Loss of SPEF2 function in mice results in spermatogenesis defects and primary ciliary dyskinesia

Biol Reprod. 2011 Oct;85(4):690-701. doi: 10.1095/biolreprod.111.091132. Epub 2011 Jun 29.

Abstract

Primary ciliary dyskinesia (PCD) results from defects in motile cilia function. Mice homozygous for the mutation big giant head (bgh) have several abnormalities commonly associated with PCD, including hydrocephalus, male infertility, and sinusitis. In the present study, we use a variety of histopathological and cell biological techniques to characterize the bgh phenotype, and we identify the bgh mutation using a positional cloning approach. Histopathological, immunofluorescence, and electron microscopic analyses demonstrate that the male infertility results from shortened flagella and disorganized axonemal and accessory structures in elongating spermatids and mature sperm. In addition, there is a reduced number of elongating spermatids during spermatogenesis and mature sperm in the epididymis. Histological analyses show that the hydrocephalus is characterized by severe dilatation of the lateral ventricles and that bgh sinuses have an accumulation of mucus infiltrated by neutrophils. In contrast to the sperm phenotype, electron microscopy demonstrates that mutant respiratory epithelial cilia are ultrastructurally normal, but video microscopic analysis shows that their beat frequency is lower than that of wild-type cilia. Through a positional cloning approach, we identified two sequence variants in the gene encoding sperm flagellar protein 2 (SPEF2), which has been postulated to play an important role in spermatogenesis and flagellar assembly. A causative nonsense mutation was validated by Western blot analysis, strongly suggesting that the bgh phenotype results from the loss of SPEF2 function. Taken together, the data in this study demonstrate that SPEF2 is required for cilia function and identify a new genetic cause of PCD in mice.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Ciliary Motility Disorders / pathology
  • Ciliary Motility Disorders / physiopathology*
  • Epididymis / metabolism
  • Epididymis / ultrastructure
  • Hydrocephalus / physiopathology
  • Infertility, Male / metabolism
  • Infertility, Male / pathology*
  • Male
  • Maxillary Sinusitis / immunology
  • Maxillary Sinusitis / pathology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Mutant Strains
  • Molecular Sequence Data
  • Neutrophil Infiltration
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / physiology*
  • Respiratory Mucosa / physiopathology
  • Respiratory Mucosa / ultrastructure
  • Sperm Tail / metabolism
  • Sperm Tail / ultrastructure
  • Spermatids / metabolism
  • Spermatids / ultrastructure
  • Spermatogenesis
  • Spermatozoa / metabolism
  • Spermatozoa / ultrastructure
  • Trachea / physiopathology
  • Trachea / ultrastructure

Substances

  • Proteins
  • SPEF2 protein, mouse

Associated data

  • GENBANK/HQ856050