RESUMO
Y chromosomal microdeletions at the azoospermia factor locus and chromosome abnormalities have been implicated as the major causes of idiopathic male infertility. A marker chromosome is a structurally abnormal chromosome in which no part can be identified by cytogenetics. In this study, to identify the origin of the marker chromosomes and to perform a genetic diagnosis of patients with azoospermia, two-color fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) techniques were carried out. The marker chromosomes for the two patients with azoospermia originated in the Y chromosome; it was ascertained that the karyotype of both patients was 46,X, ish del(Y)(q11)(DYZ3+, DXZ1-). The combination of two-color FISH and PCR techniques is an important method for the identification of the origin of marker chromosomes. Thus, genetic counseling and a clear genetic diagnosis of patients with azoospermia before intracytoplasmic sperm injection or other clinical managements are important.
Assuntos
Azoospermia/diagnóstico , Aberrações Cromossômicas , Hibridização in Situ Fluorescente , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/diagnóstico , Azoospermia/genética , Azoospermia/patologia , Deleção Cromossômica , Cromossomos Humanos Y/genética , Humanos , Infertilidade Masculina , Cariótipo , Masculino , Reação em Cadeia da Polimerase , Aberrações dos Cromossomos Sexuais , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/genética , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/patologiaRESUMO
The melanocortin 3 receptor (MC3R) gene, which belongs to the rhodopsin-like family A of the G protein-coupled receptor family, plays a crucial role in feed efficiency and energy homeostasis. The aim of this study was to examine associations between bovine MC3R gene polymorphisms and body measurement traits (BMTs) and meat quality traits (MQTs). We identified three synonymous mutations (T429C, T537C, and T663C) in exon 1 of the MC3R gene in Chinese Qinchuan beef cattle (N = 271) by sequencing. D' and r(2) values revealed that these three SNPs were in strong linkage disequilibrium (LD) (r(2) > 0.33); the T429C and T537C SNPs were in complete LD (D' = 1 and r(2) = 1). Association analyses revealed that the SNPs were significantly associated with BMTs and MQTs in Qinchuan cattle. Individuals with the wild homozygotic genotypes g.TTTT and g.TT had significantly higher values of chest depth, heart girth, back fat thickness, intramuscular fat content, and loin muscle area than the mutant heterozygotic genotypes g.TCTC and g.TC. These results suggest that the MC3R gene affects MQTs in Qinchuan cattle, and that it may be a good candidate gene for marker-assisted selection.
Assuntos
Tamanho Corporal/genética , Bovinos/genética , Carne , Polimorfismo de Nucleotídeo Único , Receptor Tipo 3 de Melanocortina/genética , Animais , Bovinos/crescimento & desenvolvimento , Desequilíbrio de LigaçãoRESUMO
Retinitis pigmentosa (RP) is a retinal degenerative disorder that often causes complete blindness. Mutations of more than 50 genes have been identified as associated with RP, including the CACNA1F gene. In a recent study, by employing next-generation sequencing, we identified a novel mutation in the CACNA1F gene. In this study, we used the amplification refractory mutation system (ARMS) and identified a single nucleotide change c.1555C>T in exon 13 of the CACNA1F gene, leading to the substitution of arginine by tryptophan (p.R519W) in a Chinese individual affected by RP. This study actually confirms this novel mutation, and establishes the ARMS technique for the detection of mutations in RP.
Assuntos
Pareamento Incorreto de Bases/genética , Canais de Cálcio Tipo L/genética , Primers do DNA/metabolismo , Mutação de Sentido Incorreto/genética , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , Heterozigoto , Humanos , Dados de Sequência Molecular , Mutação Puntual/genética , Reprodutibilidade dos Testes , Análise de Sequência de DNARESUMO
Non-invasive prenatal diagnosis is used to detect the genetic material of the fetus by isolating the cell-free fetal DNA (cffDNA) from maternal peripheral blood. In order to establish an isolation method for cffDNA from maternal peripheral blood in Chinese women, the cffDNA was acquired with a two-step centrifugation using a QlAamp DNA Blood mini kit. The SRY gene of plasma DNA was amplified by polymerase chain reaction (PCR). Real-time quantitative PCR was used to measure the concentration of cffDNA in maternal peripheral blood in different pregnant women. The results of the SRY gene amplification of plasma DNA from pregnant women was the same as that of the amniocyte DNA. The average concentration of cffDNA in maternal peripheral blood of pregnant women in different gestational stages was 0.98 ng/mL (0.26-1.49 ng/mL), 1.43 ng/mL (0.46- 2.34 ng/mL), and 1.95 ng/mL (0.65-6.81 ng/mL) from early, middle, and late gestational stages, respectively. The mean of cffDNA from total DNA in plasma in different stages of gestation was 22.28% (9.86-27.81%). The lowest concentration of DNA amplified by nested-PCR in our research was 10-4-10-3 ng/µL. The isolation method for cffDNA from maternal peripheral blood was successfully established and further research into its applications will be conducted.
