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The expression of sex determining region of the Y chromosome (Sry) in the fetal gonads is important for male development. In a mouse model of disorders of sex development (C57BL/6 (B6)-XYᴾᴼˢ), the gonadal phenotype and the timing of Sry expression differ due to differences among B6 substrains as the genetic background. Since differences in Sry expression among B6 substrains have been speculated, the present study examined Sry expression in B6J, B6JJmsSlc, and B6NCrl mice. These substrains differed in the number of Sry-expressing cells in the gonads of embryonic mice at each developmental stage, with B6NCrl having more than the other strains. The substrains differed also in the number of Sry-expressing cells between the left and right gonads, with B6J and B6NCrl, but not B6JJmsSlc, showing left gonad-dominant Sry expression. Substrain differences existed also in the distribution of Sry-expressing cells in the medial and lateral directions of gonads. In addition, in the left gonad-dominant Sry-expressing substrains B6J and B6NCrl, the medial and central regions of the left gonad had more Sry-expressing cells than those of the right gonad. Substrains of B6 mice have not always been considered in sex differentiation studies. In the present study, however, we observed substrain differences in the number of Sry-expressing cells, left–right distribution, and medial/lateral distribution during the early stages of gonadal development in B6 mice. Therefore, future studies on sex differentiation in B6 mice should consider substrain differences.

The molecular background of disorders of sex development (DSD) in dogs is poorly understood. Several copies of the SRY genes have been reported in the dog genome. We used droplet digital PCR with the aim of determining variability in SRY copy number and its association with DSD in dogs. Altogether 19 DSD male dogs (XY DSD) of 10 breeds and 87 control dogs of eight breeds were analyzed. Moreover, we performed a comparative analysis of SRY copy number in other canids: wolves (3), red foxes (16), and Chinese raccoon dogs (10). We found that the modal number of SRY copies in dogs, wolves, red foxes, and Chinese raccoon dogs was 3, 3, 1, and 3 respectively. Variability of copy number was only observed in Yorkshire Terriers (two or three copies) and red foxes (one or two copies). An analysis of six DSD Yorkshire Terriers and 38 control males of this breed showed that 50% of the DSD dogs had two copies, while the incidence of this variant was significantly lower in the control dogs (10.5%). Searching for the copy number of the coding and 5'-flanking fragments revealed full concordance with the copy number. These fragments were also sequenced in DSD (19) and control (24) dogs, and no DNA variants were found. We conclude that, in the dog, two or three functional copies of the SRY gene are present, and a smaller number of copies showed an association with the risk of DSD phenotype in Yorkshire Terriers.

BACKGROUND Co-morbidity of SRY gene turner syndrome (TS) with positive SRY gene and non-classical congenital adrenal hyperplasia (NCAH) is extremely rare and has never been reported to date. CASE SUMMARY In this article, we present a 14-year-old girl who was referred to our hospital with short stature (weight of 43 kg and height of 143 cm, < -2 SD) with no secondary sexual characteristics (labia minora dysplasia). Laboratory tests indicated hypergonadotropic hypogonadism with significantly increased androstenedione and 17-hydroxyprogesterone (17-OHP) levels. This was accompanied by the thickening of the extremity of the left adrenal medial limb. The patient’s karyotype was 45,X/46,X, +mar, and cytogenetic analysis using multiplex ligation-dependent probe amplification and high-throughput sequencing indicated that the SRY gene was positive with compound heterozygous mutations in CYP21A2 as the causative gene for congenital adrenal hyperplasia. The sites of the suspected candidate mutations were amplified and verified using Sanger sequencing. The patient was finally diagnosed as having SRY positive TS with NCAH. The patient and her family initially refused medical treatment. At her most recent follow-up visit (age = 15 years old), the patient presented facial hair, height increase to 148 cm, and weight of 52 kg, while androstenedione and 17-OHP levels remained high. The patient was finally willing to take small doses of hydrocortisone (10 mg/d). CONCLUSION In conclusion, upon evaluation of the patient mentioned in the report, we feel that 17-OHP measurement and cytogenetic analysis are necessary for TS patients even in the absence of significant virilization signs. This will play a significant role in guiding diagnosis and treatment.

Lakor goat is an Indonesian indigenous goat breeds that successfully survive and is adaptive in the ecology of Lakor island in Southwest Maluku with high temperatures and curah limited rainfall. Genetic variations in Y-chromosome enable researchers to identify paternal lineages, which are informative for introgressions and migrations. This study aims to characterize genetic variation and phylogenetic relationships of Lakor goats based on male-specific region markers, i.e., sex-determining region-Y (SRY) gene. The genomes of 84 follicle samples of male goats (bucks) collected from Lakor island, were analyzed. The amplified product for SRY was 850 bp in encoding region (exon-1). Genetic variation and paternal phylogenetic relationship were established following sequencing using MEGA version X.0 software. The results of Y-chromosome alignments of the entire sequences identified two polymorphic nucleotides between individual goats within the population. Two polymorphism nucleotides, i.e. site 362 (A-G), and found in LK3, LK7, LK11, and LK13 samples, while site 410 (C-T) was found in LK3, LK7, LK11, LK13, and LK24 samples. The two mutations are categorized as substitution (transition type) and this is in the exon-1 area. This study concluded that genetic variation is very low in the Lakor goat breed and phylogenetic analysis grouping is a single clade or monophyletic group.

BACKGROUND 45,X/46,XY mosaicism is a rare chromosomal abnormality with a wide range of phenotypes in both males and females, from normal individuals with different degrees of genital ambiguity to those who show signs of Turner’s syndrome. More rarely, cases of 45,X/46,XY mosaicism with a normal-appearing male phenotype are not found until a chromosome test is performed to investigate the cause of male infertility. CASE SUMMARY In this study, a 29-year-old male patient with complete azoospermia is reported. Chromosomal analyses of his lymphocytes revealed the karyotype 45,X[93%]/46,X,+mar(Y)[7%]. In addition, Y chromosome-specific markers, such as SRY, ZFY, AZFa, AZFb and AZFc, were not observed in his blood DNA according to multiplex polymerase chain reaction test. A literature review identified several 45,X/46,XY cases with a normal-appearing male phenotype, most of whom were diagnosed during infertility investigation. However, the present case is the first SRY-negative 45,X/46,XY male case diagnosed during a premarital medical examination. CONCLUSION This finding further suggests that sex determination is a complex process regulated by multiple genetic and environmental factors.

Background: The sex determination process requires distinct signaling pathways to generate either testis or ovaries from the same precursor structures, the primordial gonad. Deviations of this signaling mechanism may result in disorders/differences of sex development (DSD). The 46, XX testicular DSD is a rare genetic condition identified by a discrepancy between genetic and phenotypic sex caused sex reversal syndrome. Case Presentation: We describe the case of a 5 years-old 46, XX boy with ambiguous genitalia. On physical examination he had severe hypospadias, bifid scrotum, micropenis and palpable bilateral testes. Cytogenetic analysis of patient reveals a 46, XX karyotype. Hormonal assay showed low level of FSH, LH and Testosterone and there was no evidence of Mullerian structures based on pelvic imaging. The histopathology of gonadal tissue showed a Leydig cell hyperplasia which gives the impression of Sertoli cell nodule. Polymerase chain reaction (PCR) analysis failed to identify the presence of SRY gene, therefore a diagnosis of 46, XX Testicular DSD with SRY-negative was established. Conclusion: This report presents a rare case of SRY-negative 46, XX Testicular DSD in a boy with ambiguous genitalia. A comprehensive management including clinical, cytogenetic and molecular analyses have indicated that undiscovered genetic or environmental factors needs to be elucidated. It is important to carry out further molecular testing to establish precise diagnosis of DSD and to provide appropriate genetic counseling for patients and their family.

