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1.
Am J Hum Genet ; 108(2): 324-336, 2021 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-33508233

RESUMO

Human infertility is a multifactorial disease that affects 8%-12% of reproductive-aged couples worldwide. However, the genetic causes of human infertility are still poorly understood. Synaptonemal complex (SC) is a conserved tripartite structure that holds homologous chromosomes together and plays an indispensable role in the meiotic progression. Here, we identified three homozygous mutations in the SC coding gene C14orf39/SIX6OS1 in infertile individuals from different ethnic populations by whole-exome sequencing (WES). These mutations include a frameshift mutation (c.204_205del [p.His68Glnfs∗2]) from a consanguineous Pakistani family with two males suffering from non-obstructive azoospermia (NOA) and one female diagnosed with premature ovarian insufficiency (POI) as well as a nonsense mutation (c.958G>T [p.Glu320∗]) and a splicing mutation (c.1180-3C>G) in two unrelated Chinese men (individual P3907 and individual P6032, respectively) with meiotic arrest. Mutations in C14orf39 resulted in truncated proteins that retained SYCE1 binding but exhibited impaired polycomplex formation between C14ORF39 and SYCE1. Further cytological analyses of meiosis in germ cells revealed that the affected familial males with the C14orf39 frameshift mutation displayed complete asynapsis between homologous chromosomes, while the affected Chinese men carrying the nonsense or splicing mutation showed incomplete synapsis. The phenotypes of NOA and POI in affected individuals were well recapitulated by Six6os1 mutant mice carrying an analogous mutation. Collectively, our findings in humans and mice highlight the conserved role of C14ORF39/SIX6OS1 in SC assembly and indicate that the homozygous mutations in C14orf39/SIX6OS1 described here are responsible for infertility of these affected individuals, thus expanding our understanding of the genetic basis of human infertility.


Assuntos
Azoospermia/genética , Mutação , Insuficiência Ovariana Primária/genética , Adulto , Azoospermia/fisiopatologia , Pareamento Cromossômico , Códon sem Sentido , Proteínas de Ligação a DNA/metabolismo , Feminino , Homozigoto , Humanos , Masculino , Meiose , Pessoa de Meia-Idade , Proteínas Nucleares/metabolismo , Linhagem , Insuficiência Ovariana Primária/fisiopatologia , Espermatócitos/metabolismo , Espermatócitos/fisiologia , Complexo Sinaptonêmico/genética , Complexo Sinaptonêmico/metabolismo , Sequenciamento Completo do Genoma
2.
Gene ; 765: 145045, 2021 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-32777524

RESUMO

To find the variant spectrum of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, and evaluate its frequent variants in Chinese congenital absence of vas deferens (CAVD) patients. A total of 276 patients with azoospermia and CAVD (aged from 21 to 44 years old) were investigated from May 2013 to September 2019 in the Third Affiliated Hospital of Sun Yat-sen University. Additionally, 50 healthy, unrelated volunteers were recruited as controls (aged from 21 to 46 years old). The 5'-UTR, exons and their flanking side of the CFTR gene were sequenced by high-throughput sequencing technology. The results were compared with those retrieved from the Ensembl Genome Browser. In addition, all 13 novel variants were further confirmed independently by Sanger sequencing and evaluated in the bioinformatics web servers. A schematic of the variant spectrum of the CFTR gene, including 13 novel variants (12 in CAVD patients, one in the control group), is shown, and the frequent variants in Chinese CAVD patients were 5 T (27.54%), c.-8G > C (7.25%), p.Q1352H (5.98%), and p.I556V (3.08%). 5 T was found to be the most frequent variant. p.Q1352H had a significantly high allelic frequency in CAVD patients (P < 0.05). c.-8G > C and p.I556V had high allelic frequencies but showed no difference between patients and controls (P > 0.05). p.Q1352H is the most common and important missense variant in Chinese patients with CAVD, while the pathological effects of C.-8G > C and p.I556V may be weak after evaluation.


Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/genética , Doenças Urogenitais Masculinas/genética , Ducto Deferente/anormalidades , Adulto , Alelos , Grupo com Ancestrais do Continente Asiático/genética , Azoospermia/genética , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Análise Mutacional de DNA/métodos , Éxons/genética , Frequência do Gene/genética , Humanos , Infertilidade Masculina/genética , Masculino , Doenças Urogenitais Masculinas/metabolismo , Mutação/genética , Ducto Deferente/metabolismo
3.
Zhonghua Nan Ke Xue ; 26(2): 149-153, 2020 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-33346419

RESUMO

Objective: To explore the clinical characteristics and prognosis of the 48,XXYY syndrome and gain a deeper insight into this condition. METHODS: This retrospective study included 4 cases of 48,XXYY syndrome confirmed between 2011 and 2018. We analyzed the general information, clinical manifestations, laboratory results, imaging features and outcomes of assisted reproductive technology (ART) of the patients and reviewed the relevant literature. RESULTS: The 4 patients with 48,XXYY syndrome were characterized by low literacy, soft texture and small volume of the testis, high levels of FSH and LH, and low level of serum T. Two of them were diagnosed with ejaculatory dysfunction and aspermia, and the other 2 with normal ejaculatory function but azoospermia. Biochemical analysis of seminal plasma indicated normal quantifications of both fructose and neutral α glucosidase. ART with donor sperm was performed for all the 4 cases and 3 of them got full-term babies. CONCLUSIONS: The 48,XXYY syndrome is often complicated by hypergonadotropic hypogonadism, with the clinical manifestations of aspermia or non-obstructive azoospermia. ART with donor sperm can be employed to help the patient with 48,XXYY syndrome get their non-biological offspring.


Assuntos
Transtornos dos Cromossomos Sexuais/diagnóstico , Azoospermia/genética , Humanos , Masculino , Técnicas de Reprodução Assistida , Estudos Retrospectivos , Sêmen/química , Transtornos dos Cromossomos Sexuais/patologia , Testículo
4.
Medicine (Baltimore) ; 99(31): e21107, 2020 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-32756090

RESUMO

RATIONALE: Infertility is a common medical condition that affects nearly 15% of the world population. Non-obstructive azoospermia (NOA) is one of the most severe forms of male infertility. Some common structural variants, single nucleotide polymorphisms (SNPs), and genetic factors were reported to be associated with NOA. However, the underlying etiology and genetic mechanism(s) remain largely unclear. This report aimed to describe the associated mutation of the heat shock factor-2 (HSF2) gene in Chinese infertile men with NOA. PATIENT CONCERNS: An apparently healthy 27-year-old man with a body mass index (BMI) of 23.31 kg/m had a 2-year history of primary infertility. DIAGNOSES: The semen analysis of the patient showed a sperm concentration of 0/mL in 6.5 mL of semen. The patient was diagnosed with NOA by performing the comprehensive examinations including a detailed medical history, physical examination, chromosome analysis, Y-chromosome microdeletions, semen analysis, and hormone profiles. INTERVENTIONS: The couple received artificial insemination by donor (AID) and a healthy girl was born after the embryo transfer. OUTCOMES: We found a novel deletion-insertion variation c.326_326delinsGGAAGGTGAGCTATTGT in the exon 3 of the HSF2 gene by performing next-generation sequencing on him who was diagnosed NOA. We performed Sanger sequencing on this patient and confirmed the heterozygous missing insertion mutation in the patient. This is a novel mutation. The variant was heterozygous and categorized as pathogenic. LESSONS: A novel deletion-insertion variation c.326_326delinsGGAAGGTGAGCTATTGT in the exon 3 of HSF2 gene HSF2 is predicted to be pathogenic and associated with the occurrence of NOA.


Assuntos
Azoospermia/genética , Proteínas de Choque Térmico/genética , Mutação INDEL/genética , Fatores de Transcrição/genética , Adulto , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Análise do Sêmen
5.
Am J Hum Genet ; 107(2): 342-351, 2020 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-32673564

RESUMO

Male infertility affects ∼7% of men, but its causes remain poorly understood. The most severe form is non-obstructive azoospermia (NOA), which is, in part, caused by an arrest at meiosis. So far, only a few validated disease-associated genes have been reported. To address this gap, we performed whole-exome sequencing in 58 men with unexplained meiotic arrest and identified the same homozygous frameshift variant c.676dup (p.Trp226LeufsTer4) in M1AP, encoding meiosis 1 associated protein, in three unrelated men. This variant most likely results in a truncated protein as shown in vitro by heterologous expression of mutant M1AP. Next, we screened four large cohorts of infertile men and identified three additional individuals carrying homozygous c.676dup and three carrying combinations of this and other likely causal variants in M1AP. Moreover, a homozygous missense variant, c.1166C>T (p.Pro389Leu), segregated with infertility in five men from a consanguineous Turkish family. The common phenotype between all affected men was NOA, but occasionally spermatids and rarely a few spermatozoa in the semen were observed. A similar phenotype has been described for mice with disruption of M1ap. Collectively, these findings demonstrate that mutations in M1AP are a relatively frequent cause of autosomal recessive severe spermatogenic failure and male infertility with strong clinical validity.


