Evaluation of the 5'-flanking regions of murine glutathione peroxidase five and cysteine-rich secretory protein-1 genes for directing transgene expression in mouse epididymis.
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.
Lahti
P P
PP
Department of Physiology, Institute of Biomedicine, University of Turku, FIN-20520 Turku, Finland.
Shariatmadari
R
R
Penttinen
J K
JK
Drevet
J R
JR
Haendler
B
B
Vierula
M
M
Parvinen
M
M
Huhtaniemi
I T
IT
Poutanen
M
M
D000818:Animals; D015152:Blotting, Northern; D004624:Embryo Transfer; D004822:Epididymis / Q000737:chemistry / Q000378:metabolism*; D015870:Gene Expression; D005979:Glutathione Peroxidase / Q000235:genetics*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D008562:Membrane Glycoproteins; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012471:Salivary Proteins and Peptides / Q000235:genetics*; D012670:Seminiferous Epithelium / Q000378:metabolism; D012671:Seminiferous Tubules / Q000378:metabolism; D013091:Spermatogenesis; D013735:Testicular Hormones; D014018:Tissue Distribution; D014162:Transfection
C486321:Crisp1 protein, mouse; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012471:Salivary Proteins and Peptides; D013735:Testicular Hormones; D049452:Green Fluorescent Proteins; D005979:Glutathione Peroxidase; C080439:selenium-independent glutathione peroxidase