Type IV collagenase is a metalloproteinase that specifically cleaves type IV collagen, the major structural component of
basement membranes. The metastatic potential of tumor cells has been found to correlate with the activity of this enzyme.
Type IV collagenase, 72-kD, is officially designated matrix metalloproteinase-2 (MMP2). It is also known as gelatinase,
72-kD.
NCBI Summary:
This gene is a member of the matrix metalloproteinase (MMP) gene family, that are zinc-dependent enzymes capable of cleaving components of the extracellular matrix and molecules involved in signal transduction. The protein encoded by this gene is a gelatinase A, type IV collagenase, that contains three fibronectin type II repeats in its catalytic site that allow binding of denatured type IV and V collagen and elastin. Unlike most MMP family members, activation of this protein can occur on the cell membrane. This enzyme can be activated extracellularly by proteases, or, intracellulary by its S-glutathiolation with no requirement for proteolytical removal of the pro-domain. This protein is thought to be involved in multiple pathways including roles in the nervous system, endometrial menstrual breakdown, regulation of vascularization, and metastasis. Mutations in this gene have been associated with Winchester syndrome and Nodulosis-Arthropathy-Osteolysis (NAO) syndrome. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Oct 2014]
General function
Enzyme, Hydrolase, Peptidase/Protease
Comment
Characterization of Matrix Metalloproteinase-2 and -9 and their inhibitors in equine granulosa cells in vivo and in vitro. Sessions DR et al. Matrix metalloproteinases (MMP) and tissue inhibitors of MMP (TIMP) regulate tissue remodeling events necessary for ovulation. Thus, changes in MMP and TIMP expression and protein enzyme activity were examined in vivo and in vitro during follicular development and atresia in the horse. Equine granulosa cells and follicular fluid from medium (15 to 29 mm) healthy and atretic follicles and from large (>30 mm) healthy and preovulatory follicles were collected by transvaginal aspiration. The cells were either snap frozen (in vivo study) or cultured for 48 h (in vitro study) to determine gene expression and protein enzyme activity of MMP-2 and -9 and TIMP-1 and -2. Concentrations of progesterone and estradiol were determined by RIA in follicular fluid and conditioned media and were used along with follicle dynamics to classify follicles. In vivo, expression of MMP-2 and TIMP-2 was increased (P < 0.05) in large preovulatory follicles, while TIMP-1 was decreased. The ratio of MMP-2:TIMP-2 expression was decreased (P < 0.05) in medium healthy and large preovulatory follicles, while MMP-9:TIMP-1 ratio was increased only in large preovulatory compared to large healthy follicles. Estradiol was greatest (P < 0.05) in fluid of large healthy and large preovulatory follicles. However, medium atretic follicles were associated with the lowest estradiol concentrations, both in vivo and in vitro. Progesterone concentrations were greatest (P < 0.05) in large preovulatory follicles both in vivo and in vitro. In healthy follicles in vivo, the diameter correlated to estradiol concentration, estradiol:progesterone ratio, MMP-9 and TIMP-1 expression, and MMP-2 and -9 protein activity. In contrast to in vivo studies, the ratio of MMP-9:TIMP-1 expression was increased (P < 0.05) in medium healthy follicles; TIMP-2 expression decreased in large preovulatory follicles in vitro. In addition, MMP-9 protein activity was decreased (P < 0.05) in the media samples of cells from large healthy compared to medium healthy follicles. These results indicate changes in MMP-2 and -9 activities may be essential to the tissue reorganization necessary for ovulation in the equine ovary.
