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Regular Article |
a Departments of Physiology and
b Anatomy, Institute of Biomedicine, University of Turku, FIN-20520 Turku, Finland
c Reproduction & Development Research Group, Blaise Pascal University, CNRS UMR 6547—GEEM, 63177 Aubiere, France
d Research Laboratories of Schering AG, D-13342 Berlin, Germany
ABSTRACT
Based on strong epididymal expression of the mouse glutathione peroxidase 5 (GPX5) and cysteine-rich secretory protein-1 (CRISP-1) genes, we evaluated whether the 5.0-kilobase (kb)-long GPX5 and 3.8-kb-long CRISP-1 gene 5'-flanking regions could be used to target expression of genes of interest into the epididymis in transgenic mice. Of the two candidate promoters investigated, the CRISP-1 promoter-driven enhanced green fluorescent protein (EGFP) reporter gene was highly expressed in the tubular compartment of the testis in all stages of the seminiferous epithelial cycle between pachytene spermatocytes at stage VII to elongated spermatids at step 16. In contrast to CRISP-1, the 5.0-kb 5' region of the mouse GPX5 gene directed EGFP expression to the epididymis. In the various GPX5-EGFP mouse lines, strongest expression of EGFP mRNA was found in the epididymis, but low levels of reporter gene mRNA were detected in several other tissues. Strong EGFP fluorescence was found in the principal cells of the distal caput region of epididymis, and few fluorescent cells were also detected in the cauda region. No EGFP fluorescence was detected in the corpus region or in the other tissues analyzed. Hence, it is evident that the 5.0-kb 5'-flanking region of GPX5 promoter is suitable for directing the expression of structural genes of interest into the caput epididymidis in transgenic mice.
1 This work was founded by grants from the Rockefeller and Ernst-Schering Foundations, the Academy of Finland (project 42145 and 43745), Turku University Foundation, and Turku Graduate School of Biomedical Sciences.
2 Correspondence: Matti Poutanen, Department of Physiology, Institute of Biomedicine, University of Turku, Kiinamyllynkatu 10, FIN-20520 Turku, Finland. FAX: 358 2 2502610; matti.poutanen{at}utu.fi
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