Plasminogen activator inhibitor-1 gene-deficient mice. I. Generation by homologous recombination and characterization.
Homozygous plasminogen activator inhibitor-1 (PAI-1)-deficient (PAI-1-/-) mice were generated by homologous recombination in D3 embryonic stem cells. Deletion of the genomic sequences encompassing the transcription initiation site and the entire coding regions of murine PAI-1 was demonstrated by Southern blot analysis. A 3.0-kb PAI-1-specific mRNA was identified by Northern blot analysis in liver from PAI-1 wild type (PAI-1+/+) but not from PAI-1-/- mice. Plasma PAI-1 levels, measured 2-4 h after endotoxin (2.0 mg/kg) injection were 63 +/- 2 ng/ml, 30 +/- 10 ng/ml, and undetectable (< 2 ng/ml) in PAI-1+/+, heterozygous (PAI-1+/-) and PAI-1-/- mice, respectively (mean +/- SEM, n = 4-11). PAI-1-specific immunoreactivity was demonstrable in kidneys of PAI-1+/+ but not of PAI-1-/- mice. SDS-gel electrophoresis of plasma incubated with 125I-labeled recombinant human tissue-type plasminogen activator revealed an approximately 115,000-M(r) component with plasma from endotoxin-stimulated (0.5 mg/kg) PAI-1+/+ but not from PAI-1-/- mice, which could be precipitated with a polyclonal anti-PAI-1 antiserum. PAI-1-/- mice were viable, produced similar sizes of litters as PAI-1+/+ mice, and showed no apparent macroscopic or microscopic histological abnormalities.
Carmeliet
P
P
Whitehead Institute for Biomedical Sciences, Cambridge, Massachusetts 02139-4307.
Kieckens
L
L
Schoonjans
L
L
Ream
B
B
van Nuffelen
A
A
Prendergast
G
G
Cole
M
M
Bronson
R
R
Collen
D
D
Mulligan
R C
RC
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002478:Cells, Cultured; D003062:Codon / Q000378:metabolism; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D004622:Embryo, Mammalian; D005260:Female; D017353:Gene Deletion; D015698:Genomic Library; D006720:Homozygote; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis / Q000172:deficiency* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D015183:Restriction Mapping; D013234:Stem Cells / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection
D003062:Codon; D017931:DNA Primers; D018076:DNA, Complementary; D017395:Plasminogen Activator Inhibitor 1; D004247:DNA