Sex determination in mammals is controlled by the dominance of either pro-testis (SRY-SOX9-FGF9) or pro-ovary (RSPO1-WNT4-FOXL2) genetic pathways during early gonad development in XY and XX embryos, respectively. We have previously shown that early, robust expression of mouse Sry is dependent on the nuclear protein GADD45g. In the absence of GADD45g, XY gonadal sex reversal occurs, associated with a major reduction of Sry levels at 11.5 dpc. Here, we probe the relationship between Gadd45g and Sry further, using gain- and loss-of-function genetics. First, we show that transgenic Gadd45g overexpression can elevate Sry expression levels at 11.5 dpc in the B6.YPOS model of sex reversal, resulting in phenotypic rescue. We then show that the zygosity of pro-ovarian Rspo1 is critical for the degree of gonadal sex reversal observed in both B6.YPOS and Gadd45g-deficient XY gonads, in contrast to that of Foxl2. Phenotypic rescue of sex reversal is observed in XY gonads lacking both Gadd45g and Rspo1, but this is not associated with rescue of Sry expression levels at 11.5 dpc. Instead, Sox9 levels are rescued by around 12.5 dpc. We conclude that Gadd45g is absolutely required for timely expression of Sry in XY gonads, independently of RSPO1-mediated WNT signalling, and discuss these data in light of our understanding of antagonistic interactions between the pro-testis and pro-ovary pathways.

Background Permanent progression of paternal age and development of reproductive medicine lead to increase in number of children conceived with assisted reproductive techniques (ART). Although it is uncertain if ARTs have direct influence on offspring health, advanced paternal age, associated comorbidities and reduced fertility possess significant risks of genetic disorders to the offspring. With a broad implementation of a non-invasive prenatal testing (NIPT), more cases of genetic disorders, including sex discordance are revealed. Among biological causes of sex discordance are disorders of sexual development, majority of which are associated with the SRY gene. Case presentation We report a case of a non-invasive prenatal testing and ultrasound sex discordance in a 46,XY karyotype female fetus with an SRY pathogenic variant, who was conceived through an intracytoplasmic sperm injection (ICSI) due to severe oligozoospermia of the father. Advanced mean age of ICSI patients is associated with risk of de novo mutations and monogenic disorders in the offspring. Additionally, ICSI patients have higher risk to harbour infertility-predisposing mutations, including mutations in the SRY gene. These familial and de novo genetic factors predispose ICSI-conceived children to congenital malformations and might negatively affect reproductive health of ICSI-patients’ offspring. Conclusions Oligozoospermic patients planning assisted reproduction are warranted to undergo genetic counselling and testing for possible inherited and mosaic mutations, and risk factors for de novo mutations.

Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms.

Aberrant expression of the SRY-related HMG-box (SOX) genes contributes to tumor development and progression. This research aimed to identify the regulation of the SOX genes in stomach adenocarcinoma (STAD). Expression profiles downloaded from The Cancer Genome Atlas (TCGA) were conducted to analyze the expression and function of the SOX genes. A competing endogenous RNAs (ceRNA) network mediated by the SOX genes was effectively constructed consisting of 64 lncRNAs, 29 miRNAs, and 11 SOX genes based on predicted miRNAs shared by lncRNAs and mRNAs using miRDB, TargetScan, miRTarBase, miRcode, and starBase v2.0. SOX9 was identified as a prognostic signature, which showed the usefulness of diagnosis and prognosis of STAD by the receiver operating characteristic (ROC) and Kaplan-Meier curves. SOX9 was also shown specifically in STAD and identified as highly expressed in the gastrointestinal tract. Gene Ontology (GO) enrichment analysis showed that SOX9 might influence the genes related to the pattern specification process, sodium ion homeostasis, and potassium ion transport, mainly including FEZF1, HOXC13, HOXC10, HOXC9, HOXA11, DPP6, ATP4B, CASQ2, KCNA1, ATP4A, and SFRP1. Furthermore, HOTAIR knockdown, miR-206-mimic transfection, the Cell Count Kit-8 (CCK-8) assay were performed to verify the function of HOTAIR/miR-206/SOX9 axis, which was identified in the ceRNA network analysis. HOTAIR could induce proliferation potentially by competitively binding miR-206/SOX9 axis in STAD. These findings provide new clues with prognostic and therapeutic implications in STAD and suggest that HOTAIR/miR-206/SOX9 might be a potential new strategy for therapeutic targeting of gastric cancer.

Objective: This study aimed to elucidate the effect of miR-140 on the proliferation of porcine fetal fibroblasts (PFFs) and identify the target genes of miR-140 in PFFs.Methods: In this study, bioinformatics software was used to predict and verify target genes of miR-140. Quantitative polymerase chain reaction and western blot were used to detect the relationship between miR-140 and its target genes in PFFs. Dual luciferase reporter gene assays were performed to assess the interactions among miR-140, type 1 insulinlike growth factor receptor (IGF1R), and SRY-box 4 (SOX4). The effect of miR-140 on the proliferation of PFFs was measured by CCK-8 when PFFs were transfected with a miR-140 mimic or inhibitor. The transcription factor SOX4 binding to promoter of IGF1R was detected by chromatin immunoprecipitation assay (ChIP).Results: miR-140 directly targeted IGF1R and inhibited proliferation of PFFs. Meanwhile, miR-140 targeted transcription factor SOX4 that binds to promoter of porcine IGF1R to indirectly inhibit the expression of IGF1R. In addition, miR-140 inhibitor promoted PFFs proliferation, which is abrogated by SOX4 or IGF1R knockdown.Conclusion: miR-140 inhibited PFFs proliferation by directly targeting IGF1R and indirectly inhibiting IGF1R expression via SOX4, which play an important role in the development of porcine fetal.

The ability to identify the sex of embryo and control of sex ratio has a great commercial importance to livestock industry. Prediction of embryonic sex could be useful in the management decisions of sex selection in breeding programs. Several methods have been attempted to determine the sex but the polymerase chain reaction (PCR)-based sexing method is generally favoured, as it is cost effective, simple and reliable. The aim of the present study was to identify sex of sheep embryos produced in vitro through amplification of glyceraldehyde 3-phosphate dehydrogenase (GAPDH), sex-determining region Y (SRY) and amelogenin genes present in genomic DNA (gDNA) of embryos through PCR. To avoid false interpretation of the result by no amplification of SRY in female embryos, a duplex PCR was approached to amplify combinedly SRY and GAPDH genes. Sex-specific blood was used in PCR as positive control. In vitro sheep embryos were produced as per standardized protocol of laboratory. Sexing of sex-specific blood and in vitro produced embryos were approached though PCR to amplify the respective genes using gDNA present in the sample without its traditional isolation. The accuracy of sex prediction for embryos was 100% by this procedure.

