The association between Mitochondrial CYB, CO3, ATP6 and ATP8 polymorphisms and male infertility

A genetic predisposition that affects sperm function and performance may account for idiopathic male infertility. Specific conditions of male infertility and abnormal sperm function are linked to genetic changes in the mitochondrial DNA (mtDNA). Certain deficiencies in mitochondrial function may be caused by mutations of the mitochondrial genes MT-CYB, MT-CO3, MT-ATP6 and MT-ATP8. Therefore, the current study objective was to examine how mutations in the MT- CYB, MT-CO3, MT-ATP6 and MT-ATP8 genes affected sperm motility and male infertility. A total of 111 men had their sperm samples collected, of which 67 were subfertile and 44 were fertile. The mitochondrial DNA was isolated and amplified using QIAGEN's QIAamp DNA Mini Kit and REPLI-g Mitochondrial DNA Kit. Following that, PCR and Sanger sequencing were used to find the target sequence in the MT-CYB, MT-CO3, MT-ATP6, and MT-ATP8 genes. In the present research, a total of 49 single nucleotide polymorphisms (SNPs) were identified and genotyped: thirteen SNPs in MT-CYB rs28357685, rs2853508, rs41518645, rs35070048, rs2853507, rs28357376, rs2853506, rs28660155, rs527236194, rs28357373, rs28357369, rs41504845, and rs2854124, twelve SNPs in MT-CO3 gene rs2248727, rs7520428, rs3134801, rs9743, rs28358272, rs2853824, rs2856985, rs2854139, rs41347846, rs28380140, rs3902407, and 28411821, fourteen SNPs in MT-ATP6 rs2001031, rs2000975, rs2298011, rs7520428, rs9645429, rs112660509, rs6650105, rs6594033, rs6594034, rs6594035, rs3020563, rs28358887, rs2096044, and rs9283154, and ten SNPs in MT-ATP8: rs9285835, rs9285836, rs9283154, rs8179289, rs121434446, rs1116906, rs2153588, rs1116905, rs1116907, and rs3020563. The genotypes frequency between fertile and subfertile groups was significantly different for three variants in MT-CYB: rs28357373 (T15629C) (P = 0.0439), rs41504845 (C15833T) ( P = 0.0038), and rs527236194 (T15784C) (P = 0.0005). Additionally, two SNPs demonstrated an association of significance between infertility in men and allele frequency: rs41504845 (C15833T) (P = 0.0147) and rs527236194 (T15784C) (P = 0.0014). Moreover, in MT-CO3 and MT-ATP6, Only the rs7520428 demonstrated a significant difference statistically between subfertile and fertile groups in the genotype and allele frequency test (P < 0.0001 for both). In conclusion, the current study showed that SNPs rs527236194, rs28357373 and rs41504845 in MT-CYB, and rs7520428 in the MT-CO3 and MT-ATP6 were correlated with male infertility. To determine the correct activity of these genes in male infertility, further studies on larger populations are neededDie idiopathische männliche Unfruchtbarkeit kann auf genetische Veranlagungen zurückgeführt werden, die die Leistung und Funktion der Spermien beeinträchtigen. Genetische Veränderungen in der mitochondrialen DNA (mtDNA) wurden mit bestimmten Arten männlicher Unfruchtbarkeit und abnormaler Spermienfunktion in Verbindung gebracht. Mutationen in den Genen MT-CYB, MT-CO3, MTATP6 und MT-ATP8 können zu einigen Mängeln der mitochondrialen Funktion führen. Daher wollten wir in der aktuellen Studie die Wirkung von Mutationen in den MT-CYB, MT-CO3, MT-ATP6 und MT-ATP8 Genen auf die Spermienmotilität und männliche Unfruchtbarkeit untersuchen. Es wurden Samenproben von 111 Männern entnommen, wobei 67 Männer subfertil und 44 fruchtbar waren. QIAamp DNA-Mini Kit und REPLI-g Mitochondrial DNA Kit von QIAGEN wurden verwendet, um die mitochondriale DNA zu isolieren und zu amplifizieren. Gefolgt von PCR und Sanger-Sequenzierung für die Zielsequenz in den MT-CYB, MT-CO3, MT-ATP6, und MT-ATP8 Genen. Insgesamt wurden neunundvierzig Einzelnukleotidpolymorphismen (SNPs) identifiziert und wie folgt genotypisiert: dreizehn SNPs in MTCYB rs2853508, rs28357685, rs41518645, rs2853507, rs28357376, rs35070048, rs2853506, rs28660155, rs527236194, rs28357373, rs28357369, rs41504845, und rs2854124, zwölf SNPs im MTCO3 Gen rs2248727, rs7520428, rs3134801, rs9743, rs28358272, rs2853824, rs2856985, rs2854139, rs41347846, rs28380140, rs3902407 und 28411821, vierzehn SNPs in MT-ATP6 rs2001031, rs2000975, rs2298011, rs7520428, rs9645429, rs112660509, rs6650105, rs6594033, rs6594034, rs6594035, rs3020563, rs28358887, rs2096044, und rs9283154, und zehn SNPs in MT-ATP8: rs9285835, rs9285836, rs9283154, rs8179289, rs121434446, rs1116906, rs2153588, rs1116905, rs1116907, und rs3020563. Bei MT-CYB zeigten drei Varianten einen signifikanten Unterschied in der Häufigkeit der Genotypen zwischen subfertilen und fruchtbaren Gruppen: rs527236194 (T15784C) (P = 0,0005), rs28357373 (T15629C) (P = 0,0439) und rs41504845 (C15833T) (P = 0,0038). Darüber hinaus zeigten zwei SNPs einen signifikanten Zusammenhang zwischen den Allelfrequenzen von rs527236194 (T15784C) (P = 0,0014) und rs41504845 (C15833T) (P = 0,0147) und männlicher Unfruchtbarkeit. Darüber hinaus zeigte nur der rs7520428 bei MT-CO3 und MT-ATP6 einen statistisch signifikanten Unterschied zwischen subfertilen und fruchtbaren Gruppen im Genotyp- und Allel-Frequenztest (P < 0,0001 für beide). Zusammenfassend zeigte die aktuelle Studie, dass die SNPs rs527236194, rs28357373 und rs41504845 bei MT-CYB und rs7520428 bei MT-CO3 und MT-ATP6 mit männlicher Unfruchtbarkeit korreliert waren. Weitere Studien an größeren Populationen sind erforderlich, um die genaue Rolle dieser Gene bei der Entwicklung der männlichen Unfruchtbarkeit aufzudecken

