Expression profiling of the developing testis in wild-type and Dazl knockout mice

K Maratou, T Forster, Y Costa, M Taggart, R M Speed, J Ireland, P Teague, D Roy, H J Cooke

Research output: Contribution to journalArticlepeer-review

Abstract

Genetic understanding of malefactor infertility requires knowledge of gene expression patterns associated with normal germ cell differentiation. The mouse is one of the best models of mammalian fertility due to its well-characterized genetics and the existence of many infertile mutants both naturally occurring and experimentally induced. We used cDNA microarrays firstly to investigate normal gene expression in the wild-type (wt) testis and secondly to gain a better insight into the effect of the disruption of the Dazl gene on spermatogenesis. We constructed a cDNA microarray from a subtracted and normalized adult testis library and focused on six developmental time-points during the initial synchronous wave of spermatogenesis. The results suggest that in the wild-type testis, 89.5% of genes on our chip change expression dramatically during the time-course. To identify patterns in the gene-expression data, a k-means clustering algorithm and principal component analysis were used. In the Dazl knockout testes, the majority of genes remain at baseline levels of expression, because absence of Dazl has a severe effect on cell-types present in the testis. Although in the prepubescent Dazl-null mice the final point reached in germ cell development is the leptotene-zygotene stage, the microarray results suggest that lack of Dazl expression has a detectable effect on the mRNA complement of germ cells as early as day 5 when only type A spermatogonia are present.

Original languageEnglish
Pages (from-to)26-54
Number of pages29
JournalMolecular Reproduction and Development
Volume67
Issue number1
DOIs
Publication statusPublished - Jan 2004

Keywords

  • spermatogenesis
  • microarray
  • Dazl
  • testis
  • triple-dye hybridization
  • RNA-BINDING PROTEIN
  • MALE GERM-CELLS
  • GENE-EXPRESSION
  • CDNA LIBRARIES
  • STEM-CELLS
  • MALE-MOUSE
  • SPERMATOGENESIS
  • IDENTIFICATION
  • TRANSCRIPTION
  • SPERMATOGONIA

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