Single-cell RNA sequencing reveals gonadal dynamic changes during sex differentiation in hermaphroditic protogynous orange-spotted grouper(Epinephelus coioides)

DEAR EDITOR,Sex differentiation is a complex process that requires many factors to regulate gonadal proliferation, differentiation,development, and organization. In teleosts, the molecular mechanisms of sex differentiation are diverse and unclear.

A FAMILIAL 46 XY GONADAL DYSGENESIS AND HIGH INCIDENCE OF EMBRYONIC GONADAL TUMORS

The Swyer’s syndrome, 46XY gonadal dysgenesis ( 46XYGD) or XY female, belongs to the category of sexual abnomality. The syndrome is characterized by a female phenotype with streak gonads, 46XY karyotype and complicated by frequent develop of gonadal tumors. A congragated familial 46XYGD was analyzed. We have examined the whole family from many aspects and have followed the affected members more than 10 years. In present case , all eight siblings phenotypically were girls ; the two eldest members died between 1970 1972 from embryonic gonadal tumors. Five of remaining 6 girls were affected by the syndrome of 46 XYGD and two have got the same kind of tumor dysgerminoma. By cytogenetic and cytobiological studies and on the basis of accumulated data the mechanism of gonadal carcinogenesis was analyzed. The origin of dysgerminoma for 46XYGD syndrome was postulated.

Effects of spectral composition,photoperiod and light intensity on the gonadal development of Atlantic salmon Salmo salar in recirculating aquaculture systems(RAS)

Artificial lighting regimes have been successfully used to inhibit sexual maturity of Atlantic salmon in confinement.However,when these operations are applied in commercial recirculating aquaculture systems(RAS) using standard lighting technology,sexual maturation is not suppressed.In this study,an L_9(3~3) orthogonal design was used to determine the effects of three factors(spectral composition,photoperiod,and light intensity) on the gonadal development of Atlantic salmon in RAS.We demonstrated that the photoperiod at the tested levels had a much greater effect on the gonadosomatic index and female Fulton condition factor than spectral composition and light intensity.The photoperiod had a significant effect on the secretion of sex steroids and melatonin(P<0.05),and a short photoperiod delayed sex steroid and melatonin level increases.The three test factors had no significant effects on the survival rate,specific growth rate,relative weight gain,and male Fulton condition factor(P>0.05).The optimum lighting levels in female and male Atlantic salmon were LD 8:16,455 nm(or 625 nm),8.60 W/m^2;and LD 8:16,8.60 W/m^2,455 nm respectively.These conditions not only delayed gonadal development,but also had no negative effects on Atlantic salmon growth in RAS.These results demonstrate that a combination of spectral composition,photoperiod and light intensity is effective at delaying the gonadal development of both male and female salmon in RAS.

Recovery of gonadal development in tiger puffer Takifugu rubripes after exposure to 17β-estradiol during early life stages

The aim of the present study was to investigate the long-term effects of 17l）-estradiol （E2） exposure on gonadal development in the tiger puffer （Taktfugu rubripes）, which has a genetic sex determination system of male homogametic XY-XX. Tiger puffer larvae were exposed to 1, 10 and 100 μg/L E2 from 15 to 100 days post-hatch （dph） and then maintained in clean seawater until 400 dph. Changes in sex ratio, gonadal structure and gonadosomatic index （GSI） were monitored at 100, 160, 270 and 400 dph. Sex-associated single nucleotide polymorphism （SNP） markers were used to analyze the genetic sex of samples, except those at 100 dph. Exposure had a positive effect on the conversion of genetically male gonads into phenotypically female gonads at 100 dph. However, gonads from 60% of genetic XY males in the 1-μg/L E2 group and 100% in the 10-μg/L E2 group developed intersexual gonads at 160 dph; gonads of all genetic XY males in the two treatment groups reverted to testis by 270 dph. While 38%, 57% and 44% of gonads of XY fish in the 100-gg/L E2 group reverted to intersexual gonads at 160, 270 and 400 dph, respectively, none reverted to testis after E2 treatment. In addition, E2 exposure inhibited gonadal growth of both genetic sexes, as indicated by the clear dose-dependent decrease in GSI at 270 and 400 dph. The results showed that exposure to E2 during the early life stages of tiger puffer disrupted gonadal development, but that fish recovered after migration to clean seawater. The study suggests the potential use of tiger puffer as a valuable indicator species to evaluate the effects of environmental estrogens on marine fish, thereby protecting valuable fishery resources.

