Expression and localisation of extracellular matrix degrading proteases and their inhibitors during the oestrous cycle and after induced luteolysis in the bovine corpus luteum.
The corpus luteum (CL) offers the opportunity to study high proliferative processes during its development and degradation processes during its regression. We examined the mRNA expression of matrix metalloproteases (MMP)-1, MMP-2, MMP-9, MMP-14, MMP-19, tissue inhibitor of MMP (TIMP)-1, TIMP-2, tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), uPA-receptor (uPAR), PA-inhibitors (PAI)-1, PAI-2 in follicles 20 h after GnRH application, CLs during days 1-2, 3-4, 5-7 and 8-12 of the oestrous cycle as well as after induced luteolysis. Cows in the mid-luteal phase were injected with Cloprostenol and the CLs were collected at 0.5, 2, 4, 12, 24, 48 and 64 h after PGF2alpha injection. Real-time RT-PCR determined mRNA expressions. Expression from 20 h after GnRH to day 12: MMP-1, MMP-2, MMP-14 and tPA showed a clear expression, but no regulation. TIMP-1 and uPAR mRNA increased when compared with the follicular phase. TIMP-2, MMP-9, MMP-19 and uPA increased from the follicular phase to days 8-12. PAI-1 and PAI-2 expression increased from days 1-7 and decreased to days 8-12. Induced luteolysis: MMP-1, MMP-2, MMP-9, MMP-14, MMP-19 and TIMP-1 all increased at different time points and intensities, whereas TIMP-2 was constantly decreased from 24 to 64 h. The plasminogen activator system and their inhibitors were up-regulated from 2 to 64 h, tPA was already increased after 0.5 h. Immunohistochemistry for MMP-1, MMP-2, MMP-14: an increased staining for MMP-1 and MMP-14 was seen in large luteal cells beginning 24 h after PGF2alpha application. MMP-2 showed a strong increase in staining in endothelial cells at 48 h.
Kliem
H
H
Physiology Weihenstephan, Technical University Munich, 85354 Freising, Germany.
Welter
H
H
Kraetzl
W D
WD
Steffl
M
M
Meyer
H H D
HH
Schams
D
D
Berisha
B
B
D000818:Animals; D002417:Cattle; D003338:Corpus Luteum / Q000201:enzymology*; D030762:Estrous Cycle / Q000502:physiology; D005109:Extracellular Matrix / Q000201:enzymology*; D005260:Female; D007150:Immunohistochemistry; D003341:Luteolysis / Q000502:physiology*; D020781:Matrix Metalloproteinase 1 / Q000032:analysis; D053511:Matrix Metalloproteinase 14 / Q000032:analysis; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D010447:Peptide Hydrolases / Q000032:analysis*; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis / Q000235:genetics; D017396:Plasminogen Activator Inhibitor 2 / Q000032:analysis / Q000235:genetics; D010960:Plasminogen Activators / Q000032:analysis / Q000235:genetics; D015849:Plasminogen Inactivators / Q000032:analysis / Q000235:genetics; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046509:Tissue Culture Techniques; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000032:analysis / Q000235:genetics; D010959:Tissue Plasminogen Activator / Q000032:analysis / Q000235:genetics
D017395:Plasminogen Activator Inhibitor 1; D017396:Plasminogen Activator Inhibitor 2; D015849:Plasminogen Inactivators; D012333:RNA, Messenger; D019716:Tissue Inhibitor of Metalloproteinase-2; D010447:Peptide Hydrolases; D010960:Plasminogen Activators; D010959:Tissue Plasminogen Activator; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D020781:Matrix Metalloproteinase 1; D053511:Matrix Metalloproteinase 14