International audience; The translocation of SRY onto one of the two X chromosomes results in a 46,XX testicular disorder of sex development; this is supposedly due to non-allelic homologous recombination between the protein kinase X gene (PRKX) and the inverted protein kinase Y pseudogene (PRKY). Although 46,XX SRY-positive men are infertile, the literature data indicate that some of these individuals are of short stature (relative to the general population). We sought to determine whether short stature was linked to additional, more complex chromosomal rearrangements.Twelve laboratories gathered detailed clinical, anthropomorphic, cytogenetic and genetic data (including chromosome microarray (CMA) data) on patients with 46,XX SRY-positive male syndrome.SRY was present (suggesting a der(X)t(X;Y)) in 34 of the 38 cases (89.5%). When considering only the 20 patients with CMA data, we identified several chromosomal rearrangements and breakpoints - especially on the X chromosome. In the five cases for whom the X chromosome breakpoint was located in the pseudoautosomal (PAR) region, there was partial duplication of the derivate X chromosome. In contrast, in the 15 cases for whom the breakpoint was located downstream of the pseudoautosomal region, part of the derivate X chromosome had been deleted (included the arylsulfatase E (ARSE) gene in 11 patients). For patients with vs. without ARSE deletion, the mean height was respectively 167.7 ± 4.5 and 173.1 ± 4.0 cm; this difference was not statistically significant (p = 0.1005).Although 46,XX SRY-positive male syndromes were mainly due to imbalanced crossover between the X and Y chromosome during meiosis, the breakpoints differed markedly from one patient to another (especially on the X chromosome); this suggests the presence of a replication-based mechanism for recombination between non-homologous sequences. In some patients, the translocation of SRY to the X chromosome was associated with ARSE gene deletion, which might have led to short stature. With a view to explaining this disorder of sex development, whole exome sequencing could be suggested for SRY-negative patients. This article is protected by copyright. All rights reserved.

Molecular dynamics (MD) simulations were conducted in order to investigate the effect of the naturally occurring point mutations of the transcription factor (TF) sex-determining region Y (SRY) on the structure and dynamics of the SRY-DNA complex. The normal SRY, along with the two mutants I13T and G40R, comprising point mutations on the SRY chain, which have been clinically identified in patients with sex developmental disorders, were modeled as DNA complexes. Our modeling work aims at elucidating atomic-level structural determinants of the aberrant SRY-DNA complexation by means of μs-long MD. The results suggest that the observed disorders brought about by the G40R-DNA and I13T-DNA may arise predominantly from the destabilization of the complex being in accord with

En la mayoría de los casos, la diferenciación sexual masculina ocurre con la participación del gen SRY. Sin embargo, se pueden presentar otros genotipos excepcionales, como en el caso que se presenta en este reporte.Se trata de un paciente adulto de sexo masculino atendido en el Servicio de Paternidades del Instituto de Genética de la Universidad Nacional de Colombia. Se le hicieron los análisis del gen de la amelogenina y de repeticiones cortas en tándem (Short Tandem Repeat, STR) específicas para el gen SRY con estuches comerciales de identificación humana, así como los de cariotipo convencional e hibridación in situ fluorescente del SRY, y el estudio de microdeleciones del cromosoma Y mediante reacción en cadena de la polimerasa (PCR). Se le hizo la evaluación clínica y se le brindó asesoramiento genético.El paciente no presentaba ambigüedad genital, su cariotipo era 46 XX, y el perfil molecular era negativo para el gen SRY y positivo para el ZFY. Se le diagnosticó un trastorno de diferenciación sexual 46 XX testicular no sindrómico, una rara condición genética. Solo el 20 % de los pacientes con este diagnóstico son negativos para SRY y exhiben perfiles moleculares diversos. La información disponible parece indicar que el ZFY está relacionado con la diferenciación sexual masculina, aún en ausencia del gen SRY.

Introduction: Molecular screening of Y-chromosome AZF microdeletions, along with chromosome analysis, is the first step of the genetic diagnosis routinely performed in infertile men with severe oligozoospermia or non-obstructive azoospermia. These deletions can cover one or more of the three AZF regions, each one with different testicular histological phenotypes ranging from Sertoli cell only (SCO) syndrome (AZFa) to gamete maturation arrest (AZFb) and hypospermatogenesis (AZFc). Complete AZF deletions (AZFabc) are very rare and most likely related to abnormal karyotypes such as 46,XX, 46,X,del(Y)(q11.1) or 46,X,iso(Y)(p10), causing azoospermia with SCO, therefore it is not appropriate to propose testicular sperm extraction due to the impossibility to retrieve testicular spermatozoa for intracytoplasmic sperm injection. Methodology: Since 2000, we analysed 348 samples of azoospermic men, in compliance with EAA/EMQN best practice guidelines for Y-chromosomal microdeletions testing1, and participated annually in the EMQN-EQA programs. Genomic DNA was analysed by multiplex-PCR amplification of STS specific for each AZF region, and SRY and AR (in Xq12) as control markers. In case of AZFabc deletions, an extension PCR-analysis was performed for DYZ3 Y-centromere marker: if present other Yq centromeric proximal border markers were screened; if DYZ3 was absent (most cases of 46,XX males SRY-positive) other Yp specific genes were screened (RPS4Y, ZFY, AMELY, PRKY) in order to characterize the Yp segment. Results/Discussion: We characterized 10 men with AZFabc deletions, 8 of which 46,XX SRYpositive, negative for the centromere and all the AZFabc markers. In these cases, screening for heterochromatin (e.g. sY160) and markers mapping between the centromere and AZFa is inappropriate, as it is highly unlikely that these patients have Yq material between AZFa and the centromere and terminal Yq sequences. Testing for the presence of additional markers besides those used in our methodology do not have any relevance for the diagnosis nor the clinical management of these patients. In such cases, karyotyping evaluation is recommended. 1- Krausz et al 2014, Andrology; 2(1): 5–19 FCT - UIDB/00009/2020 info:eu-repo/semantics/publishedVersion

The sex-determining gene SRY has undergone rapid evolution in rodents. Curiously, a new study by Miyawaki et al. reveals that a recently evolved SRY gene sequence antagonizes SRY protein stability, necessitating splicing of a novel intron. Other data suggest that this troublesome gene region has noncoding RNA functions, possibly related to conflict between sex chromosomes.

Sox9 plays an essential role in mammalian testis formation. It has been reported that gene expression in the testes is regulated by enhancers. Among them, mXYSRa/Enh13—which is located at far upstream of the transcription start site—plays a critical role, wherein its deletion causes complete male-to-female sex reversal in mice. It has been proposed that the binding sites (BSs) of SOX9 and SRY, the latter of which is the sex determining gene on the Y chromosome, are associated with mXYSRa/Enh13. They function as an enhancer, whereby the sequences are evolutionarily conserved and in vivo binding of SOX9 and SRY to mXYSRa/Enh13 has been demonstrated previously. However, their precise in vivo functions have not been examined to date. To this end, this study generated mice with substitutions on the SOX9 and SRY BSs to reveal their in vivo functions. Homozygous mutants of SOX9 and SRY BS were indistinguishable from XY males, whereas double mutants had small testes, suggesting that these functions are redundant and that there is another functional sequence on mXYSRa/Enh13, since mXYSRa/Enh13 deletion mice are XY females. In addition, the majority of hemizygous mice with substitutions in SOX9 BS and SRY BS were female and male, respectively, suggesting that SOX9 BS contributes more to SRY BS for mXYSRa/Enh13 to function. The additive effect of SOX9 and SRY via these BSs was verified using an in vitro assay. In conclusion, SOX9 BS and SRY BS function redundantly in vivo, and at least one more functional sequence should exist in mXYSRa/Enh13.

Testis-determining factor (TDF) is an alternative term for the sex-determining region (SRY) gene found on the Y chromosome. SRY gene mutations can cause a wide range of sex development issues. The single-exon SRY gene is a short and intronless sex-determining gene with a DNA-binding high mobility group (HMG) box that is highly conserved. We performed cytogenetic analysis on one patient with Turner syndrome having a 45,X/46,XY chromosomal composition. Further, we used polymerase chain reaction, single-stranded conformational polymorphism, and automated DNA sequencing to detect a substitution of C→A (missense mutation) at codon 139, which led to an amino acid shift from Proline to Glutamine (P139Q) outside but downstream of the HMG box in the SRY gene. No further alterations in the SRY gene were discovered in the patient. The absence of one X chromosome in 88% of the patient’s cells might have resulted in a wide variety of phenotypic complications.

