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.

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.

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 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.

Higher Dietary Arachidonic Acid Levels Improved the Growth Performance, Gonad Development, Nutritional Value, and Antioxidant Enzyme Activities of Adult Sea Urchin(Strongylocentrotus intermedius)

The gonads of sea urchins(Strongylocentrotus intermedius) are characterized by high levels of arachidonic acid(ARA, 20:4 n-6) and eicosapentaenoic acid(EPA, 20:5 n-3). However, to our knowledge, little information is available regarding the physiological response of adult sea urchins to dietary ARA. In the present study, four dietary feeds were formulated with graded ARA(0, 0.5%, 1%, and 2% dry diet). Each diet was randomly allocated to three cages during a 56-day feeding experiment. The results showed that the sea urchin weight gain rate(WGR) and the gonadosomatic index(GI) significantly increased as ARA was equal to or above 1.0% of dry diet(P < 0.05). The activities of superoxide dismutase(SOD), catalase(CAT), and total anti-oxidative capacity(T-AOC) were the highest in the coelomic fluid of sea urchins that were fed diets with 1% ARA. The total essential amino acid(TEAA) and its ratio to total non-essential amino acid(TNEAA) showed a similar tendency to WGR and GI as dietary ARA increased, and the highest TEAA and TEAA/TNEAA were observed in the gonads of sea urchins that were fed diets with 1% ARA. Levels of ARA and ARA/EPA of the gonads increased while n-3/n-6 polyunsaturated fatty acid(PUFA) decreased with the increase of dietary ARA(P < 0.05). EPA in the gonads of experimental animals fed with formulated feeds showed no significant differences(P> 0.05), but was significantly lower than those fed with kelp(P < 0.05). These results suggested that relatively higher levels of ARA(1% dry diet) significantly promoted growth, gonad development, activities of antioxidant enzymes, as well as nutritional values(TEAA, TEAA/TNEAA, and PUFA) of adult S. intermedius.

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.

Localization of nitric oxide synthase in the developing gonads of amphioxus Branchiostoma belcheri tsingtauense

The localization of nitric oxide synthases （NOS） is dealed with in the developing gonads of amphioxus Branchiostoma belcheri tsingtauense. It was found by NADPH-diaphorase staining that （1） NOS activity was present in the nuclear membranes of germinal vesicles during the entire period ofoocyte development; （2） NOS was localized in both the nuclear membranes and the perinuclear region of cytoplasm in the vitellogenetic oocytes; （3） NOS was relocated in the cortical layer in the mature egg; （4） NOS activity was present in spermatocytes, but not in the spermatogonia in the middle of October; （5） NOS was detected in both spermatozoa and spermatids as well as spermatocytes during the breeding season. This is the first report on the distribution pattern of NOS in the developing gonads in protochordates. These results suggest a role for NOS in the functioning of the nuclear membranes and yolk synthesis during oogenesis and in cell division and differentiation during spermatogenesis.

Anti-diabetic and anti-hyperlipidemic effects of sea cucumber(Cucumaria frondosa) gonad hydrolysates in type Ⅱ diabetic rats

The present study investigated the anti-diabetic activity and potential mechanisms of sea cucumber gonad hydrolysates(SCGH)in a rat model of type II diabetes induced by streptozotocin(STZ)combined with high-fat diet(HFD).Results showed that SCGH significantly reduced water intake,fasting blood glucose level and glycated hemoglobin level.Moreover,the oral glucose tolerance,insulin resistance and plasma lipid level in diabetic rats were also alleviated.Furthermore,histological analysis showed that SCGH effectively protected the tissue structure of liver.In addition,mechanism studies showed that SCGH improved glucose metabolism via activating the IRS/Akt signaling pathway,and promoted lipid metabolism via activating the AMPK signaling pathway.In summary,these findings suggested that SCGH have potential anti-diabetic effects by improving insulin resistance and lipid metabolism disorders.

WJSC 6^(th) Anniversary Special Issues(2):Mesenchymal stem cells Differentiation of mesenchymal stem cells into gonad and adrenal steroidogenic cells

Hormone replacement therapy is necessary for patients with adrenal and gonadal failure.Steroid hormone treatment is also employed in aging people for sex hormone deficiency.These patients undergo such therapies,which have associated risks,for their entire life.Stem cells represent an innovative tool for tissue regeneration and the possibility of solving these problems.Among various stem cell types,mesenchymal stem cells have the potential to differentiate into steroidogenic cells both in vivo and in vitro.In particular,they can effectively be differentiated into steroidogenic cells by expressing nuclear receptor 5A subfamily proteins(steroidogenic factor-1 and liver receptor homolog-1)with the aid of cAMP.This approach will provide a source of cells for future regenerative medicine for the treatment of diseases caused by steroidogenesis deficiencies.It can also represent a useful tool for studying the molecular mechanisms of steroidogenesis and its related diseases.

