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Matrix degrading proteases in the ovary: expression and function
Umeå University, Faculty of Medicine, Medical Biochemistry and Biophsyics.
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Extracellular matrix degrading proteases from the plasminogen (plg) activator (PA) and the matrix metalloproteinase (MMP) systems have been implicated as important mediators of ovulation and corpus luteum (CL) formation and regression. The aim of this thesis was to investigate the expression and regulation of PAs and MMPs in the ovary and to examine their functional roles for CL formation and function.

The expression of membrane-type MMP-1 (MT1-MMP) and its substrate gelatinase A (MMP-2) mRNAs was studied during pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-induced ovulation in immature rats. These proteases were coordinately regulated so that both were highly expressed in the theca cells of large preovulatory follicles. This suggests that MT1-MMP activates gelatinase A in preovulatory follicles to degrade the follicular wall during ovulation.

In pseudopregnant (psp) rats, MT1-MMP mRNA was expressed in the CL throughout the luteal phase. Tissue inhibitor of metalloproteases type-1 (TIMP-1) mRNA was expressed during CL formation and regression. MMP-2 and collagenase-3 mRNAs were expressed during CL formation and regression, respectively. When the luteal phase was artificially prolonged or shortened, TIMP-1 and collagenase-3 mRNAs were induced only after the serum progesterone levels had decreased, indicating a close association with luteolysis in the rat.

In psp mice, the expression of mRNAs coding for both PAs, seven MMPs, and five protease inhibitors was studied. Most of the studied molecules were coordinately expressed during formation or regression of the CL. However, uPA, MT1-MMP, and TIMP-3 mRNAs were expressed throughout the luteal phase. The role of uPA was examined in psp uPA deficient mice. These mice displayed no abnormalities in luteal function or vascularity. The role of uPA is thus either not essential or can be compensated by other proteases in the absence of uPA.

In order to control the timing of the CL formation, a mouse model for PMSG/hCG-induced CL formation was developed. Five different protocols were evaluated. One of them provided CL that were stable for six days. In that protocol the mice were treated with prolactin (PRL), twice daily from day 2 of CL life onward. The expression of the steroid acute regulatory protein (StAR) mRNA in the psp CL was also characterized to assess its use as a molecular marker for CL development and regression. It was highly expressed in the forming and functional CL and downregulated at a late stage of CL regression.

The functional role of plg and MMPs for CL formation and function was investigated in plg deficient mice treated with the MMP inhibitor galardin (GM6001). Both psp mice and PMSG/hCG +PRL-induced CL formation were used. Several molecular markers for CL development and regression were used to evaluate the health status of the CL. Our data showed that healthy and vascularized CL formed even in plg deficient mice treated with the inhibitor. However, serum progesterone levels were significantly reduced in these mice, an effect that was mainly attributable to the plg deficiency. In conclusion, neither plg nor MMPs, alone or in combination, seem to be essential for the development of a functional CL.

Place, publisher, year, edition, pages
Umeå: Medicinsk biokemi och biofysik , 2004. , 56 p.
Series
Umeå University medical dissertations, ISSN 0346-6612 ; 893
Keyword [en]
Biochemistry, ovary, ovulation, corpus luteum, plasminogen, PA, MMP, rat, mouse
Keyword [sv]
Biokemi
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Medical Biochemistry
Identifiers
URN: urn:nbn:se:umu:diva-280ISBN: 91-7305-659-6 (print)OAI: oai:DiVA.org:umu-280DiVA: diva2:142944
Public defence
2004-06-03, N350, Naturvetarhuset, Umeå Universitet, Umeå, 13:00
Opponent
Supervisors
Available from: 2004-05-12 Created: 2004-05-12Bibliographically approved
List of papers
1. Coordinated and cell-specific regulation of membrane-type matrix metalloproteinase 1 (MT1-MMP) and its substrate matrix metalloproteinase 2 (MMP-2) by physiological signals during follicular development and ovulation
Open this publication in new window or tab >>Coordinated and cell-specific regulation of membrane-type matrix metalloproteinase 1 (MT1-MMP) and its substrate matrix metalloproteinase 2 (MMP-2) by physiological signals during follicular development and ovulation
1998 (English)In: Endocrinology, ISSN 0013-7227, E-ISSN 1945-7170, Vol. 139, no 11, 4735-4738 p.Article in journal (Refereed) Published
Abstract [en]

