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Molecular characterization of DNAH6 and ATPase6 (Mitochondrial DNA) genes in asthenozoospermia patients in the northern region of India

BMC Urology volume 24, Article number: 180 (2024) Cite this article

Abstract

Background

Male infertility due to spermatogenesis defects affects millions of men worldwide. However, the genetic etiology of the vast majority remains unclear. The present study was undertaken to assess the association of DNAH6 and ATPase6 genes in asthenozoospermia patients in the northern region of India.

Methods

A total of 60 semen samples were collected for the study, of which 30 were from the case group and 30 were from the control group. The semen samples for the case group (asthenozoospermia) and control groups were collected from IVF and Reproductive Biology Centre, Maulana Azad Medical College, New Delhi. Sperm count and motility were classified as per World Health Organization (WHO 2021) protocol. A total genomic DNA was extracted as per the stranded TRIZOL method with little modification.

Results

In-vitro molecular characterizations of DNAH6 and ATPase6 genes in both groups were checked by Polymerase Chain Reaction (PCR). The 675 bp and 375 bp amplicons were amplified using PCR for ATPase6 and DNAH6 genes. Our study results showed a significant (P ≤ 0.05) null deletion of DNAH6 and ATPase6 genes in asthenozoospermia patients as compared to the control. We found the significant null deletion of DNAH6 in case 45.0%, and the control group was 11.7%. However, in the case of APTase6, it was 26.7% and 10.0%, respectively.

Conclusions

Our study concluded that the presence of DHAH6 and ATPase6 genes had a significant impact on male infertility.

Peer Review reports

Background

Infertility is most often seen as a woman’s issue and it is them who bears the brunt of not bearing a child, but according to a recent report by WHO on the status of infertility in the world including in India, of all infertility cases, approximately 50% are due to “male factor” infertility, owing to the reproductive anomalies in the male [1,2,3,4,5,6,7]. Clinically, male infertility is often manifested as poor sperm quality and low motility [8]. Mammalian spermatozoa are formed in the testis through a complex spermatogenesis process and mature in the epididymis by post-translational modification process. Abnormalities of any step-in spermatogenesis or sperm maturation can cause male infertility [6, 9, 10]. Genetic abnormalities have been identified in humans with unidentified asthenozoospermia, including numerical and structural chromosomal abnormalities [11, 12]. Genetic factors involved in male infertility manifest as chromosomal disorders, mitochondrial DNA (mtDNA) mutations, monogenic disorders, multifactorial disorders, and endocrine disorders of genetic origin. Male infertility is often related to chromosome abnormalities or other genetic factors [1, 6, 7, 13].

DNAH6 (Dynein Axonemal Heavy Chain 6) is a protein-coding gene, part of the microtubule-associated motor protein and is a testis specific expressed protein, located in the neck region of the spermatozoa, required for the function of cilia function in spermatozoa [4]. Null deletions of DNAH6 include Mitochondrial DNA Depletion Syndrome 9 and Spermatogenic Failure6. Among its related pathways are respiratory electron transport, ATP synthesis by chemiosmotic coupling, and heat production by uncoupling proteins. Gene ontology annotations related to ATPase6 (Mitochondrial ATPase subunit 6) activity and microtubule motor activity [5].

The aim of the present study was to investigate the molecular characterization of DNAH6 and ATPase6 genes in male infertility in asthenozoospermia patients. As ATPase6 gene actively participates in the formation of ATP (adenosine triphosphate) [1], which is further utilized by the DNAH6 gene for the flagella movement of spermatozoa. Therefore, the molecular investigation of these genes (DNAH6 and ATPase6) in spermatozoa and their role in male fertility may provide an authentic basis for the determination of sperm-embryo interaction, sperm quality and competency in terms of male infertility.

DNAH6 (Dynein Axonemal Heavy Chain 6) is a protein-coding gene [14], part of the microtubule-associated motor protein, which is highly expressed in human and mouse testes [15]. According to Li and co-workers, the DNAH6 gene is a novel candidate gene that is associated with sperm head abnormalities [16]. DNAH6 protein plays an important role in centriole functions like centriole cohesion maintenance and centriole translocation [16,17,18].