Seasonal variations in gonad morphology and hypothalamic GnRH-I and GnIH in Eurasian Tree Sparrow,a multi-brooded passerine

The hypothalamic-pituitary-gonadal(HPG)axis ubiquitously regulates seasonal reproduction,following the rhythmicity of a suite of environmental cues.Birds display prominent seasonal variations in gonad size regulated by two hypothalamic peptides,gonadotropin-releasing hormone-1(GnRH-I)and gonadotropin-inhibiting hormone(GnIH).How multi-brooded avian species adjust GnRH-I and GnIH to regulate gonadal morphology seasonally remains largely unknown.Here,we studied the variations in the hypothalamic immunoreactivity(ir)of GnRH-I and GnIH,gonadal proliferation,and apoptosis in a typical multi-brooded species,the Eurasian Tree Sparrow(Passer montanus),across the pre-breeding(PB),first breeding(FB),second breeding(SB),pre-basic molt(PM),and wintering stages(WS).Our results showed that both sexes had higher preoptic area(POA)-GnRH-I-ir but lower paraventricular nucleus(PVN)-GnIH-ir neurons during the breeding stages(FB and SB)relative to other life-history stages,with no significant differences between two broods.The testes and ovaries were significantly larger during the breeding stages.Testicular volume increased during the two broods due to anincreased diameter of seminiferous tubules.Furthermore,there were more testicular apoptotic cells in PB and WS stages than in breeding stages,and in PB stage than in PM stage.Males had higher POA-GnRH-I expression than females during the breeding stages,but both sexes had comparable PVN-GnIH expression throughout the annual cycle.Both sexes of the sparrows may undergo a similar pattern of life-history stage-dependent variation in the hypothalamic GnRH-I,GnIH,and gonadal morphology,except that during breeding stages,males may display higher expression of POA-GnRH-I relative to females.The higher expression of POA-GnRH-I-ir in breeding male sparrows may be critical for male-dependent parental care.

Sex-Based Variation of Gene Expression in the Gonads and Fins of Russian Sturgeon (<i>Acipenser gueldenstaedtii</i>)

Russian sturgeon (Acipenser gueldenstaedtii) is a primitive freshwater fish and a source of black caviar. The genes involved in sexual determination and differentiation are still unknown and there are no molecular markers for sex identification in this species. Studying the variation of the sex-based differences in genomic sequences and in gene expression in the sturgeon may lead to markers of sex in early stages of development and advances in aquaculture, as well as provide novel insights about the evolution of reproduction, sex determination, and sexual differentiation mechanisms in vertebrates. Previous studies by our and other groups have identified differentially expressed genes in the gonads of adult female and male sturgeon. The current study aimed to test whether these genes were also differentially expressed in non-gonadal tissue, namely fins. We measured by qRT-PCR the mRNA levels of 29 known and novel sex-related genes in the gonads and fins of males (4 years old) and females (7 years old;sexual maturation is earlier in males than in females). Six genes (ATP6, IGFRM, LIA1A, S1A, NPL1A, GAPDH and SOX9) showed higher expression in female fins. However, only ATP6 mRNA levels differed in fins of males and females of the same age (4 years old). These findings underscore the impracticality of sex identification based on gene expression in non-gonadal tissue and the need for genetic sex markers in the Russian sturgeon.

Expression of hepatitis B virus surface antigens induces defective gonad phenotypes in Caenorhabditis elegans

AIM To test whether a simple animal, Caenorhabditis elegans(C. elegans), can be used as an alternative model to study the interaction between hepatitis B virus antigens(HBs Ag) and host factors. METHODS Three plasmids that were able to express the large, middle and small forms of HBs Ags(LHBs Ag, MHBs Ag, and SHBs Ag, respectively) driven by a ubiquitous promoter(fib-1) and three that were able to express SHBs Ag driven by different tissue-specific promoters were constructed and microinjected into worms. The brood size, egglaying rate, and gonad development of transgenic worms were analyzed using microscopy. Levels of m RNA related to endoplasmic reticulum stress, enpl-1, hsp-4, pdi-3 and xbp-1, were determined using reverse transcription polymerase reaction(RT-PCRs) in three lines of transgenic worms and dithiothreitol(DTT)-treated wild-type worms. RESULTS Severe defects in egg-laying, decreases in brood size, and gonad retardation were observed in transgenic worms expressing SHBs Ag whereas moderate defects were observed in transgenic worms expressing LHBs Ag and MHBs Ag. RT-PCR analysis revealed that enpl-1, hsp-4 and pdi-3 transcripts were significantly elevated in worms expressing LHBs Ag and MHBs Ag and in wild-type worms pretreated with DTT. By contrast, only pdi-3 was increased in worms expressing SHBs Ag. To further determine which tissue expressing SHBs Ag could induce gonad retardation, we substituted the fib-1 promoter with three tissue-specific promoters(myo-2 for the pharynx, est-1 for the intestines and mec-7 for the neurons) and generated corresponding transgenic animals. Moderate defective phenotypes were observed in worms expressing SHBs Ag in the pharynx and intestines but not in worms expressing SHBs Ag in the neurons, suggesting that the secreted SHBs Ag may trigger a cross-talk signal between the digestive track and the gonad resulting in defective phenotypes. CONCLUSION Ectopic expression of three forms of HBs Ag that causes recognizable phenotypes in transgenic worms suggests that C. elegans can be used as an alternative model for studying virus-host interactions because the resulting phenotype is easily detected through microscopy.