Little was known about the role of MYCN in drug resistance in acute myeloid leukemia (AML). We investigated impacts of MYCN on sensitization of AML cell to cisplatin, and its interaction with SRY-box transcription factor 4 (SOX4). In vitro, human AML cell lines HL60 and KG-1 were transfected with the overexpression plasmids and specific small interfering RNA of MYCN (siMYCN), or siSOX4, and then treated with 2 μg/mL cisplatin. MTT and flow cytometry assays were performed to estimate the viability and apoptosis of AML cell. The expressions of MYCN and SOX4 in AML patients and the associations between these two were analyzed by bioinformatics analysis, luciferase assay and quantitative reverse transcription polymerase chain reaction (qRT-PCR). In vivo, transfected HL60 cells were injected into BALB/c nude mice. The tumor size and weight were recorded, and the expressions of MYCN and SOX4 were determined with immunohistochemistry. MYCY and SOX4 were highly-expressed in AML, and their expressions were positively correlated. In AML cells, the treatment using cisplatin induced higher MYCN expression and augmented apoptosis, whereas decreasing the viability. Silencing of MYCN enhanced the tumor-suppressive effects of cisplatin on AML by decreasing cell viability and increasing apoptosis. The promoter of SOX4 was targeted by MYCN. Lower cell viability and higher apoptosis rate were seen in AML cells transfected with siSOX4, the effects of which were partially reversed by overexpressed MYCN. Silencing of MYCN retarded the tumor growth and SOX4 expression in tumor in vivo. Collectively, MYCN sensitized AML cells to cisplatin by targeting SOX4.

Oral squamous cell carcinoma (OSCC) is a ubiquitous malignancy and is associated with high mortality. Accumulating evidence indicates that transcription factors play a pivotal role in the progression of OSCC. This study was aimed to investigate the expression of SOX12 in OSCC and its significance. SOX12 expression in OSCC tissues was analyzed through TCGA databases and then tested by Western blot and qRT-PCR analysis. Moreover, SOX12 was silenced by RNA interference in OSCC cells (SCC-25 and SCC-4), and the growth ability of OSCC cells was examined using MTT assay. The level of SOX12 was upregulated in OSCC according to the TCGA results, which was further confirmed in the OSCC cell lines. Patients with high SOX12 expression had shorter overall survival (OS) than those with low SOX12 expression. High expression of SOX12 is positively correlated with T stage of OSCC. In addition, MTT analysis indicated that silencing of SOX12 resulted in reduced OSCC cell proliferation. Taken together, the high expression of SOX12 in OSCC indicates that SOX12 gene may play an essential role in OSCC. Our research indicates that SOX12 expression could be a predictive biomarker and is a potential therapeutic target for OSCC.

An 8-month mixed breed dog (from shelter) was evaluated for its abnormal external genital (enlarged os clitoris protruding externally from the vulva) condition. Dog had phenotypic female appearance. Testicular-like tissue was removed via laparoscopy-assisted gonadectomy. Genotype was determined using blood and gonadal tissue (via histology and immunohistochemistry). Dog had XX chromosomes and was negative for SRY (sex-determining region on the Y chromosome) gene.

Orientadores: Maricilda Palandi de Mello, Celso Eduardo Benedetti, Fernanda Caroline Soardi Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: A expressão do gene SRY (Sex Determining Region in chromosome Y) é responsável por desencadear a determinação testicular durante o desenvolvimento embrionário, a partir das gônadas ainda indiferenciadas. Mutações nesse gene são encontradas em muitos casos de anomalias do desenvolvimento gonadal. O projeto teve por objetivo principal a análise funcional do efeito de uma mutação na região promotora do gene SRY, sendo que essa mutação consiste em uma deleção de 3 pares de base em um dos sítios consenso de ligação do fator de transcrição Sp1 ao promotor do gene. O portador dessa mutação é um indivíduo com disgenesia gonadal pura 46,XY, sendo que outros membros da família apresentavam ambiguidade genital e o pai, também portador da mutação, possuía grave hipospadia ao nascimento. Para tentar esclarecer os efeitos desta mutação nos mecanismos moleculares de regulação da expressão do gene SRY, este trabalho primeiramente analisou a interação da proteína Sp1 com os sítios localizados na região promotora de SRY e o efeito da mutação nesta interação, através de ensaios de EMSA (Electrophoretic Mobility Shift Assay). Concluiu-se que os sítios Sp1A e Sp1B se ligam a duas moléculas de Sp1 e que a mutação no Sp1A praticamente abole esta ligação. Possivelmente a ausência dessa ligação impediu a formação de um complexo de transcrição, causando uma diminuição na expressão do gene SRY e levando à ausência de formação dos testículos e reversão sexual completa na paciente. Complementando, foi analisado o efeito dessa mutação na expressão através de ensaio de expressão com gene repórter, no qual o promotor normal se mostrou em média duas vezes mais eficiente na ativação da expressão da luciferase que o promotor mutante. Entretanto, mais experimentos de transfecção, inclusive com outras linhagens celulares, devem ser realizados para confirmação desse resultado. Além disso, foi analisado o efeito de uma nova mutação (localizada na região codificante do gene SRY) na ligação da proteína SRY com o DNA, através de ensaios de EMSA. Concluiu-se que a mutação E89K, associada com disgenesia gonadal pura 46,XY, reduziu em alto nível a atividade de ligação in vitro da proteína SRY mutante ao DNA, o que representa um forte indício de que atividade reduzida da proteína mutante não foi suficiente para desencadear o processo de determinação testicular. Paralelamente, foi feito o rastreamento de mutações no gene SRY e sua região promotora em novos casos de disgenesia gonadal, não tendo sido encontradas, porém, alterações nos pacientes analisados Abstract: The SRY (Sex Determining Region in chromosome Y) gene expression is responsible for testicular determination during embrionary development. Mutations in SRY are found in many cases of anomalies of gonadal development. This project analyzed the functional effect of a mutation in SRY promoter region; the mutation is a 3-bp deletion in a consensus binding site for Sp1 transcription factor. The patient presented 46,XY pure gonadal dysgenesis, and a family history of relatives with different levels of genital ambiguity. Her father shares the same mutation in the SRY promoter region. In order to investigate the effects of the mutation upon the molecular mechanisms that regulate SRY gene expression, the interaction of the Sp1 transcription factor with normal and mutant binding sites was analyzed by EMSA (Electrophoretic Mobility Shift Assay). Each of Sp1A and Sp1B normal sites binds to a single Sp1 molecule, whereas the 3-bp deletion in Sp1A abolishs the binding to this site. Probably, the lack of Sp1 binding to the Sp1A site prevented the formation of a stable transcription complex, reducing the level of SRY expression and leading to the absence of testicles and complete sex reversal in the patient. Parallely, the effect of the mutation was analyzed by a reporter gene assay, indicating that the normal promoter is almost two times more efficient than the mutant promoter in the activation of luciferase gene expression using HeLa cells. However, further transfection experiments with other cell lineages must be performed to confirm this result. In addition, the effect of a new mutation (E89K, located in the SRY gene coding region) was analyzed by testing the ability of the SRY mutant protein to bind its DNA consensus sequence. EMSA assays revealed that the E89K mutation, which is associated with 46,XY pure gonadal dysgenesis, strongtly reduced the SRY protein binding activity in vitro. This result is a strong evidence that the reduced activity of SRY mutant protein was not sufficient to trigger the testicular determination in the patient, leading to the pure gonadal dysgenesis phenotype. Screening of mutations in the SRY gene coding and promoter regions was also performed in six diferent patients with 46,XY gonadal dysgenesis. However, other mutations have not been identified Mestrado Genética Animal e Evolução Mestre em Genética e Biologia Molecular

