Sex-determining Region Of Y-chromosome Research Articles

Tolerogenic dendritic cells alleviate collagen-induced arthritis by forming microchimerism and affecting the expression of immune checkpoint molecules.

Tolerogenic dendritic cells (tolDCs) have the potential to treat rheumatoid arthritis (RA) by inducing immune tolerance. However, the mechanism of intervention needs further study. Here, we investigated whether tolDCs formed microchimerism and their effect on the expression of immune checkpoint molecules after infusion of tolDCs into rats with collagen-induced arthritis (CIA). TolDCs derived from male SD rats were labeled with fluorescence and infused into female CIA rats. The fluorescence signals as well as the sex-determining region of Y-chromosome (SRY) gene revealed that tolDCs formed microchimerism in the mesenteric lymph nodes and ankle joints. We further explored the effect of tolDCs on the expression of immune checkpoint molecules in mesenteric lymph nodes and ankle joints. For stimulatory immune checkpoint molecules, the expressions of CD86 and CD40 decreased in mesenteric lymph nodes, and the expressions of CD40, CD40L, CD28, CD80, and CD86 also decreased in rat ankle joints. In contrast, the inhibitory immune checkpoint molecule PDL1 increased in mesenteric lymph nodes, and PD1, PDL1, and CTLA4 increased in ankle joints. In conclusion, our results suggested that intervention of tolDCs in CIA is associated with the formation of microchimerism and the effect on immune checkpoints.

Copy Number Variations of Four Y-Linked Genes in Swamp Buffaloes.

Simple SummaryThe amplification of the male-specific region of the Y chromosome was a unique phenomenon during mammalian sex chromosome evolution. The Y-linked copy number variations of many species have been confirmed. However, the Y-linked copy number variations (CNVs) in water buffalo are still unknown. In this study, we investigated the copy number variations of four Y-linked genes (SRY, UTY, DBY, and OFD1Y) in buffalo. Our results showed that UTY was a single-copy gene in buffalo, while DBY, OFD1Y, and SRY exhibited copy number variations in buffalo.Copy number variation (CNV), a significant source of genetic diversity in the mammalian Y chromosome, is associated with the development of many complex phenotypes, such as spermatogenesis and male fertility. The contribution of Y-linked CNVs has been studied in various species, however, water buffalo has not been explored in this area and the genetic information still remains unknown. The aim of the current study was to investigate the CNVs of four Y-linked genes, including, sex determining Region of Y-Chromosome (SRY), ubiquitously transcribed tetratricopeptide repeat gene protein on the chromosome Y (UTY), DEAD-box helicase 3 Y-linked (DDX3Y, also known as DBY), and oral-facial-digital syndrome 1 Y-linked (OFD1Y) in 254 swamp buffaloes from 15 populations distributed across China, Vietnam, and Laos using quantitative real-time PCR (qPCR). Our results revealed the prevalence of a single-copy UTY gene in buffaloes. The DBY and OFD1Y represented CNVs among and within different buffalo breeds. The SRY showed CNVs only in Vietnamese and Laotian buffaloes. In conclusion, this study indicated that DBY, OFD1Y, and SRY showed CNVs, while the UTY was a single-copy gene in swamp buffaloes.

Clinical and genetic analysis for a patient with 45, X/46, X, Yqh- and mixed gonadal dysgenesis

To investigate the clinical and genetic characteristics of a patient with mixed gonadal dysgenesis. Clinical data was collected. The patient was subjected for serum hormone testing and G-banding chromosomal analysis. Sex-determining region of Y-chromosome (SRY) gene and azoospermia factor (AZF) a, b, c regions were analyzed with multiple polymerase chain reaction (PCR) and whole gene sequencing. All serum hormone testing were normal. The karyotype of the patient was 45,X/46,X,Yqh-. PCR has proven the presence of SRY, ZFY and AZFa, and deletion of AZFb and AZFc regions. No mutation was detected in the sequence of the SRY gene. Abdominal computerized tomography has detected a huge mass in the pelvic cavity, which was positive for PLAP and CD117 on immunohistochemistry stain. Based on clinical data and result of genetic testing, the patient was diagnosed with mixed gonadal dysgenesis. Pathological and immunohistochemistry analysis of the transformed gland has confirmed the diagnosis of seminoma. For patient with a karyotype of 45,X/46,X,Yqh-, the risk of seminoma may be related with the presence of SRY gene.