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.

Expression Characteristics of Odf3 in Gonad Are Different Between Chlamys farreri and Vertebrates

Outer dense fibers are specific subcellular components of mammal sperm flagella, apart from the axoneme and its associated proteins. As one of the outer dense fibers components, ODF3 is essential for the formation of mammal sperm flagella. In the present study, we isolated the Odf3 gene, 1033 bp in length, from Zhikong scallop Chlamys farreri, which was named as Cf-Odf3. It was predicted that Cf-Odf3 encodes a protein of 240 amino acids which contains 5 discontinuous Pro-Gly-Pro repeats. The Cf-Odf3 transcripts were detected in both testis and ovary of C. farreri at various stages through q RT-PCR. The Cf-ODF3 protein synthesized by prokaryotes was purified using Ni^(2+)-NTA affinity chromatography and used to produce its polyclonal antibody. The antibody product was effective and specific as was examined by Western blotting. The Cf-Odf3 transcript and Cf-ODF3 protein were located in intragonadal somatic cells(ISCs), spermatogonia and spermatocytes of C. farreri testes, while no transcription of Cf-Odf3 and synthesis of Cf-ODF3 were detected in spermatids and spermatozoa. The location of Odf3 expression is different from that of vertebrates where ODF3 is synthesized specifically in spermatids and sperm flagella. Moreover, the Cf-Odf3 expression was also detected in ISCs, oogonia and oocytes of the scallop ovaries. Our finding revealed a different characteristic of Odf3 expression between scallop and vertebrates, which implied that Cf-Odf3 played a role in the gametogenesis of C. farreri.

METHOD FOR OBSERVATION OF DEEMBEDDED SECTIONS OF FISH GONAD BY SCANNING ELECTRON MICROSCOPY

This article reports a method for examining the intracellular structure of fish gonads using a scanning electron microscope(SEM). The specimen preparation procedure is similar to that for transmission electron microscopy wherein samples cut into semi thin sections are fixed and embedded in plastic. The embedment matrix was removed by solvents. Risen free specimens could be observed by SEM. The morphology of matured sperms in the gonad was very clear, and the oocyte internal structures appeared in three dimensional images. Spheroidal nucleoli and yolk vesicles and several bundles of filaments adhered on the nucleoli could be viewed by SEM for the first time.

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.

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.

Identification of global alternative splicing and sex-specific splicing via comparative transcriptome analysis of gonads of Chinese tongue sole(Cynoglossus semilaevis)

Chinese tongue sole(Cynoglossus semilaevis)is an economically important marine fish species with a ZZ/ZW sex determination mechanism,which can be influenced by temperature.Alternative splicing(AS)is an important mechanism regulating the expression of genes related to sex determination and gonadal differentiation,but has rarely been reported in fish.In this study,to explore the molecular regulatory mechanisms of sex determination and gonadal differentiation,we combined isoform and RNA sequencing(Iso-Seq and RNA-Seq)to perform transcriptome profiling of male and female gonads in C.semilaevis.In total,81883 and 32341 full-length transcripts were obtained in males and females,respectively.A total of 8279 AS genes were identified,including 2639 genes showing differential AS(DAS)between males and females.Many intersecting DAS genes and differentially expressed genes(DEGs)were enriched in the meiotic cell cycle pathway,and genes related to gonadal differentiation,such as esrrb and wt1a,were found to have sex-specific isoforms.Thus,this study revealed AS events in the gonadal transcriptomes of male and female C.semilaevis,described the characteristics of active transcription in the testes,and identified candidate genes for studying the regulatory mechanisms of AS during gonadal differentiation.

Histological Observation of Gonads of Male Elk (Elaphurus davidianus)

Elk (Elaphurus davidianus) is a rare species native to China, and much less efforts have been dedicated to gonadal histology of male elk.The microscopic morphology of gonadal tissue of elk was observed by paraffin method. The results showed that the tunica albuginea testis was com-posed of collagenous fibers and elastic fibers, with the thickness of about 1 054 μm, and a large number of blood vessels were distributed in tunica albuginea;the seminiferous tubule was about 203 μm in diameter, with uneven thickness of tubule wall, which was consisted of sustentacular cells and spermatogenic cells;there were interstitial cells and blood vessels between seminiferous tubules. The epididymis contained more than 20 effer-ent ducts and a long curly epididymal duct;the capsule thickness at epididymis head was about 53 μm, and that at epididymis body and tail was400-1 000 μm;the epithelium of efferent duct was pseudostratified columnar epithelial cells;the wall epithelium of epididymal duct was composed of tall columnar cells at proximal lumina and stroma cells at distal lumina;the epithelial basement membrane of efferent duct and epididymal duct contained circular smooth muscle. The wall of spermatic duct was divided into mucosae, tunica muscularis and tunica adventitia. The tunica muscu-laris was well developed, which was smooth muscle fiber.