A Follicle Rupture Assay Reveals an Essential Role for Follicular Adrenergic Signaling in Drosophila Ovulation. Deady LD et al. (2015) Ovulation is essential for the propagation of the species and involves a proteolytic degradation of the follicle wall for the release of the fertilizable oocyte. However, the precise mechanisms for regulating these proteolytic events are largely unknown. Work from our lab and others have shown that there are several parallels between Drosophila and mammalian ovulation at both the cellular and molecular levels. During ovulation in Drosophila, posterior follicle cells surrounding a mature oocyte are selectively degraded and the residual follicle cells remain in the ovary to form a corpus luteum after follicle rupture. Like in mammals, this rupturing process also depends on matrix metalloproteinase 2 (Mmp2) activity localized at the posterior end of mature follicles, where oocytes exit. In the present study, we show that Mmp2 activity is regulated by the octopaminergic signaling in mature follicle cells. Exogenous octopamine (OA; equivalent to norepinephrine, NE) is sufficient to induce follicle rupture when isolated mature follicles are cultured ex vivo, in the absence of the oviduct or ovarian muscle sheath. Knocking down the alpha-like adrenergic receptor Oamb (Octoampine receptor in mushroom bodies) in mature follicle cells prevents OA-induced follicle rupture ex vivo and ovulation in vivo. We also show that follicular OA-Oamb signaling induces Mmp2 enzymatic activation but not Mmp2 protein expression, likely via intracellular Ca2+ as the second messenger. Our work develops a novel ex vivo follicle rupture assay and demonstrates the role for follicular adrenergic signaling in Mmp2 activation and ovulation in Drosophila, which is likely conserved in other species.//////////////////
Matrix metalloproteinase 2 level in human follicular fluid is a reliable marker of human oocyte maturation in in vitro fertilization and intracytoplasmic sperm injection cycles. Yang WJ et al. (2015) To determine whether matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMP-1 and TIMP-2) in human follicular fluid, have any relationships with oocyte maturation in vivo and subsequent fertilization during in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) cycles. The follicular fluids were obtained from 150 female patients undergoing IVF/ICSI cycles and a total of 1504 oocytes were retrieved for analysis. MMP-2 and MMP-9 activities were measured using zymography assay. TIMP-1 and TIMP-2 concentrations were quantitatively assessed using enzyme-linked immunosorbent assay (ELISA). Human follicular fluid MMP-2 level was significantly associated with the rate of maturity of oocytes (P < 0.001). Furthermore, the MMP-2 was significantly associated with the higher fertilization rate (P < 0.01). There was no significant correlation between follicular MMP-9 and the maturation rate of oocytes. The TIMP-1 and TIMP-2 also showed no correlation with the oocyte maturation rate. The level of gelatinase MMP-2 in human follicular fluid might be a reliable marker of mature oocytes during IVF/ICSI cycles. Furthermore, the MMP-2 expression has a strong association with higher fertilization rate. Further studies are needed to support this theory.//////////////////
LH-Induced Steroidogenesis in the Mouse Ovary, but Not Testis, Requires Matrix Metalloproteinase 2 and 9-Mediated Cleavage of Upregulated EGF Receptor Ligands. Light A et al. (2015) Oocyte maturation and cumulus cell expansion depend on LH-mediated upregulation of membrane bound EGF-like ligands, including amphiregulin, epiregulin, and betacellulin. These ligands then trans-activate the EGF receptor (EGFR) after release by matrix metalloproteinases (MMPs). However, direct measurement of released EGF-like ligands or MMPs from granulosa cells has not been formally evaluated, nor has direct identification of responsible MMPs. Here we address these issues by analyzing LH-induced steroidogenesis, which is also MMP- and EGFR-dependent, in freshly isolated mouse primary granulosa cells. We demonstrate a correlation between amphiregulin and epiregulin mRNA induction and steroid production in LH-treated granulosa cells as well as in ovaries of hCG-treated mice. In contrast, LH does not alter Mmp1, Mmp2, Mmp3, Mmp8, Mmp9, or Adam17 mRNA expression. We demonstrate that, in primary mouse granulosa cells, LH triggers release of soluble amphiregulin that correlates with steroid production, both of which are blocked by MMP2/9 inhibition, confirming that MMP2/9 likely regulates LH-induced amphiregulin release and downstream processes. Notably, LH does not alter secretion of MMP2/9 from primary granulosa cells, nor does it modulate MMP activity. These findings indicate that, in the ovary, LH dictates EGFR-mediated processes not by regulating MMPs, but instead by increasing EGF-like ligand availability. In contrast, LH stimulation of primary mouse Leydig cells does not induce EGF-like ligand expression or require MMP2/9 for steroidogenesis, confirming marked differences in LH receptor-induced processes in the testes. Our results suggest that MMP inhibition may be a means of attenuating excess ovarian steroid production in diseases like polycystic ovary syndrome.//////////////////
The preovulatory surge of gonadotropins activates a cascade of proteolytic enzymes resulting in the rupture of the follicular
wall and the release of a fertilizable ovum during ovulation. Several studies revealed a preovulatory increase in ovarian collagenolytic activity in vivo and an increase in activatable collagenase in vitro.