Assuntos
Pontos de Checagem do Ciclo Celular/genética , Infertilidade Masculina/genética , Meiose/genética , Mutação/genética , Proteínas/genética , Espermatogênese/genética , Adulto , Alelos , Animais , Azoospermia/genética , Homozigoto , Humanos , Masculino , Camundongos , Fenótipo , Espermatozoides/anormalidades , Testículo/anormalidades , Turquia , Sequenciamento Completo do Exoma/métodos
6.
Medicine (Baltimore) ; 99(23): e20561, 2020 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-32502024

RESUMO

Nonobstructive azoospermia (NOA) is a severe form of male infertility. The molecular basis of NOA is still poorly understood. The aim of this study was to explore the associations between single nucleotide polymorphisms (SNPs) of the TATA-box binding protein associated factor 4b (TAF4B) gene and NOA. A total of 100 Han Chinese patients with NOA and 100 healthy men as controls were recruited. Targeted gene capture sequencing was performed. A total of 11 TAF4B SNPs were screened in the NOA and control subjects. Six synonymous and 4 nonsynonymous variants were detected. The c.11G>T (p.G4V) mutation was detected only in NOA patients. Polymorphism Phenotyping v2 and Sorting Intolerant From Tolerant analysis indicated that the p.G4V mutation influenced the protein structure of TAF4B. Haplotype analysis showed that the candidate SNPs did not independently associate with NOA and were found at extremely low frequencies in the subject population. Mutation Taster analysis indicated that the c.11G>T/p.G4V mutation was damaging. WebLogo analysis showed that the residue at amino acid 4 was relatively conserved. The p.Gly4Val substitution may affect the structure of the TAF4B protein. The c.11G>T mutation of the TAF4B gene may be associated with NOA in a Chinese population. Bioinformatics analysis indicated this variation may play an important role in the process of spermatogenesis.


Assuntos
Azoospermia/genética , Fatores Associados à Proteína de Ligação a TATA/genética , Fator de Transcrição TFIID/genética , Adulto , Estudos de Casos e Controles , China , Humanos , Masculino , Mutação , Polimorfismo de Nucleotídeo Único , Adulto Jovem
7.
Sci China Life Sci ; 63(7): 1006-1015, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32361911

RESUMO

Being infected by SARS-CoV-2 may cause damage to multiple organs in patients, such as the lung, liver and heart. Angiotensin-converting enzyme 2 (ACE2), reported as a SARS-CoV-2 receptor, is also expressed in human male testes. This suggests a potential risk in human male reproductive system. However, the characteristics of ACE2-positive cells and the expression of other SARS-CoV-2 process-related genes are still worthy of further investigation. Here, we performed singlecell RNA seq (scRNA-seq) analysis on 853 male embryo primordial germ cells (PGCs) and 2,854 normal testis cells to assess the effects of the SARS-CoV-2 virus on the male reproductive system from embryonic stage to adulthood. We also collected and constructed the scRNA-seq library on 228 Sertoli cells from three non-obstructive azoospermia (NOA) patients to assess the effects at disease state. We found that ACE2 expressing cells existed in almost all testis cell types and Sertoli cells had highest expression level and positive cells ratio. Moreover, ACE2 was also expressed in human male PGCs. In adulthood, the level of ACE2 expression decreased with the increase of age. We also found that ACE2 positive cells had high expressions of stress response and immune activation-related genes. Interestingly, some potential SARS-CoV-2 process-related genes such as TMPRSS2, BSG, CTSL and CTSB had different expression patterns in the same cell type. Furthermore, ACE2 expression level in NOA donors' Sertoli cells was significantly decreased. Our work would help to assess the risk of SARS-CoV-2 infection in the male reproductive system.


Assuntos
Azoospermia/genética , Betacoronavirus/patogenicidade , Infecções por Coronavirus , Pandemias , Peptidil Dipeptidase A/genética , Pneumonia Viral , Testículo/metabolismo , Testículo/virologia , Adulto , Azoospermia/complicações , Azoospermia/metabolismo , Betacoronavirus/metabolismo , Infecções por Coronavirus/complicações , Infecções por Coronavirus/fisiopatologia , Infecções por Coronavirus/virologia , Células Germinativas Embrionárias/metabolismo , Células Germinativas Embrionárias/virologia , Expressão Gênica , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Masculino , Peptidil Dipeptidase A/metabolismo , Pneumonia Viral/complicações , Pneumonia Viral/fisiopatologia , Pneumonia Viral/virologia , Receptores Virais/genética , Receptores Virais/metabolismo , Células de Sertoli/metabolismo , Células de Sertoli/virologia , Análise de Célula Única , Espermatogênese/genética , Espermatogênese/fisiologia , Testículo/citologia
8.
Urol Clin North Am ; 47(2): 147-155, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32272986