Clinical characteristics and molecular genetics of six patients with Swyer syndrome

Objective To analyze the clinical data of six patients with Swyer syndrome and to explore the genetic defects in this disease.Methods The clinical features and laboratory data of six patients were collected.The exon or fragments of sex-determining region of Y-chromosome (SRY) gene were amplified by PCR.Results All six patients had 46,XY karyotype,presenting with primary amenorrhea,female infant vulva,tiny uterus without testis or prostate.Ovarian or streak gonads were detected in three of them.Sequencing analysis showed that the missense mutations,p.R62G,p.R76L,and p.S151 * were identified in three patients respectively,and there were a large fragment deletion in one patient and no mutation or deletion in two patients.The mutations of p.R76L and p.S151 *,and the large fragment deletion of SRY gene were not previously reported.Conclusion SRY mutation and deletion are the main causes of Swyer syndrome.The patients with Swyer syndrome should be screened for SRY gene mutations by priority. Key words: Swyer syndrome ; SRY gene ; Mutation ; Deletion

SRY gene analysis for a case with sex reversal syndrome

To investigate the molecular mechanism of sex reversal in a 46,XY female patient. Clinical data was collected. Peripheral blood lymphocytes were cultured for G-banding chromosomal analysis and DNA extraction. Sex-determining region of Y-chromosome (SRY) gene was analyzed with polymerase chain reaction (PCR) and DNA sequencing . Although the patient has a female appearance, he has a karyotype of 46,XY. The SRY gene can be detected in all samples. The 6th base of SRY gene coding region was deleted, resulting in a frameshifting mutation and premature termination of protein translation. The sex reversal of the patient is probably due to abnormal embryonic development caused by the SRY gene mutation.

Bone marrow mesenchymal stromal cells with support of bispecific antibody and ultrasound-mediated microbubbles prevent myocardial fibrosis via the signal transducer and activators of transcription signaling pathway

This study was initiated to investigate the efficacy of myocardial fibrosis intervention via signal transducer and activators of transcription (STAT) signaling using bone marrow (BM) mesenchymal stromal cells (MSC) with the aid of bispecific antibody (BiAb) and ultrasound-mediated microbubbles (MB). BiAb (anti-CD29 × anti-myosin light chain antibody; AMLCA) was prepared and combined with isolated MSC from male mice and transfused into female mice with isoproterenol-induced myocardial fibrosis via the tail vein, followed by MB (MSC + BiAb + MB). This study included seven groups: MSC + BiAb + MB; MSC; BiAb; MB; MSC + BiAb; untreated; and control. Five weeks after treatment, expression levels of the sex-determining region of Y-chromosome (SRY), matrix metalloproteinases (MMP)-9, tissue inhibitor of metalloproteinase (TIMP)-1 and vascular endothelial growth factor (VEGF) in myocardium were detected by fluorescent quantitative real-time polymerase chain reaction (qRT-PCR). Collagen distribution was observed using Sirius Red staining. The protein expression of signal transducer and activators of transcription (STAT)1 and STAT3 was detected by Western blot. The highest homing number of MSC was in the MSC + BiAb + MB group, second highest in the MSC + BiAb group, and lowest in MSC alone. Compared with the untreated group, MSC + BiAb + MB, MSC + BiAb and MSC groups had decreased levels of MMP-9, TIMP-1, STAT1 and collagen deposition, and increased levels of STAT3. Upregulated STAT3 and downregulated TIMP-1 were significantly different in MSC + BiAb + MB compared with MSC alone or MSC + BiAb. The homing rate and repairing efficacy of MSC improved with treatment utilizing a combination of BiAb and MB. MSC can improve MMP-TIMP expression in injured myocardium and interfere with myocardial fibrosis after homing, a mechanism that may be related to the STAT-mediated signaling pathway.

Bone marrow mesenchymal stem cells with support of ultrasound-mediated microbubbles and bispecific antibody prevent myocardial fibrosis via HSP-70