Numerous studies concluded that headless spermatozoa were derived from the abnormal implantation of centrioles to the nucleus [19,20,21,22,23]. A null deletion of DNAH6 includes diseases like Mitochondrial DNA Depletion Syndrome 9 and Spermatogenic Failure 6 [24]. Ghieh and colleagues reported that the DNAH6 protein is strongly expressed in the testis and is important for meiosis and ciliary beating [25]. Mutations in DNAH6 have also been linked to primary ciliary dyskinesia and sperm head anomaly, as well as to non-obstructive azoospermia. Apart from nuclear genomes (genes), mitochondria contain their own genomes [26, 27], capable of producing essential components of the respiratory chain which have a profound impact on sperm motility [28]. Mitochondria play an important role in spermatozoa because of the high adenosine triphosphate (ATP) demand of these cells [29,30,31].

Numerous studies have reported that mitochondria contain a thousand distinct proteins involved in many metabolic pathways [32, 33]. Mitochondria contain their own DNA (mtDNA) which codes a few mitochondrial proteins and plays a fundamental role in the production of ATP (energy coin of the cell) through oxidative phosphorylation (OXPHOS) [31, 34]. Mitochondria are exclusively confined in the sperm midpiece and tightly wrapped around the axoneme. Similarly, Prabhu and Sangeetha, 2009 reported that Sperm motility is heavily dependent on the ATP generated by oxidative phosphorylation in the mitochondrial sheath.

Male infertility is characterized by abnormal sperm morphology and/or motility. Good quality ejaculated spermatozoa have hyper-activated motility which requires an adequate supply of energy in the form of ATP that is used by the flagellar dynein ATPase. Oxidative phosphorylation (OXPHOS) is involved in energy production for sperm motility [13, 22, 35]. Research and studies revealed that mitochondrial genes (ATPase, COX, CytB, TSGA, and ND) [2, 36] are playing a crucial role in the formation of mature sperm and flagellar movement after ejaculation [23, 37].

The proposed investigation was based on molecular characterization (null deletion) of DNAH6 and ATPase6 genes in asthenozoospermia patients who attended the IVF and Reproductive Biology Centre at MAMC, New Delhi for infertility treatment. The molecular investigation of DNAH6 and ATPase6 genes in asthenozoospermia patients may help to understand the efficacy of these genes in sperm motility and male infertility.

Methods

All chemicals and media were purchased from Sigma Chemicals Co. (St. Louis, USA) and disposable plasticwares from Tarsons (Pvt. LTD, Martin Burn Business Park, Kolkata, India) unless otherwise mentioned.

Ethical approval

This research was carried out after procuring the necessary approval from the institutional Ethical Committee of the Maulana Azad Medical College, New Delhi, India. Ethical Approval No.170/1,102,020.

Inclusion criterium’s

The study included married couples aged less than 45 years old, who attended IVF clinics at MAMC and with negative viral markers such as Hepatitis B surface antigen and antigen to hepatitis C (HBsAg, Anti-HCV), HIV (human immunodeficiency virus) and syphilis (Treponema pallidum). (Ministry of Health and Family Welfare, ICMR, 2005. (Indian Council of Medical Research).

Semen sample collection

A total of 60 semen samples were taken for investigation (30 from asthenozoospermia patients, i.e., < 39% motility, and 30 from control, i.e., > 39% motility), considering World Health Organization, 2021 protocol for semen analysis. Sample size was calculated by using the n = Z2*P (P-1)/D2 formula. Briefly, semen samples were collected by masturbation in a serial, non-toxic container (Oosafe, USA) and well-labeled with the patient’s name, age, and ID number. After liquefaction of 30 min at 37ºC, the sample was classified as per WHO 2021 guidelines [38]. Semen Analysis was conducting (using a stereomicroscope (Olympus) at 10X in the Makler chamber (SefiMedical instruments, 0.01 seq. mm and 10 μm deep). The sperm count and motility were classified as per WHO, 2021 (Fig. 1).

  • Normal sperm concentration to > 16 million (lower reference value, WHO, 2021).

  • Asthenozoospermia < 39% grade (PR + NR) (lower reference value, WHO, 2021).

Fig. 1
figure 1

Flow diagram showing the overview of the experimental design

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Primer details

DNAH6 and ATPase6 genes were used as reference primers in both case and control groups shown on Table 1 [24, 26]. Amplified PCR products were subjected to electrophoresis in 1.8% agarose gel. The specificity of the size of the amplicons was checked by gel electrophoresis.

