Tissue inhibitor of metalloproteinases (TIMP-1) produced by granulosa and oviduct cells enhances in vitro development of bovine embryo.
Embryogenesis-stimulating activity (ESA) was found in serum-free conditioned media (CM) of bovine cumulus/granulosa cells (BGC) and bovine oviductal epithelial cells (BOEC). The CM of BGC (BGC-CM) contained two molecular species of ESA, one with a low molecular weight (M(r) 30,000) and another with a high molecular weight (M(r) 80,000); but only the activity with low molecular weight was detected in CM of BOEC by gel-permeation chromatography. The smaller ESA (embryogenin-1) in BGC-CM was purified to homogeneity, as a common activity in both CM by a combination of gel-permeation chromatography, ion-exchange chromatography, and reverse-phase HPLC. Embryogenin-1 has a molecular weight of 31,100 (reduced) and has been identified as a bovine tissue inhibitor of metalloproteinase-1 by NH2-terminal amino acid sequence analysis. Western blot analysis, anti-proteinase activities against metalloproteinases, and the nucleotide sequence of cDNA isolated from a lambda gt11 cDNA library of the bovine ovary by a polyclonal antibody against embryogenin-1. These data suggest that the tissue inhibitor of metalloproteinase-1 produced by BGC and BOEC is a major ESA for in vitro development of bovine embryos.
Satoh
T
T
Research Institute for the Functional Peptides, Yamagata, Japan.
Kobayashi
K
K
Yamashita
S
S
Kikuchi
M
M
Sendai
Y
Y
Hoshi
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D017077:Culture Media, Conditioned; D046508:Culture Techniques; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005314:Embryonic and Fetal Development; D005187:Fallopian Tubes / Q000378:metabolism*; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D006107:Granulosa Cells / Q000378:metabolism*; D015151:Immunoblotting; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017385:Sequence Homology; D019714:Tissue Inhibitor of Metalloproteinases
D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D006023:Glycoproteins; D019714:Tissue Inhibitor of Metalloproteinases; D008666:Metalloendopeptidases