Objective To explore the effects of transplantated bone marrow mesenchymal stem cells (MSCs) on myocardial fibrosis with the aid of ultrasound-mediated microbubbles (MB) and bispecific antibody(BiAb) combination.Methods With the aid of MB,isolated MSCs from male mice and the BiAb were transfused into female mice with isoproterenol-induced myocardial fibrosis via tail vein (MSCs + BiAb + MB group).BiAb was producted with anti-CD29 which can recognize MSCs and anti-myosin light chain antibody (AMLCA) which can specifically bind to injured myocardium.There were six groups investigated:MSC + BiAb + MB,MSC,BiAb,MB,MSC + BiAb,untreated,and control groups.Five weeks after treatment administration,the expressions of sex-determining region of Y-chromosome (SRY) in myocardium were detected by fluorescent quantitative PCR.The distribution of collagen was observed by sirius red staining.Heat shock protein-70 (HSP-70) in myocardium was detected by immunohistochemistry.Results The highest homing number of MSCs was in the MSCs + BiAb + MB group,MSCs + BiAb group ranked the second,and lowest in MSCs group.Compared to the untreated group,the MSCs + BiAb + MB,MSCs + BiAb and MSCs groups had less collagen deposition (P ＜0.05),and decreased level of HSP-70.Compared to those of the MSCs group,the collagen deposition were decreased in MSCs + BiAb + MB group (P ＜0.05).Conclusions MB and BiAb can promote the homing number of MSCs in mice.MB can further the homing rate and the repairing efficacy of MSCs.The homing MSCs can prevent the process of myocardial fibrosis.And HSP-70 was involved in the internal mechanism. Key words: Ultrasonography; Microbubbles; Mesenchymal stem cell ; Antibody,bispecific ; Myocardial fibrosis

INCIDENCE OF MICRODELETIONS IN THE AZF REGION OF THE Y CHROMOSOME IN SLOVAK PATIENTS WITH AZOOSPERMIA

The Y chromosome accumulates male-related genes including sex-determining region of Y-chromosome (SRY) and several spermatogenesis-related genes. The long arm contains azoospermia factor (AZF) region (including sub-regions AZFa, AZFb and AZFc). Microdeletions in this region are responsible for azoospermia and oligospermia and result in the male infertility. The aim of this study was to analyze incidence of microdeletions in the AZF region of Y chromosome in patients with azoospermia from Slovakia. Over the period from 2005 to 2009 a total of 239 men (mean age 31.74 years) were analyzed. The diagnosis of azoospermia was established on the basis of semen analysis. All patient samples were analyzed cytogenetically. Chromosomal analysis was performed on all patients on cultured lymphocytes from peripheral blood. For exact diagnosis of microdeletions in AZF region we used a PCR-method using a set of sequence-tagged sites from all AZF sub-regions (according to the recommendation by the European Academy of Andrology and the European Quality Monitoring Network Group). Among our 226 patients with azoospermia and with normal karyotype, 8 patients (mean age 30.6 years) had microdeletions in the AZF region of the Y chromosome (3.35%). Considering particular types of deletions we determined deletions in each region AZFa,b,c but also a complete deletion of the entire AZF region. The presence of microdeletion(s) in the AZFc region was the most frequent. In our study we found 12 patients (5%) with 47,XXY karyotype (Klinefelter syndrome), but these patients didn't have microdeletion of Y chromosomes. The study confirmed that percentage of microdeletions in the AZF region is low in Slovak azoospermic patients, but important from a prognostic view.

Biocompatibility of acellular corneal stroma and transplantation of tissue-engineered corneal epithelium

To evaluate the biocompatibility of xenogeneic acellular corneal stroma, the feasibility of tissue engineered corneal epithelial transplantation, and verify the long term survival of epithelial allograft. It was a experimental study. Porcine corneal stroma was treated by dispase followed by Triton-X-100 detergent. Treated porcine corneal stroma (group A) or fresh corneal stroma (group B) were put into the sac of rabbit cornea. Rabbit cornea without implantation of porcine corneal stroma was used as the control group (group C). Immunological rejection was evaluated in morphology, histopathology and immunohistochemistry in month 1, 3, 6. Female rabbit underwent lamellar keratoplasty (LK) using male tissue engineered corneal epithelium as donors, and immunological rejection after LK was analyzed. The corneas were collected at day 1, 3, 7, 15 and 30 after LK and evaluated by histopathology, immunohistochemistry and sex-determining region of Y-chromosome (SRY)-PCR analysis. All corneas became transparent gradually after the transplantation of treated porcine corneal stroma and were not rejected. The epithelium, stroma, endothelium, Bowman's and Descemet's membrane were preserved in all corneas of group A and B in histological observations, collagen fibers were parallel, a few keratocytes presented in the acellular and fresh corneal stromas. The corneas of group C were normal in histological sections. No significant immune rejection was noted in any of the corneas. The corneas in the study of transplantation of tissue engineered cornea epithelium recovered smoothly in 3 or 4 days, turned transparent in 15 or 20 days after surgery and were not stained by fluorescein. Well-differentiated corneal epithelium were recognized at 15 and 30 days after LK. Many keratocytes infiltrated into the scaffold. SRY-PCR analysis showed that allogenic donor corneal epithelium cells could survive in recipients after a long-term observation. Acellular porcine corneal stroma shows a satisfying biocompatibility. Tissue engineered corneal epithelium using acellular porcine corneal stroma as carrier could be used as donors for LK with satisfactory results. Donor cells have the potential to survive in recipients after long-term observation.

