Cloning, expression and characterization of mouse spermatid specific thioredoxin-1 gene and protein

Alberto Jiménez, Richard Oko, Jan Åke Gustafsson, Giannis Spyrou, Markku Pelto-Huikko, Antonio Miranda-Vizuete

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

Thioredoxins are proteins that participate in different cellular processes via redox-mediated reactions. For humans, we have recently described two novel members of this family that display a male germ cell specific expression pattern, named spermatid specific thioredoxin (Sptrx-1 and Sptrx-2 respectively). We report here the cloning and characterization of the mouse Sptrx-1 gene and protein, which are similar to those described for the human orthologue. The mouse Sptrx-1 open reading frame encodes for a protein of 462 aa composed of an N-terminal repetitive domain of a 15 residue motif followed by a C-terminal domain typical of thioredoxins. The mouse Sptrx-1 gene sequence is interrupted by only one intron of 525 bp located in the 5′-UTR, and using fluorescence in-situ hybridization we have mapped its chromosomal location to 17E1.2-1.3. Northern blot analysis identified the testis as the only tissue expressing mouse Sptrx-1 mRNA, and by in-situ hybridization we found a strong labelling in the testicular seminiferous tubules, mostly in the round spermatids. Affinity purified antibodies against human Sptrx-1 crossreacted well with the mouse protein confirming its expression in seminiferous tubules at the later stages of spermiogenesis. Recombinant mouse Sptrx-1 displayed protein disulphide reducing activity in an enzymatic assay coupled to NADPH and thioredoxin reductase. The availability of the mouse Sptrx-1 gene sequence is the first step towards the generation of knock-out mice, whose characterization will provide significant information regarding the in-vivo function of Sptrx-1 and its possible implication in several sperm anomalies.

Original languageEnglish (US)
Pages (from-to)710-718
Number of pages9
JournalMolecular Human Reproduction
Volume8
Issue number8
DOIs
StatePublished - 2002

Keywords

  • Fibrous sheath
  • Sperm
  • Testis
  • Thioredoxin

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Genetics
  • Developmental Biology
  • Embryology
  • Cell Biology

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