Steroid sex hormone dynamics during estradiol-17β induced gonadal differentiation in Paralichthys olivaceus(Teleostei)

Steroid sex hormones, such as estradiol-17β （E2） and testosterone （T）, are important regulators of sex change in fish. In this study, we examined the effects of E2 treatment on the dynamics of E2 and T during gonadal differentiation in the olive flounder Paralichtbys olivaceus using histology and radioimmunoassay （RIA）. Flounder larvae were divided into five groups （G0~G4）, and fed with 0 （control）, 0.2, 2, 20 and 100 mg E2/kg feed from 35 to 110 day post hatching （dph）. Fish growth in the G1 and G2 groups was not significantly different from that of the control group （P〉0.05）, while fish in the G3 and G4 groups were less active and showed growth depression and high mortality. The gonads of fish in the G3 and G4 groups were smaller and surrounded by hyperplastic connective tissue. The frequency of females in the G0-G4 groups was 54.5%, 75.0%, 100%, 100% and 93.3%, respectively. The RIA analyses of E2 and T showed that T levels decreased during gonadal differentiation, and increased slightly at the onset of ovarian differentiation, while E2 levels increased gradually and peaked at the onset of ovarian differentiation in the control group. In the E2-treated groups, T levels decreased before the onset of ovarian differentiation. E2 levels were high on the 48 dph, but declined to a lower level on the 54 dph, and then increased gradually during gonadal differentiation. And a sharp increase of E2 levels were observed in all E2-treated groups at the onset of ovarian differentiation. The data suggest that T and E2 play important roles during gonadal differentiation, and an E2 dose of 2 mg/kg feed could induce sex reversal in P olivaceus.

Gonadal dysgenesis in Turner syndrome with Y-chromosome mosaicism:Two case reports

BACKGROUND Turner syndrome(TS)has a variety of different karyotypes,with a wide range of phenotypic features,but the specific karyotype may not always predict the phenotype.TS with Y chromosome mosaicism may have mixed gonadal dysgenesis,and the mosaicism is related to the potential for gonadoblastoma.CASE SUMMARY In this case report,we report two cases of TS with different karyotypes and gonadal dysgenesis.Patient 1 had obvious virilization,and was positive for the SRY gene,but her karyotype in peripheral blood lymphocytes was 45X.Patient 2 had a mosaic karyotype,45X/46X,dic(Y:Y)(p11.3:p11.2),and the proportion of Y-bearing cells was 50%in peripheral blood lymphocytes,but the patient had normal female external genitalia and streaky gonads,with no genital virilism.Different tissues in the same TS individual may exhibit different ratios of mosaicism.The gonadal determination and differentiation of mosaic TS are primarily dependent on the predominant cell line in the gonads.CONCLUSION In TS patients with virilization,it is necessary to test at least two to three tissues to search for cryptic Y material.

Annual change of the gonadal development of the amphioxus in Xiamen

The study was made by observing the annual change of the gonadal development of amphioxus in Xiamen cohtinuously for two years from November, 1986 to November, 1988. We determined the gonadal diameter, gonadosomatic index of each amphioxus, and observed the histological characteristic of the gonadal development of the female and male by cut section, which provided the basis for dividing the stages gonads and the determining level of the gonadal development of amphioxus in each month.

Morphological and histological changes in the brains of turbot(Scophthalmus maximus)with gonadal development

The brain plays a critical role in controlling reproduction through the hypothalamus-pituitary-gonadal(HPG)axis in vertebrates.Turbot(Scophthalmus maximus)has become an economically important marine fish in Europe and North China.Previous research investigating turbot reproduction has focused on the role of the HPG axis in regulating egg and sperm production.However,the morphology and histology of the organs in the HPG axis have not been studied.In this study,we investigated the morphology and histology of brains in female and male turbot at different stages of gonadal development.The results showed that the brains of both female and male turbot were composed of seven parts that are typical of advanced teleosts:the telencephalon,diencephalon,cerebellum,hypothalamus,pituitary gland,myelencephalon,and olfactory bulbs.The telencephalon was well-developed and contained five distinct lobes,with the contiguous diencephalon at the caudal portion.The torus longitudinales and rostral torus semicircularis of the mesencephalon flattened along the dorsal surface,and the rostral corpus cerebellum was located in the dorsal portion.The actual total brain volume in mature males was significantly greater(p<0.05)than that of females with gonadal development.Notably,the pituitary volume in male turbot significantly increased(p<0.05)from immature to mature stage,but this difference did not occur in females.The data together illustrate a distinct sex difference in the turbot brain during gonadal development,providing insight into their HPG axes.