Although important factors governing the meiosis have been reported in the embryonic ovary, meiosis in postnatal testis remains poorly understood. Herein, we first report that SRY‐box 30 (Sox30) is an age‐related and essential regulator of meiosis in the postnatal testis. Sox30‐null mice exhibited uniquely impaired testis, presenting the abnormal arrest of germ‐cell differentiation and irregular Leydig cell proliferation. In aged Sox30‐null mice, the observed testicular impairments were more severe. Furthermore, the germ‐cell arrest occurred at the stage of meiotic zygotene spermatocytes, which is strongly associated with critical regulators of meiosis (such as Cyp26b1, Stra8 and Rec8) and sex differentiation (such as Rspo1, Foxl2, Sox9, Wnt4 and Ctnnb1). Mechanistically, Sox30 can activate Stra8 and Rec8, and inhibit Cyp26b1 and Ctnnb1 by direct binding to their promoters. A different Sox30 domain required for regulating the activity of these gene promoters, providing a “fail‐safe” mechanism for Sox30 to facilitate germ‐cell differentiation. Indeed, retinoic acid levels were reduced owing to increased degradation following the elevation of Cyp26b1 in Sox30‐null testes. Re‐expression of Sox30 in Sox30‐null mice successfully restored germ‐cell meiosis, differentiation and Leydig cell proliferation. Moreover, the restoration of actual fertility appeared to improve over time. Consistently, Rec8 and Stra8 were reactivated, and Cyp26b1 and Ctnnb1 were reinhibited in the restored testes. In summary, Sox30 is necessary, sufficient and age‐associated for germ‐cell meiosis and differentiation in testes by direct regulating critical regulators. This study advances our understanding of the regulation of germ‐cell meiosis and differentiation in the postnatal testis., We reported that Sox30‐null mice exhibit uniquely impaired testis with irregular Leydig cell proliferation and abnormal germ‐cell arrest at zygotene, and the observed testicular impairments were more severe as mice aged. Sox30 controls male germ‐cell meiosis by a “fail‐safe” regulation of retinoic acid signalling, and re‐expression of Sox30 in Sox30‐null mice can successfully restore germ‐cell meiosis and differentiation. Moreover, the restoration of actual fertility appeared to improve over time.

Annotation: Swier's syndrome is a genetic disorder, a variant of hypogonadism with a 46 XY karyotype, in which complete or "pure" dysgenesis of the testicles is observed, a female phenotype is formed with a male genotype. It is a congenital disorder associated with the Y chromosome. Swier's syndrome is usually associated with a genetic variant of testicular dysgenesis associated with mutations in the SRY gene (Sex-determining Region Y) located on the Y-chromosome of most mammals, and is involved in the development of the male-type organism (locus Yp11.3). The SRY gene, localized on the short arm of the Y chromosome, is usually expressed by 7-8 weeks of embryogenesis, triggering a signal for the differentiation of Sertoli cells, germinal epithelium of the seminiferous tubules, and Leydig cells. During physiological embryogenesis, AMH (anti-Müllerian hormone) is released in Sertoli cells, leading to regression of the Müllerian ducts, and in Leydig cells stimulated by placental chorionic gonadotropin (CG), and then by their own luteinizing hormone (LH), an increasing amount of testosterone is secreted. The SRY gene encodes the protein TDF (testisdeterminingfactor), a transcription factor that regulates the expression of other genes, which in turn encodes transcription factors that initiate the development of the male reproductive system in the embryonic period. During spermatogenesis during meiosis, the Y chromosome can lose SRY by transferring this gene to the X chromosome. In the absence of full-fledged genetic material (in particular, the expression of the SRY gene), the formation of functionally active testicular tissue is impossible. The lack of secretion of anti-Müllerian hormone leads to the development of Müllerian ducts, from which the uterus, upper vagina and fallopian tubes are formed. Thus, despite the male 46 XY karyotype, the external and internal reproductive organs of the fetus are formed by the female. In this literature review, the defects of the genes responsible for the varieties and clinical manifestations of Swier's syndrome are analyzed.

Background: Disorders of sex development (DSD) are a medical condition that affects the normal process of sexual Various of the genes needed for gonad development have been identified by investigation of patients with disorders sex development (DSD).Phenotypes of patients with 46,XY DSD range from atonalism in female phenotype with complete external genitalia to male phenotype with testicular regression. Individuals with 46,XYagonadiam show a wide range of clinical features and in some cases, there is not a clear diagnosis for this patients. We presented the clinical and molecular study a patient with 46,XY female without gonadal tissue. Case presentation: A 27-year-old female was attended to our center because of primary amenorrhea. Ultrasonography did not show gonadal tissue including Mullerian structures, uterus, and Wolffian structures. Also, the patient had not streak gonad. We performed cytogenetic study and molecular analysis, including automated sequencing of the entire coding region of SRY gene, in the patient with agonadism. Our result showed 46,XY karyotype. Also, we noticed that molecular mutations in SRY are not identified as a cause of DSD female without a gonadal tissue. Laboratory examination showed that this case is a unique patient with 46,XYfemale agonadism that has no association with previously described. Conclusions: The present case was a patient with 46, XYagonadism without hormonal or kidney defect and we did not detect mutation in SRY gene. To our knowledge, this case is a unique patient with 46,XYagonadism that has no association with previously described. So this case would be helpful for clinicians to assess 46,XY female patients without gonadal tissue.

Rationale: Chromosome karyotype analysis and SRY (sex determined region of Y chromosome) gene detection are routines for the diagnosis of growth hormone deficiency (GHD), but further whole exome gene sequencing occasionally leads to subversive results and unexpected conclusions. Patient concerns: We report a single case of a 7-year-old Chinese boy who had stunted growth since he was 1 year old. He was short in height (height Standard Deviation Score (SDS) was less than 2.9), bilateral scrotal dysplasia and delayed bone age. Diagnosis: His growth hormone (GH) stimulation tests showed GHD. His karyotype analysis and polymerase chain reaction (PCR) analyses indicated a 46, XX disorder of sex development (DSD) without the presence of the SRY gene. Nevertheless, considering that female gonad was not observed in the chest and abdominal magnetic resonance imaging, the whole exome gene sequencing was performed. Sequencing data confirmed the presence of SRY gene sequence and two copies of chromosome X. Later, using different primer sequences for PCR, it showed that the SRY gene was positive. The final diagnosis was a rare case of “46, XX (SRY positive) testicular DSD with GHD”. Interventions: The boy's parents agreed to use recombinant human growth hormone (rhGH) for GHD treatment, the starting dose was 0.035 mg / kg / day. But they disagreed with molecular diagnostics and genomic analysis of the Y chromosome. Outcomes: The boy was treated with rhGH for 3 months and his height increased by 2.2 cm. The patient will be followed-up until the end of his puberty. Lessons: In summary, whole exome gene sequencing overturned the preliminary diagnosis results of karyotype analysis and SRY gene detection, and found that there may be a certain correlation between testicular DSD and GHD.

