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Mutation analysis in patients with total sperm immotility

  • Genetics
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Abstract

Purpose

Perform the genetic characterization of five patients with total sperm immotility using Sanger sequencing and Whole Exome Sequencing (WES), in order to increase the knowledge on the genetics of sperm immotility and, ultimately, allow the identification of potential genetic markers for infertility.

Methods

Prospective study at a University Medical school. We analysed five men with total sperm immotility, four with dysplasia of the fibrous sheath (DFS), associated with disruption of several axonemal structures, and one patient with situs inversus totalis, which showed absence of dynein arms (DA) and nexin bridges. We screened 7 genes by Sanger sequencing, involved in sperm motility and associated to ultrastructural defects found in these patients (CCDC39, CCDC40, DNAH5, DNAI1, RSPH1, AKAP3 and AKAP4). Additionally, we performed WES analysis in the patient with situs inversus.

Results

We identified nine new DNA sequence variants by WES. Two of these variants were considered particularly relevant: a homozygous missense change in CCDC103 gene (c.104G > C, p.R35P) probably related with absence of dynein arms; the other in the INSL6 gene (c.262_263delCC) is thought to be also involved in sperm immotility.

Conclusions

Our work suggests that WES is an effective strategy, especially as compared with conventional sequencing, to study highly heterogenic genetic diseases, such as sperm immotility. For future work we expect to expand the analysis of WES to the other four patients and complement findings with expression analysis or functional studies to determine the impact of the novel variants.

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Acknowledgements

We would like to acknowledge: Helena Oliveira, MSc, ESHRE Senior clinical embryologist; Ilda Pires, MSc, ESHRE Senior clinical embryologist and Madalena Cabral, BSc, Embryologist, for semen processing at CHVNG; Ana Gonçalves, BSc, and Cláudia Osório, BSc, for semen processing at CGR; Elsa Oliveira, 1st Class Technical Specialist of Pathology, Cytology and Thanatology in the Area of Diagnosis and Therapy and Ângela Alves, Technical assistant teaching and research, for semen processing for electron microscopy at ICBAS-UP.

We also would like to acknowledge Conceição Egas, PhD and Hugo Froufe, MSc (GenoInseq) for performing exome sequencing and assisting the initial variant filtering/analysis.

Funding

This work was financed by the Institutions of the authors and in part by UMIB, which is funded by National Funds through FCT-Foundation for Science and Technology, under the Pest-OE/SAU/UI0215/2014.

Disclosure statement

The authors have nothing to declare.

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Authors and Affiliations

  1. Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal

    Rute Pereira & Mário Sousa

  2. Department of Biology, Faculty of Sciences, University of Porto, Rua do Campo Alegre 1021/1055, 4169-007, Porto, Portugal

    Rute Pereira

  3. Multidisciplinary Unit for Biomedical Research-UMIB, ICBAS-UP, Porto, Portugal

    Jorge Oliveira, Rosário Santos & Mário Sousa

  4. Molecular Genetics Unit, Centre of Medical Genetics Dr. Jacinto Magalhães (CGMJM), Hospital Centre of Porto (CHP), Praça Pedro Nunes, 88, 4099-028, Porto, Portugal

    Jorge Oliveira & Rosário Santos

  5. Department of Urology, Hospital Eduardo Santos Silva, Hospital Centre of Vila Nova de Gaia (CHVNG), Espinho, E.P.E., Rua Conceição Fernandes, 4434-502, Vila Nova de Gaia, Portugal

    Luis Ferraz

  6. Centre of Reproductive Genetics Alberto Barros (CGR), Av. do Bessa, 240, 1º Dto. Frente, 4100-012, Porto, Portugal

    Alberto Barros

  7. Department of Genetics, Faculty of Medicine, University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319, Porto, Portugal

    Alberto Barros

Authors
  1. Rute Pereira
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  2. Jorge Oliveira
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  3. Luis Ferraz
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  4. Alberto Barros
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  5. Rosário Santos
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  6. Mário Sousa
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Corresponding author

Correspondence to Mário Sousa.

Additional information

Capsule We studied four patients with fibrous sheath dysplasia and one with situs inversus totalis, all with total sperm immotility. Nine new DNA sequence variants were identified. Whole Exome Sequencing (WES) revealed to be the most efficient procedure for the genetic analysis.

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Pereira, R., Oliveira, J., Ferraz, L. et al. Mutation analysis in patients with total sperm immotility. J Assist Reprod Genet 32, 893–902 (2015). https://doi.org/10.1007/s10815-015-0474-6

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  • DOI: https://doi.org/10.1007/s10815-015-0474-6

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