Association between the single nucleotide variants of the mitochondrial cytochrome B gene (MT-CYB) and the male infertility

Background Idiopathic male infertility can be attributed to genetic predispositions that afect sperm performance and function. Genetic alterations in the mitochondrial DNA (mtDNA) have been linked to certain types of male infertility and abnormal sperm function. Mutations in the mitochondrial cytochrome B (MT-CYB) gene might lead to some defciencies in mitochondrial function. Thus, in the current study, we aimed to investigate the efect of mutations in the MT-CYB gene on sperm motility and male infertility. Methods and results Semen specimens were collected from 111 men where 67 men were subfertile and 44 were fertile. QIAamp DNA Mini Kit and REPLI-g Mitochondrial DNA Kit from QIAGEN were used to isolate and amplify the mito chondrial DNA. Followed by PCR and Sanger sequencing for the target sequence in the MT-CYP gene. Sequencing of the MT-CYB gene revealed a total of thirteen single nucleotide polymorphisms (SNPs). Eight SNPs were non-synonymous vari ant (missense variant) including: rs2853508, rs28357685, rs41518645, rs2853507, rs28357376, rs35070048, rs2853506, and rs28660155. While fve SNPs were Synonymous variant: rs527236194, rs28357373, rs28357369, rs41504845, and rs2854124. Among these SNPs, three variants showed a signifcant diference in the frequency of the genotypes between subfertile and fertile groups: rs527236194 (T15784C) (P=0.0005), rs28357373 (T15629C) (P=0.0439), and rs41504845 (C15833T) (P=0.0038). Moreover, two SNPs showed a signifcant association between allelic frequencies of rs527236194 (T15784C) (P=0.0014) and rs41504845 (C15833T) (P=0.0147) and male subfertility. Conclusion The current study showed a signifcant association between the MT-CYB gene polymorphisms and the develop ment of male infertility. In particular, rs527236194, rs28357373 and rs41504845 variants were found to be the most related to the subfertility group. Further studies on larger and other populations are required to reveal the exact role of this gene in the development of male infertility. In addition, functional studies will be helpful to elucidate the molecular impact of the MT-CYP polymorphisms on mitochondrial function

Lack of association between single polymorphic variants of the mitochondrial nicotinamide adenine dinucleotide dehydrogenase 3, and 4L (MT-ND3 and MT-ND4L) and male infertility