Amaç: Bu çalışmamızda bölgemizde karşılaştığımız cinsiyet anomalili hastalarda bipotansiyel gonad gelişimi sonrası meydana gelen değişimlerde SRY ve SOX9 genlerinin etkisinin araştırılması ve bu genlerdeki mutasyonların testis ve ovaryum oluşumuna olası etkileri incelenerek elde edilecek verilerin ışığında literatüre yeni bilgilerin kazandırılması hedeflenmektedir. Gereç ve Yöntemler: Üniversitemizin Tıbbi Biyoloji- Genetik Laboratuvarlarına Cinsiyet anomalisi şüphesiyle başvuran 0-45 yaş aralığındaki 60 hastadan alınan kandan izole ettiğimiz DNA örnekleri kullanılarak çalışma gerçekleştirildi. Sitogenetik çalışmalar sonucunda 46, XY çıkan hastalara SRY ve SOX9 gen analizi yapıldı. Hasta bireylerin SRY ve SOX9 gen analizleri RT-PCR ile amplifiye edilerek elde edilen veriler bilgisayar sistemi kullanılarak değerlendirildi. Bulgular: Yaptığımız çalışma sonucunda cinsiyet anomalisi tanısı ile gelen 60 hastadan 46,XY olan bireyler sitogenetik analiz sonuçlarına göre tespit edildi ve bu hastalardaki SRY ve SOX9 gen bölgelerinin varlığı ve yokluğuna bakılarak 44 tane 46,XY bireyden 4 tanesinde SRY gen bölgesi bulunmayıp SOX9 gen bölgesi bulunmuştur Sonuç: SOX9 geni otozomal kromozom üzerinde bulunup SRY genine bağlı aktivite göstermektedir. Bunun sonucunda tüm bireylerde SOX9 geni var olup SRY genine bağlı olarak cinsel organ gelişiminde etkilidir. Bu çalışmada SRY ve SOX9 gen bölgelerinin karyotip sonucu ile ilişkili olduğu bununda cinsiyet gelişimi ve cinsel organ gelişimi üzerinde rol oynamış olabileceği düşünülmektedir. Aim: In this study, it is aimed to investigate the effects of SRY and SOX9 genes in the changes that occur after the development of bipotential gonads in patients with gender anomalies in our region, and to provide new information to the literature in the light of the data to be obtained by examining the possible effects of mutations in these genes on the formation of testis and ovary. Metarials and Methods: The study was carried out using DNA samples isolated from the blood taken from 60 patients aged 0-45 years who applied to the Medical Biology-Genetics Laboratories of our university with the suspicion of gender anomaly. As a result of cytogenetic studies, SRY and SOX9 gene analyzes were performed on patients with karyotype 46, XY. The SRY and SOX9 gene analyzes of the patients were amplified by RT-PCR and the data obtained were evaluated using a computer system. Result: As a result of our study, 46,XY individuals out of 60 patients who came with the diagnosis of gender anomaly were determined according to the results of cytogenetic analysis, and the presence and absence of SRY - SOX9 gene regions in these patients were examined and the SRY gene region was not found in 4 of 44 46,XY individuals, but the SOX9 gene region was found. Conclution: The SOX9 gene is located on the autosomal chromosome and shows activity depending on the SRY gene. As a result, all individuals have the SOX9 gene and it is effective in the development of the genitals depending on the SRY gene. As a result of this study, it is thought that the SRY-SOX9 gene regions are associated with the karyotype result, which may play a role in sex development and sexual organ development. Dicle Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü (DÜBAP) tarafından TIP.20.008 numaralı proje ile desteklenmiştir.

The 46, XX male sex reversal syndrome (SRS) is a rare disease with a gender dysplasia phenotype. Scientists concur that SRS is associated with translocation of the sex-determining region Y gene (SRY). We obtained peripheral blood mononuclear cells (PBMCs)from an 18-year-old male with SRS to generate an induced pluripotent stem cell (iPSC) line (SMUSHi001-A). Karyotyping analysis of the patient PBMCs revealed a normal 46, XX karyotype carrying the SRY gene. Pluripotent markers were successfully detected in SMUSHi001-A which can be differentiated into three germ layers in vitro. This cell line will provide a cell model for exploring the pathogenesis and potential therapeutic methods of SRS.

International audience; Objective: SRY-negative 46,XX testicular and ovotesticular disorders/differences of sex development (T/OTDSD) represent a very rare and unique DSD condition where testicular tissue develops in the absence of a Y chromosome. To date, very few studies have described the phenotype, clinical and surgical management and long-term outcomes of these patients. Particularly, early blockade of the gonadotropic axis in patients raised in the female gender to minimize postnatal androgenization has never been reported.Design: Retrospective description of sixteen 46,XX T/OTDSD patients.Results: Sixteen 46,XX SRY-negative T/OTDSD were included. Most (12/16) were diagnosed in the neonatal period. Sex of rearing was male for six patients and female for ten, while the clinical presentation varied, with an external masculinization score from 1 to 10. Five patients raised as girl were successfully treated with GnRH analog to avoid virilization during minipuberty. Ovotestes/testes were found bilaterally for 54% of the patients and unilaterally for the others (with a contralateral ovary). Gonadal surgery preserved appropriate tissue in the majority of cases. Spontaneous puberty occurred in two girls and one boy, while two boys required hormonal induction of puberty. One of the girls conceived spontaneously and had an uneventful pregnancy. DNA analyses (SNP-array, next-generation sequencing and whole-exome sequencing) were performed. A heterozygous frameshit mutation in the NR2F2 gene was identified in one patient.Conclusions: This study presents a population of patients with 46,XX SRY-negative T/OTDSD. Early blockade of gonadotropic axis appears efficient to reduce and avoid further androgenization in patients raised as girls.

Simple Summary Glioblastoma (GBM) is one of the most common and most aggressive brain tumors with higher prevalence among men than women. Loss of chromosome Y (LOY) in the peripheral blood cells has been associated with increased risk of developing cancer. However, there is a lack of data about LOY in GBM tumor tissue and the potential impact on patients prognosis. Through droplet digital PCR (ddPCR) analysis of 10 markers spread throughout chromosome Y in 105 male GBM patients, we were able to identify deletion of SRY gene as a factor strongly correlating with reduced overall survival. This finding was later corroborated by the analysis of GBM gene expression data collected in TCGA, showing correlation between decreased SRY expression and shortened overall survival. Background: Biological causes of sex disparity seen in the prevalence of cancer, including glioblastoma (GBM), remain poorly understood. One of the considered aspects is the involvement of the sex chromosomes, especially loss of chromosome Y (LOY). Methods: Tumors from 105 isocitrate dehydrogenase (IDH) wild type male GBM patients were tested with droplet digital PCR for copy number changes of ten genes on chromosome Y. Decreased gene expression, a proxy of gene loss, was then analyzed in 225 IDH wild type GBM derived from TCGA and overall survival in both cohorts was tested with Kaplan-Meier log-rank analysis and maximally selected rank statistics for cut-off determination. Results: LOY was associated with significantly shorter overall survival (7 vs. 14.6 months, p = 0.0016), and among investigated individual genes survival correlated most prominently with loss of the sex-determining region Y gene (SRY) (10.8 vs. 14.8 months, p = 0.0031). Gene set enrichment analysis revealed that epidermal growth factor receptor, platelet-derived growth factor receptor, and MYC proto-oncogene signaling pathways are associated with low SRY expression. Conclusion: Our data show that deletions and reduced gene expression of chromosome Y genes, especially SRY, are associated with reduced survival of male GBM patients and connected to major susceptibility pathways of gliomagenesis. Funding Agencies|Erik, Karin and Gosta Selanders Foundation at Uppsala University Hospital; Swedish Cancer Foundation; ALF Grants; Region Ostergotland; Medical Research Council of Southeast SwedenUK Research & Innovation (UKRI)Medical Research Council UK (MRC); LiU Cancer at Linkoping University; Lions Cancer Foundation