RESUMO

For men with obstructive azoospermia, several surgical sperm retrieval techniques can facilitate conception with assisted reproductive technology. The evolution of both percutaneous and open approaches to sperm retrieval has been affected by technological innovations, including the surgical microscope, in vitro fertilization, and intracytoplasmic sperm injection. Further modifications to these procedures are designed to minimize patient morbidity and increase the quality and quantity of sperm samples. Innovative technologies promise to further ameliorate outcomes by selecting the highest quality sperm. Although various approaches to surgical sperm retrieval are now well established, several advancements in sperm selection and optimization are being developed.


Assuntos
Azoospermia/diagnóstico , Azoospermia/genética , Fertilização In Vitro/métodos , Injeções de Esperma Intracitoplásmicas/métodos , Recuperação Espermática , Fragmentação do DNA , Humanos , Masculino , Análise do Sêmen
9.
Urol Clin North Am ; 47(2): 157-164, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32272987

RESUMO

Guiding a couple with nonobstructive azoospermia requires an integrated approach to care by the urologist and the reproductive endocrinologist. After informing the couple of the implications of the diagnosis, care must be taken to outline the options of parenthood. Most experts agree that sperm retrieval in men can be challenging. This article describes various options of sperm retrieval, historic and contemporary, and highlights the advantages and disadvantages of each. The authors find that using a testicular map can invariably help guide sperm retrieval and overall fertility care. The right approach is one that involves a shared decision with the couple.


Assuntos
Azoospermia/diagnóstico , Azoospermia/genética , Biópsia por Agulha Fina/métodos , Recuperação Espermática , Testículo/patologia , Azoospermia/etiologia , Humanos , Masculino , Microdissecção , Análise do Sêmen , Recuperação Espermática/efeitos adversos , Espermatozoides/patologia
10.
Fertil Steril ; 113(3): 561-568, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32111475

RESUMO

OBJECTIVE: To identify the genetic cause of male factor infertility characterized by severe oligozoospermia. DESIGN: Genetic studies. SETTING: Medical university. PATIENT(S): Two infertile brothers with severe oligozoospermia in a consanguineous Han Chinese family, 414 additional patients with oligo-/azoospermia, and 223 fertile (control) subjects. INVENTION(S): None. MAIN OUTCOME MEASURE(S): Genetic analyses using whole-exome and Sanger sequencing were performed for two brothers with severe oligozoospermia. The effects of an identified candidate causative mutation were investigated in silico and in vitro. Whole-exome sequencing screening for the candidate mutation was conducted in 414 patients with oligo-/azoospermia and 223 fertile subjects. RESULT(S): A homozygous missense variant (NM_080746:c.A257C: p.H86P) in RPL10L was identified in the two affected brothers and shown to cosegregate with the severe oligozoospermia phenotype. The mutation was absent in public databases, including the 1000 Genomes Project and the Exome Aggregation Consortium. All queried databases predicted the mutation to be damaging, consistent with the fact that it decreased protein levels in vitro. Subsequent mutation screening identified three additional heterozygous RPL10L mutations in three of 414 subjects with oligo-/azoospermia, but no RPL10L mutations among 223 fertile subjects. CONCLUSION(S): Our findings implicate RPL10L as a novel candidate gene in the pathogenesis of human male factor infertility and severe oligozoospermia.


Assuntos
Azoospermia/genética , Infertilidade Masculina/genética , Mutação de Sentido Incorreto , Oligospermia/genética , Proteínas Ribossômicas/genética , Adulto , Grupo com Ancestrais do Continente Asiático/genética , Azoospermia/complicações , Estudos de Casos e Controles , China , Consanguinidade , Frequência do Gene , Testes Genéticos , Homozigoto , Humanos , Infertilidade Masculina/etiologia , Masculino , Oligospermia/complicações , Linhagem , Irmãos , Sequenciamento Completo do Exoma
11.
Gene ; 737: 144481, 2020 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-32070749