In view of the complicated control of mammalian collagenase synthesis and activity by local inhibitors and activators, Reich et al examined the expression of ovarian interstitial and type IV collagenases and tissue inhibitor of metalloproteinase (TIMP) mRNA after an ovulatory stimulus. Ovarian mRNA was isolated from immature PMSG-treated rats 3, 6, and 9 h after hCG stimulation. Northern blot analyses revealed a mRNA of 1.7 kilobases (kb) hybridizing with the human interstitial collagenase cDNA probe. The levels of this mRNA showed a 25-fold increase between 3-6 h after hCG stimulation. The human cDNA probe of collagenase IV hybridized with a mRNA of 3.1 kb, which showed only a 4-fold increase 9 h after hCG treatment. The interstitial collagenase mRNA was expressed in both granulosa cells of preovulatory follicles and the residual ovarian tissue, whereas the expression of collagenase IV mRNA was limited to the residual tissue.
Role of matrix metalloproteinase 2 in the ovulatory folliculo-luteal transition of ewes. Gottsch ML et al. Tissue dissolution and remodelling are associated with the processes of rupture of the ovulatory follicle and formation of the corpus luteum. Matrix metalloproteinase 2 (MMP-2) belongs to a family of endopeptidases that cleave extracellular proteins; its primary substrate is the lattice network of basement membranes that support epithelial cells and endothelium. The aim of this study was to ascertain a putative regulatory role of MMP-2 relevant to the folliculo-luteal transformation in ewes. Luteal regression and the preovulatory surge of gonadotrophins were synchronized by administration of PGF(2alpha) and GnRH on days 14.0 and 15.5 of the oestrous cycle, respectively. Dominant antral follicles present during pro-oestrus consistently ovulate approximately 24 h after GnRH administration. Normal IgG or a bioactivity-neutralizing MMP-2 monoclonal antibody was injected into the antral cavity of preovulatory follicles at 8 h after GnRH administration. Jugular blood samples were obtained for serum progesterone analysis and ovaries were removed for light microscopic morphometry on day 8. A definitive ovulation stigma was evident in control ewes. The antra of ruptured follicles had largely been supplanted with luteal tissue. In contrast, the ovarian surface contiguous with follicles injected with anti-MMP-2 was smooth and undisturbed, which is indicative of a failure of ovulation. Luteinized unruptured follicles were filled with (entrapped) fluid. Corpora lutea of control animals contained numerous connective tissue projections that provided a framework for cellular migration and angiogenesis. Luteal tissues that surrounded the cavity of antibody-treated follicles lacked trabeculae and were deficient in blood vessels. Systemic venous progesterone concentrations were lower in ewes with a luteinized unruptured follicle compared with those with a corpus luteum. It is proposed that MMP-2 is a mediator of ovulation and luteal development.
Basal membrane remodeling during follicle histogenesis in the rat ovary: contribution of proteinases of the MMP and PA families Mazaud S, et al .