Significance The present work characterizes the porcine sex-determining region on the Y chromosome (SRY) gene and demonstrates its pivotal role in sex determination. We provide evidence that genetically male pigs with a knockout of the SRY gene undergo sex reversal of the external and internal genitalia. This discovery of SRY as the main switch for sex determination in pigs may provide an alternative for surgical castration in pig production, preventing boar taint. As the pig shares many genetic, physiological, and anatomical similarities with humans, it also provides a suitable large animal model for human sex reversal syndromes, allowing for the development of new interventions for human sex disorders., The sex-determining region on the Y chromosome (SRY) is thought to be the central genetic element of male sex development in mammals. Pathogenic modifications within the SRY gene are associated with a male-to-female sex reversal syndrome in humans and other mammalian species, including rabbits and mice. However, the underlying mechanisms are largely unknown. To understand the biological function of the SRY gene, a site-directed mutational analysis is required to investigate associated phenotypic changes at the molecular, cellular, and morphological level. Here, we successfully generated a knockout of the porcine SRY gene by microinjection of two CRISPR-Cas ribonucleoproteins, targeting the centrally located “high mobility group” (HMG), followed by a frameshift mutation of the downstream SRY sequence. This resulted in the development of genetically male (XY) pigs with complete external and internal female genitalia, which, however, were significantly smaller than in 9-mo-old age-matched control females. Quantitative digital PCR analysis revealed a duplication of the SRY locus in Landrace pigs similar to the known palindromic duplication in Duroc breeds. Our study demonstrates the central role of the HMG domain in the SRY gene in male porcine sex determination. This proof-of-principle study could assist in solving the problem of sex preference in agriculture to improve animal welfare. Moreover, it establishes a large animal model that is more comparable to humans with regard to genetics, physiology, and anatomy, which is pivotal for longitudinal studies to unravel mammalian sex determination and relevant for the development of new interventions for human sex development disorders.

Context The genetic basis of human sex development is slowly being elucidated, and >40 different genetic causes of differences (or disorders) of sex development (DSDs) have now been reported. However, reaching a specific diagnosis using traditional approaches can be difficult, especially in adults where limited biochemical data may be available. Objective We used a targeted next-generation sequencing approach to analyze known and candidate genes for DSDs in individuals with no specific molecular diagnosis. Participants and Design We studied 52 adult 46,XY women attending a single-center adult service, who were part of a larger cohort of 400 individuals. Classic conditions such as17β-hydroxysteroid dehydrogenase deficiency type 3, 5α-reductase deficiency type 2, and androgen insensitivity syndrome were excluded. The study cohort had broad working diagnoses of complete gonadal dysgenesis (CGD) (n = 27) and partially virilized 46,XY DSD (pvDSD) (n = 25), a group that included partial gonadal dysgenesis and those with a broad “partial androgen insensitivity syndrome” label. Targeted sequencing of 180 genes was undertaken. Results Overall, a likely genetic cause was found in 16 of 52 (30.8%) individuals (22.2% CGD, 40.0% pvDSD). Pathogenic variants were found in sex-determining region Y (SRY; n = 3), doublesex and mab-3–related transcription factor 1 (DMRT1; n = 1), NR5A1/steroidogenic factor-1 (SF-1) (n = 1), and desert hedgehog (DHH; n = 1) in the CGD group, and in NR5A1 (n = 5), DHH (n = 1), and DEAH-box helicase 37 (DHX37; n = 4) in the pvDSD group. Conclusions Reaching a specific diagnosis can have clinical implications and provides insight into the role of these proteins in sex development. Next-generation sequencing approaches are invaluable, especially in adult populations or where diagnostic biochemistry is not possible.

Background Approximately 10–15% of 46,XY disorders of sex development (DSDs) have an SRY mutation residing in the high mobility group (HMG) domain. Here, we present a case of 46,XY DSD caused by a novel missense mutation in the HMG region of SRY rapidly progressing to germ cell tumors (GCTs). Case presentation An adolescent female (15 years old) exhibiting primary amenorrhea was later diagnosed as a 46,XY female with bilateral gonadal dysplasia on the basis of peripheral lymphocyte karyotype 46,XY and a novel missense mutation in SRY (c.281 T > G, p.L94R). The novel missense mutation (c.281 T > G, p.L94R) and its adjacent region were conserved. Protein structure analysis showed that the mutant site was located in the middle of the HMG domain, and the mutant protein had a diminished ability to bind to DNA. Imaging examination revealed an adolescent female with a naive uterus. Laparoscopy and initial pathological examination revealed left gonadal dysplasia and right gonadal dysplasia with gonadoblastoma (GB). Right gonadectomy by laparoscopy was performed upon consent from the patient’s parents. Less than 1 year postoperatively, the left gonadal gland deteriorated as observed by the findings of a mass in the left adnexal region by pelvic MRI and serum AFP > 1000 ng/ml by serological tests, and then total hysterectomy and adnexal and left gonadectomy by laparoscopy were performed. The GCT stage was classified as stage Ic according to FIGO. At this time, pathologic examination showed that the left gonad had progressed to yolk sac tumor and dysgerminoma. The patient underwent chemotherapy post-operatively but developed type III myelosuppression and tumor recurrence several months later. Conclusions The patient initially presented with right gonadoblastoma but chose only right gonadectomy by laparoscopy to preserve the female sex characteristics, which resulted in rapid deterioration of the left gonad and poor treatment outcomes. This case demonstrates the importance of early genetic diagnosis and treatment of 46,XY female DSD.

Sirtuin 1 (Sirt-1) - SRY-Box Transcription Factor 2 (sox2) axis maintains the stemness of human MSCs. Resveratrol may maintain stemness of human iliac bone marrow (BM)-MSCs. The aim of this study to investigate resveratrol effect on sox2 to maintain BM-MSCs stemness through an in silico and in vitro study. BM-MSCs was aspirated from orthopedic patients then, cultured in vitro. The study groups were into a control group, resveratrol group at doses of 0.1 μM and 1 μM. The characterization human iliac BM-MSCs was examined by immunocytochemistry analysis cluster of differentiation (CD)73, CD90, CD105 and CD45. The proliferation of human iliac BM-MSCs in each group was analyzed by MTT assay with various dose of resveratrol 0.01 μM; 0.05 μM; 0.1 μM; 0.5 μM; 1 μM respectively. A molecular docking was done to evaluate the interactions between resveratrol, sirt1 and Sox2 in silico. Resveratrol act as Sirt1 activator with high binding affinity between Sirt1 and Sox2 was -883.9 kcal/mol in silico. BM-MSCs at third, fourth, fifth and sixth sub-cultured with administrated resveratrol at dose 1 μM showed more confluent, less apoptosis and less senescence cells than control group. The characterization of human iliac BM-MSCs at third sub-culture showed that positive expression of CD73, CD90 and CD105 but lack of CD45 expression. There was no significant different of BM-MSCs viability percentage after administration of resveratrol with various doses (p>0.05). Resveratrol has an effect to regulate Sox2 expression that can maintain human illiac BM-MSCs proliferation, self-renewal and stemness in silico and in vitro.