Assuntos
DNA/sangue , Feto , Diagnóstico Pré-Natal/métodos , Fatores de Transcrição SOXB1/sangue , Adulto , Cromossomos Humanos Y/genética , DNA/isolamento & purificação , Feminino , Idade Gestacional , Humanos , Gravidez , Fatores de Transcrição SOXB1/genéticaRESUMO
The aim of this study is to use Y-chromosome gene polymorphism method to investigate regional differences in genetic variation and population evolution history of the Chinese native cattle breeds. Six Y-chromosome short tandem repeat (Y-STR) loci (UMN0929, UMN0108, UMN0920, INRA124, UMN2404, and UMN0103) were analyzed using 1016 healthy and heterogenetic males and 90 females of 9 native cattle breeds (Qinchuan, Jinnan, Zaosheng, Luxi, Nanyang, Jiaxian, Dabieshan, Yanbian, and Menggu) in China. Allele frequency and gene diversity were calculated for the various populations. The results indicated that Y-STRs in the 6 loci have polymorphisms and genetic diversity in Chinese cattle populations. The genetic diversity analysis revealed that the Chinese cattle populations have a close genetic relationship. The analysis of INRA124, UMN2404, and UMN0103 loci revealed the original history of Chinese cattle because of which cattle belonging to Bos taurus or Bos indicus could be determined. Interestingly, a declining zebu introgression was displayed from South to North and from East to West in the Chinese geographical distribution, which implied that cattle population from various regions of China had been subjected to somewhat different evolutionary history. This conclusion supported other evidences such as earlier archaeological, historical research, and blood protein polymorphism analysis.
Assuntos
Cruzamento , Bovinos/genética , Variação Genética , Repetições de Microssatélites/genética , Cromossomo Y/genética , Animais , China , Feminino , Frequência do Gene , Geografia , Haplótipos , Masculino , FilogeniaRESUMO
Silent information regulator 2 (SIRT2), a member of the Sirtuin family of class III nicotinamide adenine dinucleotide-dependent protein deacetylases, plays an important role in senescence, metabolism, and apoptosis. This study was conducted to detect potential polymorphisms of the bovine SIRT2 gene and explore their relationships with meat quality and body measurement traits (BMTs) in Qinchuan cattle. Four single nucleotide polymorphisms (A7445G, C7711T, G17937A, and G20937A) in the fourth intron, fourth exon, ninth exon, and twelfth exon of the SIRT2 gene, respectively, were identified according to the sequencing results of 520 individuals of a Qinchuan cattle population. The genotypic distributions of both A7445G and G20937A were in agreement with the Hardy-Weinberg equilibrium (P < 0.05), whereas the other two mutations were not (0.05 < P < 0.01), based on the X(2) test. Association analysis indicated that the four loci were significantly correlated with several BMTs and meat quality traits. When in combination, the H1H1 (AA-CC-GG-CC) diplotypes showed better BMT and meat quality traits than those by other combinations. Collectively, the results show that SIRT2 is involved in the regulation of the growth and meat quality of cattle, suggesting that the SIRT2 gene may be a candidate gene for marker-assisted selection in the development of future breeding programs for Qinchuan cattle.