Table 1 Primer sequences and information of the DNAH6 and ATPase6 genes
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Genomic DNA isolation, purification, and quantification

Total genomic DNA was extracted from both groups (asthenozoospermia and control). DNA extraction was done by using the TRIZOL method (monophasic solution of phenol and guanidinium isothiocyanate). DNA purification and quantification were done by using a standard protocol with little modification as per Sambrook and Russal’s 2001 protocol [39].

Genomic DNA extraction

Total genomic DNA was extracted from 60 semen samples, each produced from a case and control group by the TRIZOL method. The semen sample was washed with 1X Phosphate buffer saline (PBS) at 12,000 rpm for 10 min. the supernatant was discarded, repeated this step was twice, and the pellet was resuspended in 1 ml of 1X PBS and stored at -20ºC overnight.

Next day, 1 ml of TRIZOL reagent plus 40 µl proteinases-K was added to the suspension and gently mixed by inverting 3–4 times, the mixture incubated at 56 °C for 3 h. 200 µl of chloroform was added and vortexed gently for 30 s. After that, the solution was centrifuged at 12,000 rpm for 10 min. The supernatant was discarded, added 1 ml of absolute ethanol, and centrifuged at 12,000 for 5 min. discard the supernatant, and the pellet was subsequently washed twice with 70% ethanol to dehydrate the DNA at 12,000 rpm for 1 min. The pellet was air-dried to a semi-solid state and then resuspended by adding 50 µl of TE buffer. The concentration of genomic DNA samples was checked by a Nanodrop spectrophotometer (IMPLEN, Bio-Red) at the wavelength of 260/280nm. The integrity of the genomic was checked by 0.8% agarose gel electrophoresis.

Amplification of DNAH6 and ATPase6 gene

The PCR reactions were performed with a final reaction volume of 25 µl having 2X Master mixture (Thermo Scientific), 1 µl of forward and reverse primers (Eurofins, 10pmol), and 1 µl genomic DNA (50ng/µl). The following PCR amplification conditions were used to optimize the reaction accuracy performed in the thermal cycler (ABI, USA): 95 °C for 5 min, 95 °C for 30 s, annealing temperature 56 °C for 1 min, 72 °C for 30 s, and a final extension at 72 °C for 10 min for 35 cycles. Agarose gel electrophoresis was used to detect amplified DNA products. The specificity of amplicon size was confirmed by 375 bp and 675 bp for the DNAH6 and ATPase6 genes on 1.8% agarose gel electrophoresis (Fig. 2). A volume of 10 µl of the amplified product was added with 2 µl of 6X loading dye (Bromophenol blue 0.25% and Sucrose 4.0% w/v) and loaded on 1.8% agarose gel was run at 5 volts/cm for 1 h in 1X TAE buffer (TRIS base, 0.5 M EDTA pH 8.0, Glacial acetic acid). The agarose gel was stained with 0.5 µg/ml. Bands of the amplified product were visualized by exposing the gel to UV light and photographed by a gel documentation system (Bio-Red). The size of the amplicon was measured by comparing it with a standard weight Marker (100 bp).

Fig. 2
figure 2

Amplified PCR product of DNAH6 (375 bp) and ATPase6 (675 bp) genes on 1.8% agarose gel electrophoresis

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Statistical analysis

All parameteric data was used for t-test. Kolmogorov-Smirnov test was used for Normality assessment of the data and SPSS 20 software was used for statistical analysis (Supplementary file S1 and S2 attached).

Availability of data and materials

All data generated or analyzed during this study are included in this article.

Results

Semen sample collection and classification

A total of 60 semen samples were collected from the IVF and reproductive Biology Centre, MAMC, New Delhi, India. Of these, 30 were from asthenozoospermia patients, and the rest 30 were from normal patients (control). From the current investigation (Table 2), we found the average semen volume in the case and control group was 2 ml. However, it can be a variable from person to person. In accordance with the WHO 2021 protocol, the lower reference range is 1.4 ml.

Our data revealed the total sperm count (mill/ml) was 84.20 for the control and 31.53 for the case group. These values are above the minimum reference value of 16 mills/ml (sperm concentration), as per WHO 2021 protocol. We found the mean sperm motility at 60.0% for the control and 27.33% for the case group. Genomic DNA integrity (OD) was 1.7 for both groups, which was further evaluated by agarose gel electrophoresis and polymerase chain reaction to detect the polymorphism of the DNAH6 and ATPase6 genes in both groups.