RESUMO

Studies have recently demonstrated that mesenchymal stem cells (MSCs) have therapeutic capabilities on many diseases and this effect is mainly mediated by miRNAs. However, the actual mechanism of MSCs paracrine effect on testis to improve male fertility is still elusive. Herein, we evaluated the altered expression of some spermatogenesis-related miRNAs and their target genes following transplantation of bone marrow (BM)-derived MSCs into testes of busulfan-induced azoospermic rats using real time PCR. Transplantation of MSCs improved fertility of azoospermic rats as revealed by enhanced serum levels of testosterone and estradiol, and upregulated expression of germ cell­specific genes. Azo rats injected with MSCs also exhibited a significant downregulated expression of miRNA-19b, miRNA-100, miRNA-141, miRNA­146a, miRNA-429, and let­7a and a significant upregulated expression of miRNA-21, miRNA-34b, miRNA-34c, miRNA-122, miRNA-449a, miRNA-449b, and miRNA-449c in the testis as compared to Azo rats injected with phosphate buffer saline. Transplantation of MSCs was also accompanied with restoration of the disrupted expression of Ccnd1, E2F1, Myc, and PLCXD3 (target genes for miRNA-34 and miRNA­449 clusters) and ERα and AKT1 (target genes for miRNA-100 and let­7a) to level comparable to that of the fertile group. Upon these data, we infer that BM-MSCs can improve fertility of azoospermic rats and this effect was followed by altered expression of some spermatogenesis-related miRNAs and their target genes. These findings provide MSCs as a promising and effective cell-based therapeutic method for azoospermic patients, but further investigations are required before clinical application.


Assuntos
Azoospermia/induzido quimicamente , Bussulfano/toxicidade , Células-Tronco Mesenquimais/efeitos dos fármacos , MicroRNAs/genética , Animais , Azoospermia/genética , Azoospermia/fisiopatologia , Regulação para Baixo , Feminino , Fertilidade , Masculino , Células-Tronco Mesenquimais/metabolismo , Ratos , Espermatogênese/genética
12.
BMC Med Genet ; 21(1): 33, 2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-32059713

RESUMO

BACKGROUND: Tudor domain-containing proteins (TDRDs) play a critical role in piRNA biogenesis and germ cell development. piRNAs, small regulatory RNAs, act by silencing of transposons during germline development and it has recently been shown in animal model studies that defects in TDRD genes can lead to sterility in males. METHODS: Here we evaluate gene and protein expression levels of four key TDRDs (TDRD1, TDRD5, TDRD9 and TDRD12) in testicular biopsy samples obtained from men with obstructive azoospermia (OA, n = 29), as controls, and various types of non-obstructive azoospermia containing hypospermatogenesis (HP, 28), maturation arrest (MA, n = 30), and Sertoli cell-only syndrome (SCOS, n = 32) as cases. One-way ANOVA test followed by Dunnett's multiple comparison post-test was used to determine inter-group differences in TDRD gene expression among cases and controls. RESULTS: The results showed very low expression of TDRD genes in SCOS specimens. Also, the expression of TDRD1 and TDRD9 genes were lower in MA samples compared to OA samples. The expression of TDRD5 significantly reduced in SCOS, MA and HP specimens than the OA specimens. Indeed, TDRD12 exhibited a very low expression in HP specimens in comparison to OA specimens. All these results were confirmed by Western blot technique. CONCLUSION: TDRDs could be very important in male infertility, which should be express in certain stages of spermatogenesis.


Assuntos
Azoospermia/genética , Proteínas de Ciclo Celular/genética , DNA Helicases/genética , Infertilidade Masculina/genética , Adulto , Animais , Azoospermia/patologia , Regulação da Expressão Gênica/genética , Humanos , Infertilidade Masculina/patologia , Masculino , RNA Interferente Pequeno/genética , Espermatogênese/genética , Testículo/crescimento & desenvolvimento , Testículo/metabolismo , Testículo/patologia
13.
PLoS One ; 15(2): e0229503, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32092127

RESUMO

BACKGROUND: Cattleyak are the hybrid offspring between cattle and yak and combine yak hardiness with cattle productivity. Much attempt has been made to examine the mechanisms of male sterility caused by spermatogenic arrest, but yet there is no research systematically and precisely elucidated testis gene expression profiling between cattleyak and yak. METHODS: To explore the higher resolution comparative transcriptome map between the testes of yak and cattleyak, and further analyze the mRNA expression dynamics of spermatogenic arrest in cattleyak. We characterized the comparative transcriptome profile from the testes of yak and cattleyak using high-throughput sequencing. Then we used quantitative analysis to validate several differentially expressed genes (DEGs) in testicular tissue and spermatogenic cells. RESULTS: Testis transcriptome profiling identified 6477 DEGs (2919 upregulated and 3558 downregulated) between cattleyak and yak. Further analysis revealed that the marker genes and apoptosis regulatory genes for undifferentiated spermatogonia were upregulated, while the genes for differentiation maintenance were downregulated in cattleyak. A majority of DEGs associated with mitotic checkpoint, and cell cycle progression were downregulated in cattleyak during spermatogonial mitosis. Furthermore, almost all DEGs related to synaptonemal complex assembly, and meiotic progression presented no sign of expression in cattleyak. Even worse, dozens of genes involved in acrosome formation, and flagellar development were dominantly downregulated in cattleyak. CONCLUSION: DEGs indicated that spermatogenic arrest of cattleyak may originate from the differentiation stage of spermatogonial stem cells and be aggravated during spermatogonial mitosis and spermatocyte meiosis, which contributes to the scarcely presented sperms in cattleyak.


