Mutation analysis in patients with total sperm immotility

  • Rute Pereira
  • Jorge Oliveira
  • Luis Ferraz
  • Alberto Barros
  • Rosário Santos
  • Mário Sousa
Genetics

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.

Keywords

Dysplasia of the fibrous sheath (DFS) Genetic diagnosis situs inversus totalis Sperm immotility Whole Exome Sequencing (WES) 

Supplementary material

10815_2015_474_MOESM1_ESM.docx (31 kb)
Supplementary Data 1(DOCX 30 kb)
10815_2015_474_MOESM2_ESM.docx (36 kb)
Supplementary Figure 1(DOCX 36 kb)
10815_2015_474_MOESM3_ESM.docx (148 kb)
Supplementary Figure 2(DOCX 147 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Rute Pereira
    • 1
    • 2
  • Jorge Oliveira
    • 3
    • 4
  • Luis Ferraz
    • 5
  • Alberto Barros
    • 6
    • 7
  • Rosário Santos
    • 3
    • 4
  • Mário Sousa
    • 1
    • 3
  1. 1.Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS)University of Porto (UP)PortoPortugal
  2. 2.Department of Biology, Faculty of SciencesUniversity of PortoPortoPortugal
  3. 3.Multidisciplinary Unit for Biomedical Research-UMIBICBAS-UPPortoPortugal
  4. 4.Molecular Genetics Unit, Centre of Medical Genetics Dr. Jacinto Magalhães (CGMJM)Hospital Centre of Porto (CHP)PortoPortugal
  5. 5.Department of Urology, Hospital Eduardo Santos SilvaHospital Centre of Vila Nova de Gaia (CHVNG)Vila Nova de GaiaPortugal
  6. 6.Centre of Reproductive Genetics Alberto Barros (CGR)PortoPortugal
  7. 7.Department of Genetics, Faculty of MedicineUniversity of PortoPortoPortugal

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