Table 2 Semen volume, sperm concentration, motility and genomic DNA integrity
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In-vitro molecular characterization of DNAH6 and ATPase6 genes in case and control groups

Amplification of a fragment from the DNAH6 and ATPase6 genes of human semen samples formed 675 bp and 375 bp PCR products, respectively (Fig. 2). The amplification analysis of DNAH6 and ATPase6 genes in both groups revealed the presence of polymorphisms.

Polymerase chain reaction analysis of DNAH6 and ATPase6 gene – control group

Our data revealed a presence of DNAH6 in 23 (38.33%) and absence in 7(11.7%) whereas ATPase6 gene was present in 24(40.0%) and absent in 6(10.0%).

Polymerase chain reaction analysis of DNAH6 and ATPase6 gene – case group

Our data revealed a presence of DNAH6 in 3 (5%) and absence in 27(45%) whereas ATPase6 gene was present in 14(23.3%) and absent in 16(26.7%).

Fig. 3
figure 3

Graph showing the comparative association of DNAH6 (A) and ATPase6 (B) in case and control groups

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Fig. 4
figure 4

Graphs showing the relative association of DNAH6 (A) and ATPase6 (B) genes in case and control groups

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Statistical analysis revealed a higher percentage of ATPase6 and DNAH6 genes detected in both groups (control and case), which was 38 (63.3%) and 26 (43.3%), respectively. However, a lower percentage of both genes, 22 (36.7%), and 34 (56.7%), were detected (Tables 3 and 4; Fig. 3). The difference in the two groups with respect to the two genes was statistically significant. Amplified PCR products were confirmed by agarose gel electrophoresis, which revealed a specific band size of 375 bp and 675 bp for DNAH6 and ATPase6 genes (Fig. 2).

Table 3 DNAH6 in case and control group
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Table 4 ATPase6 in case and control group
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Association of DNAH6 and ATPase6 genes in case and control groups

Data reveals a significantly higher presence of the ATPase6 gene in case 46.7% and control group was 80.0% compared to DNAH6 76.7% and 10.0%, respectively (Tables 5 and 6; Fig. 4).

Table 5 DNAH6 and ATPase6 genes within the control group
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Table 6 DNAH6 and ATPase6 genes within case group
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Discussion

Male infertility is a global medical problem with both physical and psychosocial consequences. asthenozoospermia is particularly prevalent, occurring in ~ 19% of infertile men [40]. Generally, severe asthenozoospermia cases represent a heterogeneous group, and genetically associated asthenozoospermia mainly includes primary ciliary dyskinesia (PCD) and multiple morphological abnormalities of the sperm flagella (MMAF) [31, 40]. Genetic abnormalities including chromosomal disorders, mitochondrial DNA (mtDNA) mutations, monogenic disorders, and multifactorial disorders, have been associated with asthenozoospermia patients [1, 11,12,13]. DNAH6 protein is a part of the microtubule-associated motor protein, which is highly expressed in mammalian testis, especially in human and mice, and required for motile cilia function [13,14,15]. DNAH6 plays a crucial role in centriole function and centriole translocation [18, 21, 24]. ATPase6, a mitochondrial DNA (mtDNA) gene is actively participated in the formation of ATP [1], and utilized by the DNAH6 gene for the flagellar motion of spermatozoa.

Under the present study, the impact of DNAH6 and ATPase6 genes was assessed in asthenozoospermia patients. Our data revealed the direct correlation of null deletion of ATPase6 and DNAH6 genes in asthenozoospermia patients. Our results revealed a significant null deletion of the DNAH6 gene in the case group (asthenozoospermia patients) which was 45.0%. Only 3% of cases showed the presence of the DNAH6 gene. This data supports the hypothesis that the DNAH6 protein contributes to the etiopathology of male infertility. Our study further correlated with several studies where that reported the direct association of the DNAH6 gene with male infertility [16, 22, 41]. Li and co-workers reported that the DNAH6 gene is directly associated with sperm head abnormalities. They also reported that DNAH6 localized in the neck region of normozoospermic, which is directly/indirectly associated with centriolar proteins. However, in the patient that had a complete absence of DNAH6 protein, which may lead to the interaction of DNAH6 and centriole and potentially affect the centriole translocation. The loss of centriole translocation may lead to physical contact between the centriole and nucleus and cause decapitated sperm [16]. However, in accordance with Gershoni et al., a rare mutation in DNAH6 has been shown to be associated with azoospermia patients [15].