Hypothalamic-Pituitary-Gonadal(HPG)Axis and Transcriptional Regulatory Elements Regulate piwil2 Gene Expression During Gametogenesis and Gonadal Development in Japanese Flounder(Paralichthys olivaceus)

The P-element induced wimpy testis(Piwi)proteins,which are associated with PIWI-interacting RNAs(piRNAs),play important roles in meiosis,germ cell division,and germline maintenance.In this study,we identified and characterized the Paralichthys olivaceus piwil2 gene,a constituent factor of the piRNA pathways involved in the biogenesis of reproductive development.The biological analysis indicated that piwil2,which contains PAZ and PIWI domains,was highly conserved between teleosts and tetrapods.The piwil2 distribution profile in different tissues confirmed a sexually dimorphic expression pattern,with a higher expression level in testis.In situ hybridization demonstrated that piwil2 was expressed in the oogonia and oocytes of the ovaries as well as in the Sertoli cells and spermatocytes of the testes.Gene piwil2 showed a maternally inherited expression pattern during embryonic development,and was highly expressed during the early embryonic development.Different luciferase reporters were constructed to determine the transcriptional regulatory mechanisms of piwil2.The piwil2 core promoter region was located at−360 bp to−60 bp.Furthermore,some representative sex hormones,including human chorionic gonadotropin,17α-methyltestosterone,and estradiol-17βhad distinct regulatory effects on piwil2.In a summery,these results indicate that piwil2,regulated by sex hormones and transcriptional elements,has vital functions in the reproductive cycle and gonadal development.

UBE2I stimulates female gonadal differentiation in chicken(Gallus gallus)embryos

Without known analogous sex-determining factors like SRY(sex determining region Y)in mammals,the chicken(Gallus gallus)sex determination mechanism still remains unclear,which highly restricts the biological research on chicken development and poultry single-sex reproduction.Here we not only characterized a new female-biased gene UBE2I and identified the expression pattern by qRT-PCR,but also described the functional role of UBE2I in the gonadal development of chicken embryos.Results showed that UBE2I exhibited a female-biased expression pattern in the early stage of PGCs(primordial germ cells)in embryonic gonads and robust expression in ovaries of newborn chickens.Most importantly,we successfully developed an effective method to interfere or overexpress UBE2I in chicken embryos through the intravascular injection.The qRT-PCR analysis showed that the sex-related genes(FOXL2,CYP19A1 and HINTW)in females were upregulated(P<0.05)under the overexpression of UBE2I and the sex-related genes(SOX9,DMRT1 and WT1)in females were downregulated(P<0.05)after interfering UBE2I.Furthermore,the change of UBE2I expression was associated with the level of estradiol and its receptors(AR and ESR),which suggests that UBE2I is necessary to initiate the female-specific development in chickens.In conclusion,this work demonstrates that UBE2I is a crucial sex differentiation-related gene in the embryonic development of chickens,which provides insights for further understanding the mechanism of sex determination in chickens.

Mechanism of Gonadal Precocity in Cultured Large Yellow Croaker, Pseudosciaena Crocea : A Study of Microstructure and Submicrostructure of Leydig Cell and Sertoli Cell in Testis

Microstructure and submicrostructure of Leydig cell and Sertoli cell in the testis of gonadal precocity and immaturation in cultured large yellow croaker, Pseudosciaena crocea, are studied using histology and electron microscopic technique. The results indicate that the fine structure of the two kinds of cells in different development stages presents an obvious difference. The smooth endoplasmic reticular and tubular mitochondria of Leydig cell and Sertoli cell are well developed in the testis of gonadal precocity, but poorly developed in the testis of immaturation. We suggest that the reason for gonadal precocity in the large yellow croaker may be related to the earlier development and maturation of Leydig cell and Sertoli cell.

Impairment of the Regulation of Gonadal Function in Channa punctatus by Metacid-50 and Carbaryl under Laboratory and Field Conditions

Regulation of gonadal function by gonadotropic hormone (GtH) and gonadotropin-releasing hormone (GnRH) in Channa punctatus was significantly affected by nonlethal levels of Metacid-50 and Carbaryl. Under laboratory conditions, the time-dependent decrease in serum GtH level was higher in Carbaryl-treated fish than in Metacid-50-treated fish. The situation was reversed in the field, with a higher inhibitory effect of Metacid-50 being recorded. On the other hand, pituitary GtH content and GnRH activity were inhibited to a greater extent by Metacid-50 than by Carbaryl under both field and laboratory conditions. The present findings highlight that even low doses of Metacid-50 and Carbaryl are effective enough to cause reproductive damage, as evidenced by homeostatic unbalance of the reproductive regulatory system. 1990 Academic Press. Inc.

