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.

Engineering of ovarian tissue for ovarian dysfunctions:A review

This review explores tissue engineering as a potential solution for reproductive health issues in women caused by genetic or acquired diseases,such as premature ovarian failure or oophorectomy.The loss of ovarian function can lead to infertility,osteoporosis,and cardiovascular disease.Hormone replacement therapy is a common treatment,but it has limitations and risks.The review focuses on two main approaches in tissue engineering:scaffold-based(3D printing,electrospinning,decellularization)and scaffold-free(stem cell transplantation,organoid cultivation).Both approaches show promise in preclinical studies for creating functional ovarian tissue.Challenges include vascularization,innervation,long-term function,and safety.Despite these challenges,tissue engineering offers a potential avenue for restoring fertility and hormone balance in women with ovarian dysfunction.

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.

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.

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.

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.

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.

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 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.

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.

Sex-biased autophagy as a potential mechanism mediating sex differences in ischemic stroke outcome

Stroke is the second leading cause of death and a major cause of disability worldwide,and biological sex is an important determining factor in stroke incidence and pathology.From childhood through adulthood,men have a higher incidence of stroke compared with women.Abundant research has confirmed the beneficial effects of estrogen in experimental ischemic stroke but genetic factors such as the X-chromosome complement can also play an important role in determining sex differences in stroke.Autophagy is a self-degrading cellular process orchestrated by multiple core proteins,which leads to the engulfment of cytoplasmic material and degradation of cargo after autophagy vesicles fuse with lysosomes or endosomes.The levels and the activity of components of these signaling pathways and of autophagy-related proteins can be altered during ischemic insults.Ischemic stroke activates autophagy,however,whether inhibiting autophagy after stroke is beneficial in the brain is still under a debate.Autophagy is a potential mechanism that may contribute to differences in stroke progression between the sexes.Furthermore,the effects of manipulating autophagy may also differ between the sexes.Mechanisms that regulate autophagy in a sex-dependent manner in ischemic stroke remain unexplored.In this review,we summarize clinical and pre-clinical evidence for sex differences in stroke.We briefly introduce the autophagy process and summarize the effects of gonadal hormones in autophagy in the brain and discuss X-linked genes that could potentially regulate brain autophagy.Finally,we review pre-clinical studies that address the mechanisms that could mediate sex differences in brain autophagy after stroke.

Gonad traits and age structure of the sea urchin Heliocidaris crassispina in central range:a comparison with rangeextenders

The sea urchin Heliocidaris crassispina has northern expanded in geographic range to Toga Bay,Oga Peninsula in the Sea of Japan due to the global warming.To demonstrate the gonad traits and age structure of H.crassispina in the central range and compare the difference with those in northern extended range,we collected 100 H.crassispina individuals from a barren off Shitsumi,Fukui(central range),in August 2018.Their growth,gonad traits(size,development and color),and sex ratio were investigated and compared with those in Toga Bay,in August 2014(extended range).The successive 2012–2016 year classes of H.crassispina indicated successive juvenile recruitment in the central range,as found in the extended range.The individuals at three years old in central range were obviously larger than those in the extended range,showing large test diameter and body weight,possibly due to the higher temperature in central range than that in northern extended range.In addition,a balanced sex ratio in the central range was found,in contrast to the female-skewed sex ratio found in the extended range.Gonadal development was sex-and site-dependent.The testicular development in the central range was not only delayed in comparison to ovarian development,but also less delayed than those in the extended range,which possibly attribute to the higher water temperature in the central range.A significantly lower male gonad index and L*(lightness)value was recorded in central ranges compared to that in extended range,possibly due to the releasement of sperm of H.crassispina in central ranges.

Poultry genetic heritage cryopreservation and reconstruction:advancement and future challenges

Poultry genetics resources,including commercial selected lines,indigenous breeds,and experimental lines,are now being irreversibly lost at an alarming rate due to multiple reasons,which further threats the future livelihood and academic purpose.Collections of germplasm may reduce the risk of catastrophic loss of genetic diversity by guaranteeing that a pool of genetic variability is available to ensure the reintroduction and replenishment of the genetic stocks.The setting up of biobanks for poultry is challenging because the high sensitiveness of spermatozoa to freezing–thawing process,inability to cryopreserve the egg or embryo,coupled with the females being heterogametic sex.The progress in cryobiology and biotechnologies have made possible the extension of the range of germplasm for poultry species available in cryobanks,including semen,primordial germ cells,somatic cells and gonads.In this review,we introduce the state-of-the-art technologies for avian genetic resource conservation and breed reconstruction,and discuss the potential challenges for future study and further extending of these technologies to ongoing and future conservation efforts.

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.

男性性别决定：深入了解分子机制

Disorders of sex development often arise from anomalies in the molecular or cellular networks that guide the differentiation of the embryonic gonad into either a testis or an ovary, two functionally distinct organs. The activation of the Y-linked gene Sry （sex- determining region Y） and its downstream target Sox9 （Sry box-containing gene 9） triggers testis differentiation by stimulating the differentiation of Sertoli cells, which then direct testis morphogenesis. Once engaged, a genetic pathway promotes the testis development while actively suppressing genes involved in ovarian development. This review focuses on the events of testis determination and the struggle to maintain male fate in the face of antagonistic pressure from the underlying female programme.