In the last few years, mutation in mitochondrial DNA (mtDNA) or ATP-generating genes has been noted to infer changes in sperm motility [7, 33]. The role of mtDNA in male infertility was first correlated in early 1990 when scientists reported reduced sperm motility in individuals having structural defects in mitochondria. Mutations in mtDNA lead to some pathophysiological conditions of mammalian spermatozoa. Moreover, it was noted that multiple mtDNA deletions found frequently in mammalian spermatozoa had reduced low motility [26, 27].

We found the null deletion of ATPase6 (mtDNA) in the asthenozoospermia patients was 26.7% and in the control group, it was 10.0% (Tables 3 and 4). Which was further correlated with a PCR amplicon size of 675 bp (Fig. 2). In a similar study, the deletion of 4977 bp mitochondrial DNA in varicocele patients leads to male infertility [24]. Similarly, other studies also reported the ATPase gene is actively participated in the flagellar movement of spermatozoa and associated with male fertility [24, 41]. Elrahman and associates also reported that the mtDNAs of the ND1, ND2, and ATPase6 genes are positively associated with spermatozoa motility and fertility [6].

Conclusions

On the basis of our findings, we concluded that DNAH6 and ATPase6 (mtDNA) gene integrity and energy maintenance might serve as valuable indicators of sperm quality. Our findings also support the hypothesis that ATPase6 (mitochondrial DNA) and DNAH6 proteins (microtubule-associated motor protein) contribute to the etiopathology of male infertility. Further, the association of DNAH6 and ATPase6 genes with male infertility and pregnancy outcome need to be evaluated using more data-based studies.

Data availability

All data generated or analyzed during this study are included in this article.

Abbreviations

%:

Percentage

ATP:

Adenosine triphosphate

DNAH6:

Dynein Axonemal Heavy Chain 6

ATPase6:

Mitochondrial ATPase subunit 6

DNA:

Deoxyribose nucleic acid

mtDNA:

mitochondrial DNA

OXPHOS:

Oxidative phosphorylation

ETC:

Electron transport chain

PCR:

Polymerase Chain Reaction

bp:

base pairs

WHO:

World Health Organization

HBsAg:

Hepatitis B surface antigen

Anti:

HCV-Antibody to hepatitis C

HIV:

Human immunodeficiency virus

ICMR:

Indian Council of Medical Research

PBS:

Phosphate buffer saline

PCD:

Primary ciliary dyskinesia

MMAF:

Multiple morphological abnormalities of the sperm flagella

References

  1. Holyoake AJ, et al. Association of a novel human mtDNA ATPase6 mutation with immature sperm cells. Andrologia. 2002;31:339–45.

    Article  Google Scholar 

  2. Ferlin A, et al. Male infertility: role of genetic background. Reprod Biomed Online. 2007;14:734–45.

    Article  CAS  PubMed  Google Scholar 

  3. Zegers-Hochschild F, et al. International Committee for Monitoring Assisted Reproductive Technology (ICMART) and the World Health Organization (WHO) revised glossary of ART terminology. Fert Steril. 2009;92:1520–4.

    Article  CAS  Google Scholar 

  4. Li W, et al. Centriole translocation and degeneration during ciliogenesis in Caenorhabditis elegans neurons. EMBO J. 2017;36:2553–66.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Gervasi MG, Visconti PE. Molecular changes and signaling events occurring in spermatozoa during epididymal maturation. Andr. 2017;5:204–18.

    CAS  Google Scholar 

  6. Elrahman ABD, et al. Assessment of correlation between asthenozoospermia and mitochondrial DNA mutations in Egyptian infertile men. J Genet Eng Biotechnol. 2021;19:2–15.

    Google Scholar 

  7. Thomas HIS, et al. Genetic Association in the maintenance of the mitochondrial microenvironment and sperm capacity. Oxid Med Cell Longev. 2021;12:5561395.

    Article  Google Scholar 

  8. Sha YW, et al. TSGA10 is a novel candidate gene associated with acephalic spermatozoa. Clin Genet. 2018;93:776–83.

    Article  CAS  PubMed  Google Scholar 

  9. Cornwall GA. Role of posttranslational protein modifications in epididymal sperm maturation and extracellular quality control. Adv Exp Med Biol. 2014;759:159–80.