In the ovary, extensive tissue remodeling is required during both follicular development and the break down of the follicular wall at the time of ovulation. Extracellular proteases such as serine proteases and matrix metalloproteinases (MMPs) are thought to play pivotal roles in these processes. In this paper, we have used in situ hybridization to study the regulation and distribution of mRNA coding for MMP-2 (gelatinase A) and its cell surface activator membrane-type MMP1 (MT1-MMP) during gonadotropin induced ovulation in the rat. In ovaries of untreated immature (23 day old) rats, the levels of MT1-MMP and MMP-2 mRNA were low. MMP-2 mRNA was found in theca-interstitial cells while MT1-MMP mRNA was found in both granulosa and theca-interstitial cells and both messages were induced after stimulation with PMSG. After an ovulatory dose of hCG, the expression of MT1-MMP was dramatically down regulated in the granulosa cell layers of large preovulatory follicles but the expression remained and appeared to be up regulated together with MMP-2 in the theca-interstitial cells surrounding the large preovulatory follicles. The expression kinetics and tissue distribution supports the notion that MT1-MMP may have dual functions in the ovary. Initially MT1-MMP may act as a matrix degrading protease inside the follicle during follicular development and later, just prior to ovulation, as an activator of proMMP-2 in theca-interstitial cells surrounding preovulatory follicles.

Identifiers
urn:nbn:se:umu:diva-4005 (URN)9794486 (PubMedID)
Available from: 2004-05-12 Created: 2004-05-12 Last updated: 2010-09-03Bibliographically approved
2. Distinct expression ofgelatinase A (MMP-2), collagenase-3 (MMP-13), membrane-type MMP 1 (MT1-MMP), and tissue inhibitor of MMPs type 1 (TIMP-1) mediated by physiological signals during formation and regression of the rat corpus luteum
Open this publication in new window or tab >>Distinct expression ofgelatinase A (MMP-2), collagenase-3 (MMP-13), membrane-type MMP 1 (MT1-MMP), and tissue inhibitor of MMPs type 1 (TIMP-1) mediated by physiological signals during formation and regression of the rat corpus luteum
1999 (English)In: Endocrinology, ISSN 0013-7227, E-ISSN 1945-7170, Vol. 140, no 11, 5330-5338 p.Article in journal (Refereed) Published
Abstract [en]

The corpus luteum (CL) is a transient endocrine organ that secretes progesterone to support pregnancy. The CL is formed from an ovulated follicle in a process that involves extensive angiogenesis and tissue remodeling. If fertilization does not occur or implantation is unsuccessful, the CL will undergo regression, which involves extensive tissue degradation. Extracellular proteases, such as serine proteases and matrix metalloproteinases (MMPs), are thought to play important roles in both the formation and regression of the CL. In this study, we have examined the physiological regulation pattern and cellular distribution of messenger RNAs coding for gelatinase A (MMP-2), collagenase-3 (MMP-13), membrane type MMP 1 (MT1-MMP, MMP-14), and the major MMP inhibitor, tissue inhibitor of MMPs type 1 (TIMP-1) in the CL of adult pseudopregnant (psp) rat. Northern blot and in situ hybridization analyses revealed that gelatinase A messenger RNA was mainly expressed during luteal development, indicating that gelatinase A may be associated with the neovascularization and tissue remodeling that takes place during CL formation. Collagenase-3 had a separate expression pattern and was only expressed in the regressing CL, suggesting that this MMP may be related with luteal regression. MT1-MMP that in vitro can activate progelatinase A and procollagenase-3 was constitutively expressed during the formation, function, and regression of the CL and may therefore be involved in the activation of these MMPs. TIMP-1 was induced during both the formation and regression of the CL, suggesting that this inhibitor modulates MMP activity during these processes. To test whether the induction of collagenase-3 and TIMP-1 is coupled with luteal regression, we prolonged the luteal phase by performing hysterectomies, and induced premature luteal regression by treating the pseudopregnant rats with a PGF2alpha analog, cloprostenol. In both treatments, collagenase-3 and TIMP-1 were induced only after the serum level of progesterone had decreased, suggesting that collagenase-3 and TIMP-1 are induced by physiological signals, which initiate functional luteolysis to play a role in tissue degradation during structural luteolysis. In conclusion, our data suggest that gelatinase A, collagenase-3, and MT1-MMP may have separate functions during the CL life span: gelatinase A mainly takes part in CL formation, whereas collagenase-3 mainly takes part in luteal regression; MT1-MMP is constitutively expressed during the CL life span and may therefore serve as an in vivo activator of both gelatinase A and collagenase-3. TIMP-1 is up-regulated both during the formation and regression of the CL and may therefore regulate MMP activity during both processes.