Myocardial fibrosis is the main morphological change of ventricular remodelling caused by cardiovascular diseases, mainly manifested due to the excessive production of collagen proteins. SRY-related high mobility group-box gene 9 (SOX9) is a new target regulating myocardial fibrosis. Bellidifolin (BEL), the active component of G. acuta, can prevent heart damage. However, it is unclear whether BEL can regulate SOX9 to alleviate myocardial fibrosis. The mice were subjected to isoproterenol (ISO) to establish myocardial fibrosis, and human myocardial fibroblasts (HCFs) were activated by TGF-β1 in the present study. The pathological changes of cardiac tissue were observed by HE staining. Masson staining was applied to reveal the collagen deposition in the heart. The measurement for expression of fibrosis-related proteins, SOX9, and TGF-β1 signalling molecules adopted Western blot and immunohistochemistry. The effects of BEL on HCFs, activity were detected by CCK-8. The result showed that BEL did not affect cell viability. And, the data indicated that BEL inhibited the elevations in α-SMA, Collagen I, and Collagen III by decreasing SOX9 expression. Additionally, SOX9 suppression by siRNA downregulated the TGF-β1 expression and prevented Smad3 phosphorylation, as supported by reducing the expression of α-SMA, Collagen I, and Collagen III. In vivo study verified that BEL ameliorated myocardial fibrosis by inhibiting SOX9. Therefore, BEL inhibited SOX9 to block TGF-β1 signalling activation to ameliorate myocardial fibrosis.

Y-encoded transcription factor SRY initiates male differentiation in therian mammals. This factor contains a high-mobility-group (HMG) box, which mediates sequence-specific DNA binding with sharp DNA bending. A companion article in this issue described sex-reversal mutations at box position 72 (residue 127 in human SRY), invariant as Tyr among mammalian orthologs. Although not contacting DNA, the aromatic ring seals the domain’s minor wing at a solvent-exposed junction with a basic tail. A seeming paradox was posed by the native-like biochemical properties of inherited Swyer variant Y72F: its near-native gene-regulatory activity is consistent with the father’s male development, but at odds with the daughter’s XY female somatic phenotype. Surprisingly, aromatic rings (Y72, F72 or W72) confer higher transcriptional activity than do basic or polar side chains generally observed at solvated DNA interfaces (Arg, Lys, His or Gln). Whereas biophysical studies (time-resolved fluorescence resonance energy transfer and heteronuclear NMR spectroscopy) uncovered only subtle perturbations, dissociation of the Y72F complex was markedly accelerated relative to wild-type. Studies of protein-DNA solvation by molecular-dynamics (MD) simulations of an homologous high-resolution crystal structure (SOX18) suggest that Y72 para-OH anchors a network of water molecules at the tail-DNA interface, perturbed in the variant in association with nonlocal conformational fluctuations. Loss of the Y72 anchor among SRY variants presumably “unclamps” its basic tail, leading to (a) rapid DNA dissociation despite native affinity and (b) attenuated transcriptional activity at the edge of sexual ambiguity. Conservation of Y72 suggests that this water-mediated clamp operates generally among SRY and metazoan SOX domains.

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46,XX testicular disorder of sex development (46,XX TDSD) is a relatively rare condition characterised by the presence of testicular tissue with 46,XX karyotype. The present study aims to reveal the phenotype to genotype correlation in a series of sex-determining region Y (SRY)-positive 46,XX TDSD cases. We present the clinical findings, hormone profiles and genetic test results of six patients with SRY-positive 46,XX TDSD and give the details and follow-up findings of our three of previously published patients. All patients presented common characteristics such as azoospermia, hypergonadotropic hypogonadism and an SRY gene translocated on the terminal part of the short arm of one of the X chromosomes. Mean ± standard deviation (SD) height of the patients was 164.78 ± 8.0 cm. Five patients had decreased secondary sexual characteristics, and three patients had gynaecomastia with varying degrees. Five of the seven patients revealed a translocation between protein kinase X (PRKX) and inverted protein kinase Y (PRKY) genes, and the remaining two patients showed a translocation between the pseudoautosomal region 1 (PAR1) of X chromosome and the differential region of Y chromosome. X chromosome inactivation (XCI) analysis results demonstrated random and skewed XCI in 5 cases and 1 case, respectively. In brief, we delineate the phenotypic spectrum of patients with SRY-positive 46,XX TDSD and the underlying mechanisms of Xp;Yp translocations.

Resumen En la mayoría de los casos, la diferenciación sexual masculina ocurre con la participación del gen SRY. Sin embargo, se pueden presentar otros genotipos excepcionales, como en el caso que se presenta en este reporte. Se trata de un paciente adulto de sexo masculino atendido en el Servicio de Paternidades del Instituto de Genética de la Universidad Nacional de Colombia. Se le hicieron los análisis del gen de la amelogenina y de repeticiones cortas en tándem (Short Tandem Repeat, STR) específicas para el gen SRY con estuches comerciales de identificación humana, así como los de cariotipo convencional e hibridación in situ fluorescente del SRY, y el estudio de microdeleciones del cromosoma Y mediante reacción en cadena de la polimerasa (PCR). Se le hizo la evaluación clínica y se le brindó asesoramiento genético. El paciente no presentaba ambigüedad genital, su cariotipo era 46 XX, y el perfil molecular era negativo para el gen SRY y positivo para el ZFY. Se le diagnosticó un trastorno de diferenciación sexual 46 XX testicular no sindrómico, una rara condición genética. Solo el 20 % de los pacientes con este diagnóstico son negativos para SRY y exhiben perfiles moleculares diversos. La información disponible parece indicar que el ZFY está relacionado con la diferenciación sexual masculina, aún en ausencia del gen SRY. Abstract In most cases, male sexual differentiation occurs with SRY gene mediation. However, exceptional genotypes have been identified, as shown in this paper. This was a male adult patient seen at the Servicio de Paternidades, Instituto de Genética, Universidad Nacional de Colombia. The following procedures were carried out: Amelogenin gene and short tandem repeat analyses using human identification commercial kits, conventional karyotype, SRY fluorescent in situ hybridization, PCR analysis for Y chromosome microdeletions, clinical evaluation, and genetic counseling. We present an adult male with unambiguous genitalia, karyotype 46,XX, and an SRY negative and ZFY positive molecular profile. The diagnosis of nonsyndromic 46,XX testicular disorder of sex development (DSD) -a rare genetic condition- was established. Only 20 % of similarly diagnosed patients are SRY negative and exhibit diverse molecular profiles. Until now, available evidence seems to indicate that, even in the absence of SRY, the ZFY factor is involved in male sexual differentiation.

In most cases, male sexual differentiation occurs with SRY gene mediation. However, exceptional genotypes have been identified, as shown in this paper. This was a male adult patient seen at the Servicio de Paternidades, Instituto de Genética, Universidad Nacional de Colombia. The following procedures were carried out: Amelogenin gene and short tandem repeat analyses using human identification commercial kits, conventional karyotype, SRY fluorescent in situ hybridization, PCR analysis for Y chromosome microdeletions, clinical evaluation, and genetic counseling. We present an adult male with unambiguous genitalia, karyotype 46,XX, and an SRY negative and ZFY positive molecular profile. The diagnosis of nonsyndromic 46,XX testicular disorder of sex development (DSD) -a rare genetic condition- was established. Only 20 % of similarly diagnosed patients are SRY negative and exhibit diverse molecular profiles. Until now, available evidence seems to indicate that, even in the absence of SRY, the ZFY factor is involved in male sexual differentiation. ispartof: Biomedica vol:39 issue:4 pages:622-630 ispartof: location:Colombia status: Published online