    Article  CAS  PubMed  Google Scholar 

  10. Katz DJ, et al. Male infertility-the other side of the equation. Aust Fam Physician. 2017;46:641–6.

    PubMed  Google Scholar 

  11. Chandley AC. Chromosome anomalies and Y chromosome microdeletions as causal factors in male infertility. Hum Reprod. 1998;13:45–50.

    Article  CAS  PubMed  Google Scholar 

  12. Prabhu DK, Sangeetha N. Mitochondrial DNA mutations and male infertility. Indian J Hum Genet. 2009;15:93–7.

    Article  Google Scholar 

  13. Nakada K. Et. Al. Mitochondriarelated male infertility. Pro Natl Acad Sci USA. 2006;103:15148–53.

    Article  CAS  Google Scholar 

  14. Turner RM. Moving to the beat: a review of mammalian sperm motility regulation. Reprod Fertil Deve. 2006;18:25–38.

    Article  Google Scholar 

  15. Gershoni M, et al. A familial study of azoospermic men identifies three novel causative mutations in three new human azoospermia genes. Genet Med. 2017;19:998–1006.

    Article  CAS  PubMed  Google Scholar 

  16. Li L, et al. DNAH6 is a novel candidate gene associated with sperm head anomaly. Andrologia. 2018;200:131–40.

    Google Scholar 

  17. Jones LA, et al. Dynein light intermediate chains maintain spindle bipolarity by functioning in centriole cohesion. J Cell Biol. 2014;207:499–516.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Kumar N, et al. Prevalence of male factor infertility in last ten year at a rural tertiary care of central India: a retrospective analysis. Indian J Obstet Gynecol Res. 2015;2:132–6.

    Article  Google Scholar 

  19. Holstein AF, et al. Dissociated centriole development as a cause of spermatid malformation in man. J Reprod Fertil. 1986;78:719–25.

    Article  CAS  PubMed  Google Scholar 

  20. Baccetti B, et al. Morphogenesis of the decapitated and decaudated sperm defect in two brothers. Gamete Res. 1989;23:181–8.

    Article  CAS  PubMed  Google Scholar 

  21. Ostermeier GC, et al. Reproductive biology delivering spermatozoan RNA to the oocyte. Nature. 2004;13(6988):429.

    Google Scholar 

  22. Kasimanickam V, et al. Association between mRNA abundance of functional sperm function proteins and fertility of Holstein bulls. Theriogenology. 2012;78:2007–19.

    Article  CAS  PubMed  Google Scholar 

  23. Li Y, et al. DNAH6 and its interactions with PCD genes in Heterotaxy and primary ciliary dyskinesia. PLoS Genet. 2016;12:e1005821.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Gashti NG, et al. 4977-bp mitochondrial DNA deletion in infertile patients with varicocele. Andrologia. 2014;46:258–64.

    Article  CAS  PubMed  Google Scholar 

  25. Ghieh F, et al. Genetic defects in human azoospermia. Basic clin Androl. 2019;29:4.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Kao SH, et al. Sperm mitochondrial DNA depletion in men with asthenozoospermia. Fert Steril. 2004;82:66–73.

    Article  CAS  Google Scholar 

  27. Kao SH, et al. Multiple deletions of mitochondrial DNA are associated with the decline of motility and fertility of human spermatozoa. Mol Hum Reprod. 1998;4:657–66.

    Article  CAS  PubMed  Google Scholar 

  28. Vaughan KT, et al. Multiple mouse chromosomal loci for dynein-based motility. Genomics. 1996;36:29–38.

    Article  CAS  PubMed  Google Scholar 

  29. Diez-Sanchez C, et al. Mitochondrial DNA content of human spermatozoa. Biol Reprod. 2003;68:180–5.

    Article  CAS  PubMed  Google Scholar 

  30. Kumar R, et al. Increased lipid peroxidation, low antioxidant levels and nucleotide changes in mitochondrial DNA in idiopathic asthenozoospermic men of Indian origin. Indian J Clin Biochem. 2007;22:355.

    Google Scholar 

  31. Chemes HE, Rawe VY. The making of abnormal spermatozoa: Cellular and molecular mechanisms underlying pathological spermiogenesis. Cell Tissue Res. 2010;341:349–57.