Male infertility is a multifactorial condition associated with different genetic abnormalities in at least 15%–30% of cases. The purpose of this study was to identify suspected correlations between infertility and polymorphisms in mitochondrial NADH dehydrogenase subunits 3 and 4L (MT-ND3 and MT-ND4L) in subfertile male spermatozoa. Sanger sequencing of the mitochondrial DNA target genes was performed on 68 subfertile and 44 fertile males. Eight single nucleotide polymorphisms (SNPs) in MT-ND3 (rs2853826, rs28435660, rs193302927, rs28358278, rs41467651, rs3899188, rs28358277 and rs28673954) and seven SNPs in MT-ND4L (rs28358280, rs28358281, rs28358279, rs2853487, rs2853488, rs193302933 and rs28532881) were detected and genotyped. The genotypes and allele frequencies of the study population have shown a lack of statistically significant association between MT-ND3 and MT-ND4L SNPs and male infertility. However, no statistically significant association was found between the asthenozoospermia, oligozoospermia, teratozoospermia, asthenoteratozoospermia, oligoasthenoteratozoospermia and oligoteratozoospermia subgroups of subfertile males. However, rs28358278 genotype of the MT-ND3 gene was reported in the subfertile group but not in the fertile group, which implies a possible role of this SNP in male infertility. In conclusion, the investigated polymorphic variants in the MT-ND3 and MT-ND4L genes did not show any significant association with the occurrence of male infertility. Further studies are required to evaluate these findings. Moreover, the subfertile individuals who exhibit a polymorphism at rs28358278 require further monitoring and evaluation

Mitochondrial nicotinamide adenine dinucleotide hydride dehydrogenase (NADH) subunit 4 (MTND4) polymorphisms and their association with male infertility

Purpose The purpose of the present study was to determine the relationship between infertility and the polymorphisms of mitochondrial NADH dehydrogenase subunit 4 (MTND4) by spermatozoa analysis in fertile and subfertile men. Methods Samples were divided into 68 subfertile men (case group) and 44 fertile men (control group). After semen analysis, samples were purified. The whole genome was extracted using a QIAamp DNA Mini Kit and the mitochondrial DNA was amplified by using the REPLI-g Mitochondrial DNA Kit. Polymerase chain reaction (PCR) was used to amplify the MT-ND4 gene. Then, samples were purified and sequenced using the Sanger method. Results Twenty-five single-nucleotide polymorphisms (SNPs) were identified in the MTND4 gene. The genotype frequencies of the study population showed a statistically significant association between rs2853495 G>A (Gly320Gly) and male infertility (P = 0.0351). Similarly, the allele frequency test showed that rs2853495 G>A (Gly320Gly) and rs869096886 A>G (Leu164Leu) were significantly associated with male infertility (adjusted OR = 2.616, 95% CI = 1.374–4.983, P = 0.002; adjusted OR = 2.237, 95% CI = 1.245–4.017, P = 0.007, respectively). Conclusion In conclusion, our findings suggested that male infertility was correlated with rs2853495 and rs869096886 SNPs in MTND4

A lack of a definite correlation between male sub-fertility and single nucleotide polymorphisms in sperm mitochondrial genes MT-CO3, MT-ATP6 and MT-ATP8

Background An inability of a man to conceive a potentially fertile woman after a year of unprotected intercourse is defned as male infertility. It is reported that 30–40% of males in their reproductive years have abnormalities in sperm production, either qualitatively or quantitatively, or both. However, genetic factors result in up to 15% of male infertility cases. The present study aimed to analyze the possible correlations between sub-fertility and polymorphisms in sperm mitochondrial CO3, ATP6 and ATP8 genes in sub-fertile men. Methods and results For 67 sub-fertile and 44 fertile male samples, Sanger sequencing of selected mitochondrial DNA genes was done. A total of twelve SNPs in the MT-CO3 gene: rs2248727, rs7520428, rs3134801, rs9743, rs28358272, rs2853824, rs2856985, rs2854139, rs41347846, rs28380140, rs3902407, and 28,411,821, fourteen SNPs in the MT-ATP6: rs2001031, rs2000975, rs2298011, rs7520428, rs9645429, rs112660509, rs6650105, rs6594033, rs6594034, rs6594035, rs3020563, rs28358887, rs2096044, and rs9283154, and ten SNPs in the MT-ATP8: rs9285835, rs9285836, rs9283154, rs8179289, rs121434446, rs1116906, rs2153588, rs1116905, rs1116907, and rs3020563 were detected in the case and control groups at diferent nucleotide positions. Only the rs7520428 in the MT-CO3 and MT-ATP6 showed a statistically signifcant diference between sub-fertile and fertile groups in the genotype’s and allele’s frequency test (P<0.0001 for both). Conclusion The results of our study suggest that male sub-fertility is linked with rs7520428 SNP in MT-CO3 and MT-ATP6. The studied polymorphic variations in the MT-ATP8 gene, on the contrary, did not reveal any signifcant association with male sub-fertility