“Micro-deletions” of the human Y chromosome and their relationship with male infertility

The Y chromosome evolves from an autochromosome and accumulates male-related genes including sex-determining region of Y-chromosome (SRY) and several spermatogenesis-related genes. The human Y chromosome (60 Mb long) is largely composed of repetitive sequences that give it a heterochromatic appearance, and it consists of pseudoautosomal, euchromatic, and heterochromatic regions. Located on the two extremities of the Y chromosome, pseudoautosomal regions 1 and 2 (PAR1 and PAR2, 2.6 Mb and 320 bp long, respectively) are homologs with the termini of the X chromosome. The euchromatic region and some of the repeat-rich heterochromatic parts of the Y chromosome are called “male-specific Y” (MSY), which occupy more than 95% of the whole Y chromosome. After evolution, the Y chromosome becomes the smallest in size with the least number of genes but with the most number of copies of genes that are mostly spermatogenesis-related. The Y chromosome is characterized by highly repetitive sequences (including direct repeats, inverted repeats, and palindromes) and high polymorphism. Several gene rearrangements on the Y chromosome occur during evolution owing to its specific gene structure. The consequences of such rearrangements are not only loss but also gain of specific genes. One hundred and fifty three haplotypes have been discovered in the human Y chromosome. The structure of the Y chromosome in the GenBank belongs to haplotype R1. There are 220 genes (104 coding genes, 111 pseudogenes, and 5 other uncategorized genes) according to the most recent count. The 104 coding genes encode a total of about 48 proteins/protein families (including putative proteins/protein families). Among them, 16 gene products have been discovered in the azoospermia factor region (AZF) and are related to spermatogenesis. It has been discovered that one subset of gene rearrangements on the Y chromosome, “micro-deletions”, is a major cause of male infertility in some populations. However, controversies exist about different Y chromosome haplotypes. Six AZFs of the Y chromosome have been discovered including AZFa, AZFb, AZFc, and their combinations AZFbc, AZFabc, and partial AZFc called AZFc/gr/gr. Different deletions in AZF lead to different content spermatogenesis loss from teratozoospermia to infertility in different populations depending on their Y haplotypes. This article describes the structure of the human Y chromosome and investigates the causes of micro-deletions and their relationship with male infertility from the view of chromosome evolution. After analysis of the relationship between AZFc and male infertility, we concluded that spermatogenesis is controlled by a network of genes, which may locate on the Y chromosome, the autochromosomes, or even on the X chromosome. Further investigation of the molecular mechanisms underlying male fertility/infertility will facilitate our knowledge of functional genomics.

The fate of donor bone cells in different diameter bone grafts: an experiment study

To investigate the fate of donor bone cells in bone grafts of different diameters during repairing bone defects. One hundred sixteen female syngeneic inbred DA rats were established as radial defect model of 3 kinds: structural bone grafting group (n = 56), morselized bone grafting group (n = 56), and blank group (without bone grafting, n = 4), to be used as receptors The ilia of 58 male inbred DA rats, as donors, were harvested and made into structural bone grafts 2 mm in diameter and morselized bone grafts 0.3 - 0.5 mm in diameter to be transplanted into the radial defects of the female receptors. One and four days, and 1, 2, 4, 6, and 8 weeks after transplantation DNA was extracted from the grafted bones and polymerase chain reaction (PCR) specific for the sex-determining region of Y-chromosome (Sry) was performed to observe the presence and relative amount of Y-chromosome originating from the bone grafts, expression of the sex-determining gene Sry in the receptors' bones and the histology of the receptors' bones. In structural bone grafting group, the amounts of Sry-specific bands decreased in the early time and disappeared 1 week after transplantation, and re-appeared 4 weeks after transplantation with the amount increasing with the lapse of time. In morselized bone grafting group, Sry-specific bands were detected all the time but their amounts decreased with the lapse of time. At each time point, morselized bone graft provided more living osteocytes with better effect of osteogenesis in comparison with the structural bone graft. Bone grafts of different diameters provide donor cells in repairing bone defects. Having more surviving osteocytes, morselized bone grafts may accelerate the healing pf bone defects, thus providing a new and effective method to repair bone defects and spinal fusion clinically.