (61.75 ± 17.16%) and MT (90 mg/kg) (68.62 ± 1.82%). Moreover, masculinization was significantly higher in all experimental groups than in the control group ( P &gt; 0.05 ). There were significantly more males (%) in all experimental groups than in the control group. In conclusion, extracts of M. pruriens and B. alba can be used as an environmentally friendly and safe alternative to MT to induce masculinization of rainbow trout.

Gametogenesis of pacific salmon. 1. Pink salmon &lt;i&gt;Oncorhynchus gorbuscha&lt;/i&gt;

Cited information on gametogenesis of pink salmon Oncorhynchus gorbuscha Walbaum is summarized. Periods of indifferent development and sex differentiation are considered, development of the testes and ovaries is traced, and the fertility formation is analyzed. Regularities of sex reversion, unique for salmonids, and evolutionarily acquired specific features of oogenesis associated with a short life cycle are noted.

The fundamentals of fibroblast growth factor 9

Fibroblast growth factor 9 (FGF9) was first identified during a screen for factors acting on cells of the central nervous system (CNS). Research over the subsequent two decades has revealed this protein to be a critically important and elegantly regulated growth factor. A hallmark control feature is reciprocal compartmentalization, particularly during development, with epithelium as a dominant source and mesenchyme a prime target. This mesenchyme selectivity is accomplished by the high affinity of FGF9 to the IIIc isoforms of FGFR1, 2, and 3. FGF9 is expressed widely in the embryo, including the developing heart and lungs, and more selectively in the adult, including the CNS and kidneys. Global Fgf9-null mice die shortly after birth due to respiratory failure from hypoplastic lungs. As well, their hearts are dilated and poorly vascularized, the skeleton is small, the intestine is shortened, and male-to-female sex reversal can be found. Conditional Fgf9-null mice have revealed CNS phenotypes, including ataxia and epilepsy. In humans, FGF9 variants have been found to underlie multiple synostoses syndrome 3, a syndrome characterized by multiple joint fusions. Aberrant FGF9 signaling has also been implicated in differences of sex development and cancer, whereas vascular stabilizing effects of FGF9 could benefit chronic diseases. This primer reviews the attributes of this vital growth factor.

Insulin-like Androgenic Gland Hormone Induced Sex Reversal and Molecular Pathways in Macrobrachium nipponense: Insights into Reproduction, Growth, and Sex Differentiation.

This study investigated the potential to use double-stranded RNA insulin-like androgenic gland hormone (dsIAG) to induce sex reversal in Macrobrachium nipponense and identified the molecular mechanisms underlying crustacean reproduction and sex differentiation. The study aimed to determine whether dsIAG could induce sex reversal in PL30-male M. nipponense during a critical period. The sex-related genes were selected by performing the gonadal transcriptome analysis of normal male (dsM), normal female (dsFM), neo-female sex-reversed individuals (dsRM), and unreversed males (dsNRM). After six injections, the experiment finally resulted in a 20% production of dsRM. Histologically, dsRM ovaries developed slower than dsFM, but dsNRM spermathecae developed normally. A total of 1718, 1069, and 255 differentially expressed genes were identified through transcriptome sequencing of the gonads in three comparison groups, revealing crucial genes related to reproduction and sex differentiation, such as GnRHR, VGR, SG, and LWS. Principal Component Analysis (PCA) also distinguished dsM and dsRM very well. In addition, this study predicted that the eyestalks and the "phototransduction-fly" photoperiodic pathways of M. nipponense could play an important role in sex reversal. The enrichment of related pathways and growth traits in dsNRM were combined to establish that IAG played a significant role in reproduction, growth regulation, and metabolism. Finally, complete sex reversal may depend on specific stimuli at critical periods. Overall, this study provides valuable findings for the IAG regulation of sex differentiation, reproduction, and growth of M. nipponense in establishing a monoculture.

Rescue of bmp15 deficiency in zebrafish by mutation of inha reveals mechanisms of BMP15 regulation of folliculogenesis.

As an oocyte-specific growth factor, bone morphogenetic protein 15 (BMP15) plays a critical role in controlling folliculogenesis. However, the mechanism of BMP15 action remains elusive. Using zebrafish as the model, we created a bmp15 mutant using CRISPR/Cas9 and demonstrated that bmp15 deficiency caused a significant delay in follicle activation and puberty onset followed by a complete arrest of follicle development at previtellogenic (PV) stage without yolk accumulation. The mutant females eventually underwent female-to-male sex reversal to become functional males, which was accompanied by a series of changes in secondary sexual characteristics. Interestingly, the blockade of folliculogenesis and sex reversal in bmp15 mutant could be partially rescued by the loss of inhibin (inha-/-). The follicles of double mutant (bmp15-/-;inha-/-) could progress to mid-vitellogenic (MV) stage with yolk accumulation and the fish maintained their femaleness without sex reversal. Transcriptome analysis revealed up-regulation of pathways related to TGF-β signaling and endocytosis in the double mutant follicles. Interestingly, the expression of inhibin/activin βAa subunit (inhbaa) increased significantly in the double mutant ovary. Further knockout of inhbaa in the triple mutant (bmp15-/-;inha-/-;inhbaa-/-) resulted in the loss of yolk granules again. The serum levels of estradiol (E2) and vitellogenin (Vtg) both decreased significantly in bmp15 single mutant females (bmp15-/-), returned to normal in the double mutant (bmp15-/-;inha-/-), but reduced again significantly in the triple mutant (bmp15-/-;inha-/-;inhbaa-/-). E2 treatment could rescue the arrested follicles in bmp15-/-, and fadrozole (a nonsteroidal aromatase inhibitor) treatment blocked yolk accumulation in bmp15-/-;inha-/- fish. The loss of inhbaa also caused a reduction of Vtg receptor-like molecules (e.g., lrp1ab and lrp2a). In summary, the present study provided comprehensive genetic evidence that Bmp15 acts together with the activin-inhibin system in the follicle to control E2 production from the follicle, Vtg biosynthesis in the liver and its uptake by the developing oocytes.