Identifiers
urn:nbn:se:umu:diva-4006 (URN)10537164 (PubMedID)
Available from: 2004-05-12 Created: 2004-05-12 Last updated: 2010-09-03Bibliographically approved
3. Expression pattern and functional studies of matrix degrading proteases and their inhibitors in the mouse corpus luteum
Open this publication in new window or tab >>Expression pattern and functional studies of matrix degrading proteases and their inhibitors in the mouse corpus luteum
2003 (English)In: Molecular and Cellular Endocrinology, ISSN 0303-7207, E-ISSN 1872-8057, Vol. 205, no 1-2, 131-140 p.Article in journal (Refereed) Published
Abstract [en]

The formation of the corpus luteum (CL) is accompanied with angiogenesis and tissue remodeling and its regression involves tissue degradation. Matrix degrading proteases such as plasminogen activators (PAs) and matrix metalloproteinases (MMPs) are thought to play important roles in such controlled proteolytic processes. In this study, in situ hybridization has been used to examine the regulation and expression pattern of mRNAs coding for proteases and protease inhibitors belonging to the PA- and MMP-systems during the life cycle of the CL in an adult pseudopregnant mouse model. Of the nine proteases and five protease inhibitors that were studied, the majority were found to be temporally expressed during the formation and/or the regression of the CL. However, the mRNAs coding for urokinase type PA (uPA), membrane-type 1 MMP (MT1-MMP), and tissue inhibitor of metalloproteinases type-3 (TIMP-3) were constantly expressed in the mouse CL throughout its whole life span. To study the functional role of uPA in the CL, we analyzed luteal formation and function in uPA deficient mice. Our results revealed no significant difference in ovarian weight, serum progesterone levels, and blood vessel density in the functional CL between uPA deficient and wild type control mice. The temporal and spatial expression pattern of proteases and protease inhibitors during the CL life span suggests that members of the PA- and MMP-systems may play important roles in the angiogenesis and tissue remodeling processes during CL formation, as well as in the tissue degradation during luteal regression. However, the absence of reproductive phenotypes in mice lacking uPA and several other matrix degrading proteases indicates that there are redundancies among different matrix degrading proteases or that tissue remodeling in the ovary may involve other additional unique elements.

Keyword
Ovary, Corpus luteum, Plasminogen activators, Matrix metalloproteinase
Identifiers
urn:nbn:se:umu:diva-4007 (URN)10.1016/S0303-7207(03)00147-3 (DOI)12890575 (PubMedID)
Available from: 2004-05-12 Created: 2004-05-12 Last updated: 2011-03-30Bibliographically approved
4. A synchronized gonadotropin-induced corpus luteum model in the mouse
Open this publication in new window or tab >>A synchronized gonadotropin-induced corpus luteum model in the mouse
(English)Article in journal (Refereed) Submitted
Identifiers
urn:nbn:se:umu:diva-4008 (URN)
Available from: 2004-05-12 Created: 2004-05-12 Last updated: 2010-09-03Bibliographically approved
5. Plasminogen is required for normal progesterone production in the mouse
Open this publication in new window or tab >>Plasminogen is required for normal progesterone production in the mouse
(English)Manuscript (Other academic)
Identifiers
urn:nbn:se:umu:diva-4009 (URN)
Available from: 2004-05-12 Created: 2004-05-12 Last updated: 2011-09-12

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