Assuntos
Azoospermia/congênito , Quimera/genética , Infertilidade Masculina/genética , Animais , Azoospermia/genética , Bovinos/genética , Perfilação da Expressão Gênica/métodos , Perfilação da Expressão Gênica/veterinária , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Masculino , Meiose/genética , Espermatócitos/metabolismo , Espermatogênese/genética , Espermatogônias/metabolismo , Espermatozoides/metabolismo , Testículo/metabolismo , Transcriptoma/genética
14.
Gene ; 735: 144389, 2020 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-31982552

RESUMO

Azoospermia factors, located in the long arm of the Y chromosome, are critical for spermatogenesis, the microdeletions of AZF are considered to be associated with male infertility. In addition to complete deletion, several AZFc partial deletions were also detected in infertile men with wide phenotypic heterogeneity. In this study, we investigated the relevance of Y chromosome deletions, Y-linked CNVs and variable phenotypes in infertile men. To clarify the relationship between phenotypic heterogeneity and Y chromosome deletion in male infertility, we performed chromosomal microarray analysis (CMA) capable of analyzing thousands of loci simultaneously to investigate Y-linked copy number variations (CNVs). Firstly, we reviewed the results of Y chromosome screening in 554 infertile patients and then compared the results of CMA to routine Y chromosome screening in 29 patients with Y chromosomal microdeletions. Then, the Y-linked CNVs associated with oligoasthenospermia were identified according to ACMG standards and guidelines. The results indicated that the prevalence of Yq microdeletions was 5.23% (29/554), with 93% (27/29) of the deletions in the AZFc region among 554 infertile men recruited in this study. The results of CMA and multiplex-PCR-based AZFc deletion analysis were generally concordant, but CMA provided more details about location, size and OMIM genes involved in deletion fragments of the AZF region. Of 29 clinically infertile phenotype-related CNVs detected by CMA, nine were pathogenic and the remaining 20 CNVs were OVUS. Except for a 15.69 Mb loss CNV in AZFa + b + c and an 8.25 Mb loss CNV in AZFb + c, others were located in the AZFc region. Based on a combination of the clinical symptoms and loss CNVs, we concluded that the CNV size and the involvement of spermatogenesis critical genes are two important factors that determine the relevance of a CNV in the AZFc region to the presence or absence of a clinically infertile phenotype.


Assuntos
Azoospermia/genética , Variações do Número de Cópias de DNA , Infertilidade Masculina/genética , Oligospermia/genética , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/genética , Adulto , Azoospermia/patologia , Deleção Cromossômica , Cromossomos Humanos Y/genética , Humanos , Infertilidade Masculina/patologia , Masculino , Oligospermia/patologia , Aberrações dos Cromossomos Sexuais , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/patologia
15.
Mol Med Rep ; 21(2): 918-926, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31974623

RESUMO

Isodicentric Y chromosomes are considered one of the most common structural abnormalities of the Y chromosome. Neocentric marker chromosomes, with neocentromeres, have drawn increasing attention in recent years. The present study reported an azoospermic male with a neocentric isochromosome Yp, neo(Yp), and an isodicentric Yq, idic(Yq). The karyotype was analyzed using G­banding, chromosome microarray analysis (CMA), and fluorescence in situ hybridization (FISH) with various detection probes, including sex­determining region on the Y chromosome (SRY) and Y centromeric, applied at the same time. G­banding initially revealed the karyotype 47,X,i(Y)(q10),+mar. CMA indicated the presence of an extra Y chromosome, seemingly equivalent to 47,XYY males. FISH delineated the existence of two centromeres on the idic(Yq). For the marker chromosome, two SRY signals were detected instead of the Y­specific centromere signal, and a visual centromere was observed. This indicated the possible existence of a neocentromere in the marker chromosome, located in the connected region in Yp11.2 band. Finally, the patient's karyotype was established as 47,X,idic(Y)(p11.2), neo(Y)(pter→Yp11.2::Yp11.2→pter). The findings suggested that both idic(Yq) and neo(Yp) could be the main causes of the patient's azoospermia, despite the fact that the partial disomy of Ypter to Yp11.2 did not lead to any major malformations. The present study not only improves the understanding of karyotype/phenotype relationships between neocentric marker Y chromosomes and male infertility, but also supports the hypothesis that the combined application of molecular cytogenetic analysis could aid in reliably confirming breakpoints, origins, and the constitution of the marker chromosomes.