A Case Report: Challenge in Treatment of a Rare Condition of Gonadal Agenesis and Bipolar Disease

Background: We report a rare case of congenital bilateral testicular agenesis who presented with a suicide attempt. We pose the question of testosterone therapy could improve his mental health. Case Report: A 36-year-old man was admitted to a psychiatric ward after the suicide attempt. An endocrinology consult is requested to address if hormone replacement therapy could improve his depression and future suicide attempts. His past medical history is significant for gonadal agenesis, testicular implants placement at age 16, bipolar type 1 diagnosed 2 years ago before the presentation. The patient received testosterone replacement treatment at age 13 - 15 years. Since that time, he has not been placed on any testosterone therapy. Physical exam revealed minimal axillary and pubic hair, testicular implants with a microphallus. Laboratory values reflect hypergonadotropic hypogonadism consistent with testicular agenesis. He was suicidal and testosterone treatment was deferred. Discussion: It is a rare case of congenital testicular agenesis with bipolar disorder which was resistant to therapy. The patient had profound hypogonadism, which needed testosterone replacement therapy. However, the therapy was deferred because of an acute psychotic condition. This case also highlights that there is no strong evidence of the effect of hormonal therapy on mental health in this group.

Effect of Mercuric Chloride and Cadmium Chloride on Gonadal Function and Its Regulation in SexuallyMature Common Carp Cyprinus carpio

Gonadal function in fish, CJprinus carpio was significantly affected by sublethal doses of mercuric chloride (HgCl2 ) and cadmium chloride (CdCl2 ) in chronic (45 days ) exposure.Parameters investigated were nonesterified (NE) and esterified (E) cholestcrol of ovary,liver and serum and ovarian 3β-Hydroxysteroid and 17β-Hydroxysteroid dehyrogenaseenzyme activity and serum and pituitary gonadotropin (GtH) levels. Both the pollutantswere able to reduce the hypothalamic extract (HE) or gonadotropin releasing hormone(GnRH) induced pituitary GtH release in vitro. Short term (96h) exposure of the fish tothe pollutants had no significant effect on the gonadal function. ln addition to thedeleterious effect of pollutants on the gonadal steroidogenesis and pituitary gonadotropin release, using [4-14C] cholesterol as a tracer it was found that for 45 days expeure, HgCl2'had an adverse effect on the transport of cholesterol from circulation to ovary.

Gonadal Morphology, Histology and Spermatogenesis of Striped Grunt Fish, Pomadasys Stridens (Forsskal, 1775) (Family: Pomadasyidae)

The present study was conducted on some aspects of histology and morphology of gonadal development and spermatogenesis of the striped piggy fish,Pomadasys stridens from the Arabian Sea near Karachi coast.A preliminary investigative study was carried on genetic-variability,morpho-histological characters to observe the differences between both gonads of both the sexes along with the size of the associated fat bodies.Both the gonads were investigated and classified in seven different stages of maturity.Testes were found asymmetrical in measurement with the right testis larger than the left one.It was found that meiotic activity and spermatid development showed the opposite relationship.The left testis showed a relatively greater activity than the right one.

Effect of Varicocelectomy on Gonadal Function among Patients Reporting with Sexual Dysfunction in Ghana

Background: Long-standing varicocele is often associated with testicular hypoxia and that might worsen Leydig cell function, a significant risk factor for hypogonadism. This may affect both the secretory and endocrine functions of the testis. This study aims to determine the effect of microsurgical sub-inguinal varicocelectomy on gonadal function among men reporting sexual dysfunction in Ghana. Methods: This was an intervention study conducted at the Tamale Teaching Hospital from September 2017 to August 2021. A total of 103 participants were randomized into two groups;the surgery group (n = 52) and the observed group (n = 51). Venous blood samples were collected at baseline, varicocelectomy was performed for the surgery group, and no intervention was given to the other. Blood samples were subsequently collected at 12-, 24-, 36-, and 48-month intervals for assay of serum total testosterone, FSH, and LH. The data were analyzed in GraphPad Prism (v8.0) at an alpha value of 0.05. Results: All the participants had varicocele and were aged between 55.0 to 69.0 years old. At the baseline of the study, all participants presented with sexual dysfunction but a significant improvement (p Conclusions: Microsurgical sub-inguinal varicocelectomy improved gonadal function among varicocele patients reporting sexual dysfunction. It is recommended to use this choice for similar patients;however, these findings should be verified by a multi-institutional study to provide more evidence for this choice.