    Article  PubMed  Google Scholar 

  32. Gonczarowska-Jorge H, et al. The proteome of baker’s yeast mitochondria. Mitochondrion. 2017;33:15–21.

    Article  CAS  PubMed  Google Scholar 

  33. Shamsi MB, et al. Genetic and epigenetic factors: role in male infertility. Indian J Urol. 2011;27:110–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Chemes HE. Phenotypes of sperm pathology: genetic and acquired forms in infertile men. J Androl. 2000;21:799–808.

    Article  CAS  PubMed  Google Scholar 

  35. Ford WC. Glycolysis and sperm motility: does a spoonful of sugar help the flagellum go round? Hum Reprod Update. 2006;12:269–74.

    Article  CAS  PubMed  Google Scholar 

  36. Rossella C, et al. New insights into the genetics of spermatogenic failure: a review of the literature. Hum Genet. 2019;138:125–40.

    Article  Google Scholar 

  37. Ambulkar PS, et al. Association of large scale 4977-bp common deletions in sperm mitochondrial DNA with asthenozoospermia and oligoasthenoteratozoospermia. J Hum Reprod Sci. 2016;9:35–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Boitrelle F, Shah R, Saleh R, Henkel R, Kandil H, Chung E, Vogiatzi P, Zini A, Arafa M, Agarwal A. The Sixth Edition of the WHO Manual for Human Semen Analysis: a critical review and SWOT analysis. Life (Basel). 2021;11(12):1368.

    CAS  PubMed  Google Scholar 

  39. Sambrook J, Russell DW, Molecular Cloning. A Laboratory Manual: Cold Spring Harbor Laboratory Press. 2001;3(1).

  40. Curi SM, et al. Asthenozoospermia: analysis of a large population. Arch Androl. 2003;49:343–9.

    Article  CAS  PubMed  Google Scholar 

  41. Tu C, et al. Identification of DNAH6 mutations in infertile men with multiple morphological abnormalities of the sperm flagella. Sci Rep. 2019;1:15864.

    Article  Google Scholar 

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Acknowledgements

The authors thank all people in the Genetic Laboratory, Department of Anatomy), Department of IVF and Reproductive Biology Centre, Maulana Azad Medical College, India for their technical assistance. We would like to thank Dr. Anjali Tempe and all patients for their kind assistance in the collection of semen samples. This study was supported by Maulana Azad Medical College, New Delhi, India.

Funding

This research was supported by “Stg. of Medical Research”, Maulana Azad Medical College, New Delhi, India.

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Authors and Affiliations

  1. Department of Anatomy, Maulana Azad Medical College, New Delhi, 110002, India

    Dinesh Kumar

  2. IVF and Reproductive Biology Centre, Maulana Azad Medical College, New Delhi, 110002, India

    Lalit Mohan Jeena, Anjali Tempe & Renu Tanwar

  3. Viral Research and Diagnostic Laboratory, Department of Microbiology, MGM Medical College, Indore, 452001, India

    Suman Kumar

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  1. Dinesh Kumar
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Contributions

DK, AT, and MLJ conceived and designed the experiments. MLJ performed the experiments and SK worked on statistical analysis of data. AT and RT contributed by providing the semen samples from IVF & reproductive Biology Centre. Finally, DK and MLJ wrote and revised the paper. All authors have read and approved the manuscript.

Corresponding author

Correspondence to Lalit Mohan Jeena.

Ethics declarations

Ethics approval and consent to participate

This research was carried out after procuring the necessary approval from the Institutional Ethical Committee of the Maulana Azad Medical College, New Delhi, India. Ethical Approval No.170/1102020. Informed consent was obtained from the all subjects and/or their legal guardians.

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Not applicable.

Informed consent

All experiments were performed in accordance with relevant guidelines and regulations. All experimental protocols were approved by an institutional committee of Maulana Azad Medical College and informed consent was obtained from all subjects and/or their legal guardian(s).

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The authors declare no competing interests.

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Kumar, D., Jeena, L.M., Tempe, A. et al. Molecular characterization of DNAH6 and ATPase6 (Mitochondrial DNA) genes in asthenozoospermia patients in the northern region of India. BMC Urol 24, 180 (2024). https://doi.org/10.1186/s12894-024-01505-9

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BMC Urology

ISSN: 1471-2490

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