In mammalian females, follicular units arise from the fragmentation of ovigerous cords, which spread over the first three postnatal days in the rat. The mechanisms underlying such a process of epithelial remodeling involve a specific balance between basal membrane (BM) deposition and degradation that has as yet not been precisely described. We have investigated the contribution of proteases in BM remodeling by localization of transcripts, protein, or enzymatic activity. In addition, we have analyzed BM deposition at the ultrastructural level and by immunofluorescence detection of BM components. At birth, when fragmentation occurred, epithelial cells displayed an upregulation of membrane type 1-matrix metalloproteinase (MT1-MMP) and urokinase-type plasminogen activator (uPA), as well as laminin alpha1 mRNAs. Although MMP2 expression was restricted to mesenchymal cells throughout development, in situ zymography showed that gelatinase-MMP2 activity colocalized with BM deposition inside deepening clefts in the areas of ovigerous cord fragmentation. In the days following birth, gelatin and plasminogen-casein zymography showed an increased enzymatic activity of MMP2 and uPA, respectively. In organotypic cultures of 21-day postconception ovaries, serine protease inhibitors like aprotinin could efficiently block follicle histogenesis. In addition, our results show that the well described and great wave of oocyte attrition characteristic of the days following birth closely correlates with BM remodeling. Altogether, our data show that during follicle histogenesis, ovigerous cord fragmentation results from an acute BM component deposition in deepening clefts and that BM homeostasy involves proteinases of the MMP2/MT1-MMP/TIMP3 and plasminogen/uPA families.
Gelatinase A and membrane-type matrix metalloproteinases 1 and 2 are responsible for follicle rupture during ovulation in the medaka Ogiwara K, et al .
Identification of the hydrolytic enzymes involved in follicle rupture during vertebrate ovulation remains a central challenge for research in reproductive biology. Here, we report a previously uncharacterized approach to this problem by using an in vitro ovulation system in the medaka, Oryzias latipes, which is a small freshwater teleost. We found that follicle rupture in the medaka ovary involves the cooperation of at least three matrix metalloproteinases (MMPs), together with the tissue inhibitor of metalloproteinase-2b protein. We determined the discrete roles of each of these proteins during follicle rupture. Our results indicated that gelatinase A induces the hydrolysis of type IV collagen constituting the basement membrane, membrane-type 2 MMP degrades type I collagen present in the theca cell layer, and MT1-MMP and the tissue inhibitor of metalloproteinase-2b are involved in the production and regulation of gelatinase A. These findings will help clarify the mechanism of follicle wall degradation during ovulation in mammalian species.
Expression regulated by
LH, Steroids, Growth Factors/ cytokines
Comment
Hwang JJ et al examined a possible mechanism for the effect of relaxin on ovulation, granulosa cells and theca-interstitial cells were obtained from ovaries of eCG-primed immature rats. In the granulosa cell culture, relaxin increased the secretion of two major gelatinases of about 92 and 63 kDa in a dose-and time-dependent manner within 24 h of treatment. In
the theca-interstitial cell culture, relaxin induced dose- and time-dependent increases in the secretion of two other major
gelatinases of about 76 and 71 kDa. These gelatinases were characterized as metalloproteinases but not serine/cysteine
proteinases. Furthermore, an immunoblot study demonstrated that relaxin stimulated the secretion of a 72-kDa type IV
collagenase-like substance from cultured theca-interstitial cells but not from granulosa cells.
Puistola U, et al. reported the regulation of 72-kd type IV collagenase-matrix metalloproteinase-2 by estradiol and gonadotropin-releasing hormone agonist in human granulosa-lutein cells. Estradiol increases the matrix metalloproteinase-2 enzyme amount in granulosa-lutein cells; the effect may be antagonized by GnRH-a.
Goto T, et al. reported that gonadotropin-releasing hormone agonist has the ability to induce increased matrix metalloproteinase (MMP)-2 and membrane type 1-MMP expression in corpora lutea, and structural luteolysis in rats.
