title: Mutation analysis in patients with total sperm immotility.
abstract: OBJECTIVE
  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.
authors: 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.
authors: Rute
authors: R
authors: Pereira
authors: Jorge
authors: J
authors: Oliveira
authors: Luis
authors: L
authors: Ferraz
authors: Alberto
authors: A
authors: Barros
authors: Rosário
authors: R
authors: Santos
authors: Mário
authors: M
authors: Sousa
mesh_terms: D004252:DNA Mutational Analysis; D059472:Exome; D005819:Genetic Markers; D006801:Humans; D007248:Infertility, Male / Q000235:genetics* / Q000473:pathology; D008297:Male; D009154:Mutation; D013081:Sperm Motility / Q000235:genetics*
chemical_list: D005819:Genetic Markers