Assuntos
Composição Corporal/genética , Bovinos/genética , Polimorfismo de Nucleotídeo Único , Sirtuína 2/genética , Alelos , Animais , Sequência de Bases , Bovinos/crescimento & desenvolvimento , Éxons/genética , Frequência do Gene , Genótipo , Haplótipos , Íntrons/genética , Desequilíbrio de Ligação , Carne/normas , Fenótipo , Análise de Sequência de DNARESUMO
Follicle-stimulating hormone receptor (FSHR), which mediates the functioning of FSH, plays a central role in reproduction. We investigated bovine FSHR gene polymorphisms and analyzed their relationships with pregnancy rates after embryo transfer and with hormone concentrations on the day of embryo transfer. One reported SNP of FSHR, G-278A, located in the 5'-upstream region, was analyzed and three genotypes (GG, GA and AA) were detected in 132 Luxi cattle recipients. Statistical analysis revealed that recipients with the GG genotype had significantly higher estrogen levels on the day of embryo transfer than did GA and AA genotypes. There were no significant differences in pregnancy rates among genotypes, after embryo transfer. We conclude that variation at these loci of the FSHR gene has no significant effect on pregnancy rates in Luxi cattle.
Assuntos
Transferência Embrionária , Estrogênios/sangue , Receptores do FSH/genética , Reprodução/genética , Animais , Bovinos , Feminino , Genótipo , Polimorfismo de Nucleotídeo Único , Gravidez , Taxa de Gravidez , Receptores do FSH/sangueRESUMO
Inhibin is a major regulator of secretion of follicle-stimulating hormone, which is involved in follicular development and regulation of steroidogenesis in females. The objectives of this study were to detect polymorphisms of the bovine inhibin beta-A subunit (INHßA) gene and to evaluate its associations with superovulatory responses in 171 Chinese Holstein cows treated for superovulation. Polymerase chain reaction-restricted fragment length polymorphism revealed a C>T transition determining the StyI polymorphism at position 7639 in intron I of the bovine INHßA gene, and three genotypes (CC, CT, and TT) were detected. The frequencies of the three genotypes showed a tendency for CT > TT > CC, and this polymorphism was in Hardy-Weinberg equilibrium. Statistical analysis revealed no significant differences of least square means for superovulation traits among the three genotypes (P > 0.05). These results demonstrate, for the first time, that the detected loci of the INHßA gene have no significant effects on superovulation performance in Chinese Holstein cows.
Assuntos
Bovinos/genética , Subunidades beta de Inibinas/genética , Polimorfismo Genético , Superovulação/genética , Animais , Estudos de Casos e Controles , Bovinos/fisiologia , FemininoRESUMO
Different molecular aberrations can be discriminated into certain prognostic subgroups in cytogenetically normal acute myeloid leukemia (CN-AML) patients but their impact on allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains controversial and studies from Asian populations are lacking. Forty-two adult non-M3 AML patients receiving allo-HSCT from 2002 to 2009 in southern Taiwan were retrospectively reviewed for survey, 23 (54.7%) of whom were CN-AML. NPM1, FLT3-ITD, and CEBPA were analyzed. After a median follow-up of 104 weeks (range, 8 to 384), patients in the good risk group (harboring either NPM1 or CEBPA mutation without concurrent FLT3-ITD) showed a borderline worse overall survival (OS) compared with the intermediate/poor risk group (P = 0.08). Interestingly, a poorer OS was found in patients with the CEBPA mutation (P = 0.003) but not the NPM1 mutation (P = 0.96). No OS difference was found between patients with or without FLT3-ITD (P = 0.15). In patients receiving allo-HSCT at first remission, there was no significant OS benefit in the good risk group (P = 0.33). In patients receiving allo-HSCT beyond first remission, disease status played a major role (P = 0.006), irrespective of molecular aberrations. Allo-HSCT in good risk patients should be carefully evaluated in Taiwanese, especially in patients with the CEBPA mutation. Conversely, allo-HSCT should be considered in first remission in patients with an intermediate/poor risk, where it may overcome the adverse impact of FLT3-ITD. Disease status remained a main issue in patients receiving allo-HSCT beyond first remission.