Ovarian localization
Granulosa, Theca, Luteal cells, Follicular Fluid
Comment
Matrix metalloproteinases and their inhibitors in human cumulus and granulosa cells as biomarkers for oocyte quality estimation. Luddi A et al. (2018) To study the molecular profile of metalloproteinases and their tissue inhibitors in granulosa and cumulus cells in a subset of fertile and infertile women. Molecular study with granulosa and cumulus cells. University hospital. Forty-four women undergoing assisted reproductive techniques for female infertility factor, with partners having a normal spermiogram and 15 normally fertile women with male partner affected by severe oligoasthenoteratozoospermia or nonobstructive azoospermia. In vitro fertilization. We investigated gene expression level of metalloproteinases (MMP2, MMP9, MMP11) and their tissue inhibitors (TIMP1, TIMP2) by means of quantitative reverse-transcription polymerase chain reaction, protein quantification by means of Western blot, and localization by means of immunofluorescence. We firstly validated HPRT1 as the most reliable housekeeping gene enabling correct gene expression analysis in both granulosa and cumulus cells. Gene expression, Western blot, and immunofluorescence analysis of MMP2, MMP9, and MMP11 and their tissue inhibitors TIMP1 and TIMP2 demonstrated that these enzymes are finely tuned in these cells. MMP9 is specifically expressed only in granulosa, whereas MMP2 is more expressed in cumulus and granulosa cells in cases of reduced ovarian response and decreased fertilization rate. This study sheds light on MMP and TIMP expression in granulosa and cumulus cells, and it may help in understanding the fine regulation of oocyte maturation inside the follicle. Although further studies are needed to fully understand the molecular mechanisms involved in these processes, our findings may be useful in the identification of biomarkers of oocyte maturation, competence acquiring, and fertilization.//////////////////
Matrix metalloproteinases and their inhibitors in human cumulus and granulosa cells as biomarkers for oocyte quality estimation. Luddi A et al. (2018) To study the molecular profile of metalloproteinases and their tissue inhibitors in granulosa and cumulus cells in a subset of fertile and infertile women. Molecular study with granulosa and cumulus cells. University hospital. Forty-four women undergoing assisted reproductive techniques for female infertility factor, with partners having a normal spermiogram and 15 normally fertile women with male partner affected by severe oligoasthenoteratozoospermia or nonobstructive azoospermia. In vitro fertilization. We investigated gene expression level of metalloproteinases (MMP2, MMP9, MMP11) and their tissue inhibitors (TIMP1, TIMP2) by means of quantitative reverse-transcription polymerase chain reaction, protein quantification by means of Western blot, and localization by means of immunofluorescence. We firstly validated HPRT1 as the most reliable housekeeping gene enabling correct gene expression analysis in both granulosa and cumulus cells. Gene expression, Western blot, and immunofluorescence analysis of MMP2, MMP9, and MMP11 and their tissue inhibitors TIMP1 and TIMP2 demonstrated that these enzymes are finely tuned in these cells. MMP9 is specifically expressed only in granulosa, whereas MMP2 is more expressed in cumulus and granulosa cells in cases of reduced ovarian response and decreased fertilization rate. This study sheds light on MMP and TIMP expression in granulosa and cumulus cells, and it may help in understanding the fine regulation of oocyte maturation inside the follicle. Although further studies are needed to fully understand the molecular mechanisms involved in these processes, our findings may be useful in the identification of biomarkers of oocyte maturation, competence acquiring, and fertilization.//////////////////
Immunohistochemical expression of MMP-14 and MMP-2, and MMP-2 activity during human ovarian follicular development. Vos MC 2014 et al.
BACKGROUND
The aim of this study was to investigate the presence of MMP-14 and MMP-2 during human ovarian follicular development using immunohistochemistry, and the activity of MMP-2 in follicular fluid using zymography.
METHODS
Ovarian tissue collected from the archives of the Department of Pathology was examined and medical records and histopathology were reviewed. Follicular fluids were collected at the IVF-department and analyzed using zymography.
RESULTS
MMP-14 and MMP-2 were increasingly found in the growing follicles and MMP-2 was highly expressed in the corpus luteum. Pro-MMP-2 was present in follicular fluid of IVF-patients.
CONCLUSIONS
The presence of MMP-14 and MMP-2 during human ovarian follicular development from the primordial follicle to the tertiary follicle and corpus luteum is confirmed, as was indicated by earlier animal studies following stimulation with gonadotrophins.
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Liu et al examined the physiological regulation pattern and cellular distribution of messenger RNAs coding for gelatinase A (MMP-2) 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.