Assuntos
Azoospermia/genética , Cromossomos Humanos Y/genética , Análise Citogenética , Isocromossomos/genética , Adulto , Humanos , Cariotipagem , Masculino
16.
Asian J Androl ; 22(1): 106-111, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31115363

RESUMO

The stromal antigen 3 (STAG3) gene, encoding a meiosis-specific cohesin component, is a strong candidate for causing male infertility, but little is known about this gene so far. We identified STAG3 in patients with nonobstructive azoospermia (NOA) and normozoospermia in the Korean population. The coding regions and their intron boundaries of STAG3 were identified in 120 Korean men with spermatogenic impairments and 245 normal controls by using direct sequencing and haplotype analysis. A total of 30 sequence variations were identified in this study. Of the total, seven were exonic variants, 18 were intronic variants, one was in the 5'-UTR, and four were in the 3'-UTR. Pathogenic variations that directly caused NOA were not identified. However, two variants, c.3669+35C>G (rs1727130) and +198A>T (rs1052482), showed significant differences in the frequency between the patient and control groups (P = 0.021, odds ratio [OR]: 1.79, 95% confidence interval [CI]: 1.098-2.918) and were tightly linked in the linkage disequilibrium (LD) block. When pmir-rs1052482A was cotransfected with miR-3162-5p, there was a substantial decrease in luciferase activity, compared with pmir-rs1052482T. This result suggests that rs1052482 was located within a binding site of miR-3162-5p in the STAG3 3'-UTR, and the minor allele, the rs1052482T polymorphism, might offset inhibition by miR-3162-5p. We are the first to identify a total of 30 single-nucleotide variations (SNVs) of STAG3 gene in the Korean population. We found that two SNVs (rs1727130 and rs1052482) located in the 3'-UTR region may be associated with the NOA phenotype. Our findings contribute to understanding male infertility with spermatogenic impairment.


Assuntos
Azoospermia/genética , Proteínas de Ciclo Celular/genética , Regulação da Expressão Gênica/genética , MicroRNAs/genética , Oligospermia/genética , Espermatogênese/genética , Adulto , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Genótipo , Haplótipos , Humanos , Masculino , Polimorfismo de Nucleotídeo Único , RNA Mensageiro , República da Coreia
17.
Asian J Androl ; 22(1): 100-105, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31134916

RESUMO

Many studies have shown that microRNAs (miRNAs) play vital roles during the spermatogenesis. However, little is known about the altered miRNA profiles of testicular tissues in nonobstructive azoospermia (NOA). Using microarray technology, the miRNA expression profiles of testicular biopsies from patients with NOA and of normal testicular tissues were determined. Bioinformatics analyses were conducted to predict the enriched biological processes and functions of identified miRNAs. The microarray data were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the results of which were then validated with a larger sample size. Correlations between the miRNA expression levels and clinical characteristics were analyzed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic ability of miRNAs for azoospermia. Hierarchical clustering showed that 129 miRNAs were significantly differentially expressed between the NOA and control groups. Bioinformatics analysis indicated that the differentially expressed miRNAs were involved in spermatogenesis, cell cycle, and mitotic prometaphase. In the subsequent qRT-PCR assays, the selected miRNA expression levels were consistent with the microarray results, and similar validated results were obtained with a larger sample size. Some clinical characteristics were significantly associated with the expression of certain miRNAs. In particular, we identified a combination of two miRNAs (miR-10b-3p and miR-34b-5p) that could serve as a predictive biomarker of azoospermia. This study provides altered miRNA profiles of testicular biopsies from NOA patients and examines the roles of miRNAs in spermatogenesis. These profiles may be useful for predicting and diagnosing the presence of testicular sperm in individuals with azoospermia.