The m6A methylation regulates gonadal sex differentiation in chicken embryo

Background:As a ubiquitous reversible epigenetic RNA modification,N6-methyladenosine(m6A)plays crucial regulatory roles in multiple biological pathways.However,its functional mechanisms in sex determination and differentiation during gonadal development of chicken embryos are not clear.Therefore,we established a transcriptome-wide m6A map in the female and male chicken left gonads of embryonic day 7(E7)by methylated RNA immunoprecipitation sequencing(MeRIP-seq)to offer insight into the landscape of m6A methylation and investigate the post-transcriptional modification underlying gonadal differentiation.Results:The chicken embryonic gonadal transcriptome was extensively methylated.We found 15,191 and 16,111 m6A peaks in the female and male left gonads,respectively,which were mainly enriched in the coding sequence(CDS)and stop codon.Among these m6A peaks,we identified that 1013 and 751 were hypermethylated in females and males,respectively.These differential peaks covered 281 and 327 genes,such as BMP2,SMAD2,SOX9 and CYP19A1,which were primarily associated with development,morphogenesis and sex differentiation by functional enrichment.Further analysis revealed that the m6A methylation level was positively correlated with gene expression abundance.Furthermore,we found that YTHDC2 could regulate the expression of sex-related genes,especially HEMGN and SOX9,in male mesonephros/gonad mingle cells,which was verified by in vitro experiments,suggesting a regulatory role of m6A methylation in chicken gonad differentiation.Conclusions:This work provided a comprehensive m6A methylation profile of chicken embryonic gonads and revealed YTHDC2 as a key regulator responsible for sex differentiation.Our results contribute to a better understanding of epigenetic factors involved in chicken sex determination and differentiation and to promoting the future development of sex manipulation in poultry industry.

A novel heterozygous SF1/NR5A1 gene variant causes 46,XY DSD-gonadal dysgenesis with hypergonadotropic hypogonadism without adrenal insufficiency

Steroidogenic factor 1(SF1/NR5A1;nuclear receptor subfamily 5 group A member 1)is an essential orphan protein involved in gonadal embryogenesis,sex determination,and reproductive endocrinology.

Mechanisms of Immune Tolerance and Inflammation via Gonadal Steroid Hormones in Preterm Birth

In 2019,preterm births(PTB)accounted for approximately 0.66 million deaths globally.PTB is also associated with a significantly higher risk of mortality and long-term complications for newborns.Long-term studies associated several factors,including disruption of immune tolerance and inflammation,with PTB.However,the pathogenesis of PTB remains unclear.Gonadal steroid hormones are critical for pregnancy maintenance and regulation of immune and inflammatory responses.However,it is not clear how unbalanced gonadal steroid hormones,such as imbalanced estrogen/androgen or estrogen/progesterone contribute to PTB.In this review,we discuss how gonadal steroid hormones mediate dysfunction in immune tolerance and inflammatory responses,which are known to promote the occurrence of PTB,and provide insight into PTB prediction.

The role of sexual related Y gene detection in the diagnosis of patients with gonadal dysgenesis

Objective To clarify the role of sexual related Y (SRY) gene detection in the diagnosis of gonadal dysgenesis.Methods Sixteen cases of gonadal dysgenesis were included in this study: 5 with androgen insensitivity syndrome, 1 with 17-α-hydroxylase deficiency, 4 with true hermaphrodite, 2 with 45,X/46,XY gonadal dysgenesis, 1 with 45,X gonadal dysgenesis, 1 with XY pure gonadal dysgenesis, 1 with testicular regression, and 1 XY female who gave birth to a normal baby. SRY gene was detected by using polymerase chain reaction (PCR) in blood and gonad samples and by direct sequencing of the SRY motif.Results Among the 16 cases, 15 were blood SRY positive, among which 13 (86.7%) showed the presence of testicular tissue, and 2 showed ovaries without testicular tissue. One SRY negative case showed the presence of testicular tissue. In 3 cases, SRY detection in gonadal tissue correlated with pathological findings but not with blood karyotype. The correlation between peripheral blood SRY and the pathology of the gonads was 81.25% and the correlation between the presence of peripheral blood Y chromosome and pathology of the gonads was 68.75%. Sequencing of the SRY motif in an XY female who gave birth to a normal baby showed no mutation.Conclusions SRY detection is more sensitive and specific than blood karyotype in the prediction of the presence of testicular tissue. Peripheral blood karyotype does not necessarily reflect gonadal type. There may be testicular related factors other than the SRY gene.