Characterization of Matrix Metalloproteinase-2 and -9 and their inhibitors in equine granulosa cells in vivo and in vitro. Sessions DR et al. Matrix metalloproteinases (MMP) and tissue inhibitors of MMP (TIMP) regulate tissue remodeling events necessary for ovulation. Thus, changes in MMP and TIMP expression and protein enzyme activity were examined in vivo and in vitro during follicular development and atresia in the horse. Equine granulosa cells and follicular fluid from medium (15 to 29 mm) healthy and atretic follicles and from large (>30 mm) healthy and preovulatory follicles were collected by transvaginal aspiration. The cells were either snap frozen (in vivo study) or cultured for 48 h (in vitro study) to determine gene expression and protein enzyme activity of MMP-2 and -9 and TIMP-1 and -2. Concentrations of progesterone and estradiol were determined by RIA in follicular fluid and conditioned media and were used along with follicle dynamics to classify follicles. In vivo, expression of MMP-2 and TIMP-2 was increased (P < 0.05) in large preovulatory follicles, while TIMP-1 was decreased. The ratio of MMP-2:TIMP-2 expression was decreased (P < 0.05) in medium healthy and large preovulatory follicles, while MMP-9:TIMP-1 ratio was increased only in large preovulatory compared to large healthy follicles. Estradiol was greatest (P < 0.05) in fluid of large healthy and large preovulatory follicles. However, medium atretic follicles were associated with the lowest estradiol concentrations, both in vivo and in vitro. Progesterone concentrations were greatest (P < 0.05) in large preovulatory follicles both in vivo and in vitro. In healthy follicles in vivo, the diameter correlated to estradiol concentration, estradiol:progesterone ratio, MMP-9 and TIMP-1 expression, and MMP-2 and -9 protein activity. In contrast to in vivo studies, the ratio of MMP-9:TIMP-1 expression was increased (P < 0.05) in medium healthy follicles; TIMP-2 expression decreased in large preovulatory follicles in vitro. In addition, MMP-9 protein activity was decreased (P < 0.05) in the media samples of cells from large healthy compared to medium healthy follicles. These results indicate changes in MMP-2 and -9 activities may be essential to the tissue reorganization necessary for ovulation in the equine ovary.
Matrix Metalloproteinases 2 and 9 and their Tissue Inhibitors in the Follicular Fluid of Patients with Polycystic Ovaries Undergoing In Vitro Fertilisation. Baka S et al. The present study was undertaken to investigate the levels of matrix metalloproteinase (MMP)-2, MMP-9 and their tissue inhibitors (TIMP-2 and TIMP-1, respectively) in the follicular fluid of 39 patients with polycystic ovary syndrome (PCOS) and compare them with the levels found in 56 age- and weight-matched normally ovulating women, all undergoing in vitro fertilisation (IVF) treatment. Significantly higher levels of MMP-2 and MMP-9 (p=0.02 and p<0.001, respectively) as well as TIMP-2 and TIMP-1 (p=0.006 and p<0.001, respectively) were found in the PCOS group compared to controls. Women who achieved pregnancy had higher TIMP-1 levels compared to the non-pregnant ones in the control group (p=0.01). In conclusion, women with PCOS exhibited significantly increased gelatinolytic activity compared with controls of similar age and body mass index, thus indicating a more intense extracellular matrix remodelling in this group of patients during IVF treatment due to multiple follicular development and cyst formation.
Follicle stages
Antral, Preovulatory, Corpus luteum
Comment
Shalev E, et al reported the balance between MMP-9 and MMP-2 and their tissue inhibitor
(TIMP)-1 in luteinized granulosa cells and comparison between
women with PCOS and normal ovulatory women.
They assessed the differences in production of MMP-2, MMP-9 and TIMP-1 by
cultured luteinized granulosa cells from women with PCOS and normal ovulatory
women after ovarian stimulation for IVF treatment. In follicular fluid from
women with PCOS, levels of MMP-9 and MMP-2 were higher than the normal group,
as was the basal production of these proteins by cultured cells. Basal
production of TIMP-1 by cultured cells was not different between PCOS and
normal groups. A time-dependent increase in the production of MMP-9 was
observed in cells from both normal and PCOS women, although the increase was
more pronounced in the latter Thus the MMP-TIMP balance is shifted toward
greater MMP activity in luteinized granulosa cells from women with PCOS.