Assuntos
Azoospermia/genética , MicroRNAs/metabolismo , Espermatogênese/genética , Testículo/metabolismo , Adulto , Azoospermia/diagnóstico , Biópsia , Análise por Conglomerados , Biologia Computacional , Hormônio Foliculoestimulante/metabolismo , Perfilação da Expressão Gênica , Humanos , Hormônio Luteinizante/metabolismo , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Testosterona/metabolismo , Análise Serial de Tecidos
18.
Cell Prolif ; 53(1): e12726, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31755150

RESUMO

OBJECTIVES: In humans, non-obstructive azoospermia (NOA) is a major cause of male infertility. However, the aetiology of NOA is largely unknown. Previous studies reported that protein CK2ß was abundantly and broadly expressed in spermatogenic cells. Here, we investigate whether protein CK2ß participates in spermatogenesis. MATERIALS AND METHODS: In this study, we separated spermatogenic cells using STA-PUT velocity sedimentation, analysed the expression pattern of protein CK2ß by immunoblotting, specifically deleted Ck2ß gene in early-stage spermatogenic cells by crossing Ck2ßfl mice with Stra8-Cre+ mice and validated the knockout efficiency by quantitative RT-PCR and immunoblotting. The phenotypes of Ck2ßfl/Δ ;SCre+ mice were studied by immunohistochemistry and immunofluorescence. The molecular mechanisms of male germ cell development arrest were elucidated by immunoblotting and TUNEL assay. RESULTS: Ablation of Ck2ß gene triggered excessive germ cell apoptosis, germ cell development arrest, azoospermia and male infertility. Inactivation of Ck2ß gene caused distinctly reduced expression of Ck2α' gene and CK2α' protein. CONCLUSIONS: Ck2ß is a vital gene for germ cell survival and male fertility in mice.


Assuntos
Apoptose/genética , Azoospermia , Caseína Quinase II/deficiência , Células Germinativas , Animais , Azoospermia/enzimologia , Azoospermia/genética , Azoospermia/patologia , Caseína Quinase II/metabolismo , Deleção de Genes , Células Germinativas/enzimologia , Células Germinativas/patologia , Masculino , Camundongos , Camundongos Knockout
19.
Andrologia ; 52(1): e13453, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31762071

RESUMO

miRNAs (MicroRNAs), known as noncoding and important endogenous factors regulating the expression protein-coding genes, are vital regulators in each biological process. Thus, this study aims to explore the key role of four microRNAs in regulating the spermatogenesis. To conduct this experiment, 55 infertile and fertile men provided the study with the sperm and testicular tissue samples. To study the spermatozoa in terms of the morphology, Diff-Quick was applied. Then, quantitative real-time polymerase chain reaction (RT-PCR) was conducted on samples. Our data indicated that in contrast to the miR-15b, significant increasing of miR-383 and miR-122 occurred in both severe oligoasthenoteratozoospermia (SOAT) and moderate oligoasthenoteratozoospermia (MOAT) compared to normal sperm group (N). In addition, it was observed that miR-15b and miR-122 increased in patients with nonobstructive azoospermia (NOA) compared with obstructive azoospermia (OA) group. Expression levels of target genes including P53, CASPASE-9 and CYCLIN D1 underwent principle changes according to miRNAs expression level. Our finding indicated that miRNAs had essential role in the regulation of spermatogenesis, and their expression altering was associated with sperm abnormalities. Thus, microRNAs can be introduced as useful biomarkers to determine male infertility reasons to choose the effective treatment.


Assuntos
Azoospermia/diagnóstico , Redes Reguladoras de Genes , MicroRNAs/metabolismo , Oligospermia/diagnóstico , Espermatogênese/genética , Adulto , Azoospermia/genética , Biomarcadores/análise , Biomarcadores/metabolismo , Caspase 9/genética , Ciclina D1/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Masculino , MicroRNAs/análise , Oligospermia/genética , Espermatozoides/metabolismo , Proteína Supressora de Tumor p53/genética , Adulto Jovem
20.
Andrologia ; 52(2): e13501, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31833082

RESUMO

A patient presenting with azoospermia was referred for genetic evaluation, and upon karyotyping, he was revealed to have two cell lines-mos46,X,ider(Y)(q10)inv(Y)(p11.3q11.1)/45,X. Further cytogenetic studies such as C banding and fluorescence in situ hybridization were performed, which revealed an inversion duplication of a segment of the Y chromosome; hence, the derivative chromosome contained two SRY genes but only one centromere. Y chromosome microdeletion studies were performed in select STS sequences of AZFa, AZFb and AZFc regions and found to be negative for microdeletions. For such a case of infertility, the couple was advised to undergo artificial reproductive techniques with the help of donor spermatozoa.


Assuntos
Azoospermia/genética , Inversão Cromossômica , Isocromossomos , Adulto , Humanos , Cariótipo , Masculino
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