Assuntos
Biomarcadores Tumorais/genética , Proteínas Estimuladoras de Ligação a CCAAT/genética , Transplante de Células-Tronco Hematopoéticas , Leucemia Mieloide Aguda/diagnóstico , Proteínas Nucleares/genética , Tirosina Quinase 3 Semelhante a fms/genética , Adulto , Feminino , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/cirurgia , Masculino , Pessoa de Meia-Idade , Mutação , Nucleofosmina , Prognóstico , Resultado do TratamentoRESUMO
Members of the transforming growth factor-ß (TGFß) superfamily are critical regulators of germ cell development that act as extracellular ligands of the signal transduction pathways regulating proliferation, differentiation, apoptosis, and other aspects of cell behavior. Growth differentiation factor 9 (GDF9) is a member of the TGFß superfamily that plays a critical role in ovarian follicular development and ovulation rate in females; however, its role in the testis has not been well elucidated. Therefore, in this study we investigated the effects of GDF9 mutations on the quality of fresh and frozen semen of Holstein bulls. Two reported single nucleotide polymorphisms of GDF9, A485TA and A625C, were analyzed in 129 Holstein bulls. Analysis of variance revealed that the A485T polymorphism had significant effects on the acrosome integrity rate (P < 0.05), whereas the A625T polymorphism was significantly associated with sperm concentration (P < 0.05). In addition, a significant additive effect on sperm concentration was detected for the A485T polymorphism (P < 0.05), whereas the polymorphisms A485TA and A625C had significant dominant effects on acrosome integrity rate and sperm motility in frozen semen, respectively (P < 0.05). This study is the first to show a significant association of GDF9 with sperm quality traits, and the results implied that GDF9 is involved in the initiation or maintenance of spermatogenesis; however, further verification is needed.
Assuntos
Acrossomo/fisiologia , Fator 9 de Diferenciação de Crescimento/genética , Polimorfismo de Nucleotídeo Único , Motilidade dos Espermatozoides/genética , Animais , Animais Endogâmicos , Bovinos , Masculino , Contagem de EspermatozoidesRESUMO
Growth differentiation factor 9 (GDF9) belongs to the transforming growth factor ß superfamily and plays a critical role in ovarian follicular development and ovulation rate. We examined the bovine GDF9 gene polymorphism and analyzed its association with superovulation performance. Based on the sequence of the bovine GDF9 gene, six pairs of primers were designed to detect single nucleotide polymorphisms of two exons and intron 1 of GDF9 using polymerase chain reaction-single-strand conformation polymorphism. Only the products amplified by primer 3-1 displayed polymorphisms. Sequencing revealed two mutations of A485T and A625T in intron 1 of the GDF9 gene in 171 Chinese Holstein cows treated for superovulation. Association analysis showed that these two single nucleotide polymorphisms of A485T and A625T had significant effects on the number of transferable embryos (P < 0.05), and the A625T polymorphism was significantly associated with the total number of ova (P < 0.05). In addition, a significant additive effect on the number of transferable embryos was detected in polymorphisms of A485T (P < 0.05). This study is the first to identify two polymorphisms in bovine GDF9 and describe their correlation with superovulation traits in Chinese Holstein cows.
Assuntos
Bovinos/genética , Fator 9 de Diferenciação de Crescimento/genética , Superovulação/genética , Animais , Feminino , Frequência do Gene , Estudos de Associação Genética , Genótipo , Íntrons , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Chinese native buffaloes have faced the threat of extinction, along with an increase in crossbreeding with domesticated river buffaloes; consequently, conservation of local buffalo genetic resources has become a priority. A Chinese native breed, Jianghan, is often crossed intentionally and unintentionally with imported breeds from India and Pakistan, Murrah, and Nili-Ravi. A total of 128 buffaloes of the breeds Jianghan, Murrah, and Nili-Ravi and their presumed hybrid offspring were genotyped for 10 microsatellite markers. Heterozygosity and Wright's F-statistics were calculated to determine the genetic variation in those populations. The observed average heterozygosities ranged from 0.836 (Murrah) to 0.986 (Jianghan), higher than the expected heterozygosities and all the inbreeding values within the populations were negative. The genetic distances between the presumed hybrid buffaloes and the two imported river type dairy buffalo breeds (Murrah and Nili-Ravi) were lower than with the native Jianghan, indicating strong contributions of the imported breeds to this presumed hybrid buffalo population. This information will be useful for the development of rational breeding for the dairy buffalo industry and for conservation strategies for the Jianghan buffalo.