Young KA, Stouffer reported the gonadotropin and Steroid Regulation of Matrix Metalloproteinases and Their Endogenous Tissue Inhibitors in the Developed Corpus Luteum of the Rhesus Monkey During the Menstrual Cycle.
The factors regulating the dynamic expression of matrix metalloproteinases (MMPs) and their inhibitors, TIMPs in the primate corpus luteum (CL) during the menstrual cycle are unknown. The authors hypothesized that luteinizing hormone (LH) or progesterone (P) regulate interstitial-collagenase (MMP-1), the gelatinases (MMP-2, MMP-9), TIMP-1 and TIMP-2 in the CL. Hormone ablation/replacement was performed in rhesus monkeys on luteal d9-11 in five treatment groups (n=4/group): control (no treatment); antide (A; GnRH antagonist); antide +LH; antide +LH +trilostane (TRL; 3betaHSD inhibitor); antide +LH+TRL+R5020 (nonmetabolizable progestin). On d12, the CL was removed with RNA and protein isolated for real-time PCR and immunoassays, respectively. MMP-1 mRNA increased 20-fold with A, whereas LH-replacement maintained MMP-1 mRNA at control levels. Likewise, TRL increased MMP-1 mRNA 54-fold and R5020 prevented this effect. Immunodetectable MMP-1 protein also increased with antide or TRL; these increases were abated with LH or R5020. Gelatinase mRNA and/or protein levels increased with antide (e.g., 3-fold, MMP-2 mRNA) and LH-replacement reduced protein levels (e.g., 11-fold, MMP-2). TRL increased MMP-9 but not MMP-2 expression; however R5020 replacement had no effect on mRNA or protein levels. LH treatment increased TIMP-1 and -2 mRNA and TIMP-1 protein expression compared to controls, and antide groups, whereas R5020 enhanced only immunodetectable TIMP-1. These data strongly suggest that LH suppresses MMP-1 in the primate CL via P. LH also suppresses gelatinases, whether at the mRNA (MMP-2) or protein (MMP-2, -9) levels, perhaps in part via steroids, including P. In contrast, LH promotes TIMP expression, perhaps via steroids including P.
Phenotypes
Mutations
2 mutations
Species: mouse
Mutation name: None
type: null mutation fertility: unknown Comment:Itoh T, et al. reported reduced angiogenesis and tumor progression in gelatinase A-deficient mice.
Species: D. melanogaster
Mutation name: type: null mutation fertility: infertile - ovarian defect Comment: Matrix metalloproteinase 2 is required for ovulation and corpus luteum formation in Drosophila. Deady LD et al. (2015) Ovulation is critical for successful reproduction and correlates with ovarian cancer risk, yet genetic studies of ovulation have been limited. It has long been thought that the mechanism controlling ovulation is highly divergent due to speciation and fast evolution. Using genetic tools available in Drosophila, we now report that ovulation in Drosophila strongly resembles mammalian ovulation at both the cellular and molecular levels. Just one of up to 32 mature follicles per ovary pair loses posterior follicle cells ("trimming") and protrudes into the oviduct, showing that a selection process prefigures ovulation. Follicle cells that remain after egg release form a "corpus luteum (CL)" at the end of the ovariole, develop yellowish pigmentation, and express genes encoding steroid hormone biosynthetic enzymes that are required for full fertility. Finally, matrix metalloproteinase 2 (Mmp2), a type of protease thought to facilitate mammalian ovulation, is expressed in mature follicle and CL cells. Mmp2 activity is genetically required for trimming, ovulation and CL formation. Our studies provide new insights into the regulation of Drosophila ovulation and establish Drosophila as a model for genetically investigating ovulation in diverse organisms, including mammals.//////////////////