Remodeling and relaxation of the mouse pubic symphysis (PS) are central events in parturition. The involvement of endogenous proteins such as matrix metalloproteinases (MMPs), tissue inhibitors of matrix metalloproteinases (TIMPs), and cathepsins in these phenomena remains unclear. In this work, we used a combination of immunolocalization, protein expression/activity, and relative messenger RNA (mRNA) expression to examine the changes in selected MMPs (-2, -9, and -8), TIMPs (-1 and -2), and cathepsins (B and K) during pregnancy and postpartum in mice. Immunohistochemistry revealed the presence of all of these proteins in the cytoplasm of chondrocytes, fibrochondrocytes, and fibroblast-like cells in the interpubic tissues. Zymography showed increases in the active forms of MMP-2 and -9 primarily on days 15 to 19 of pregnancy. Western blotting showed enhanced expression of MMP-8 on days 12 to 15 of pregnancy, with no changes in cathepsins B and K. Matrix metalloproteinases 2, TIMP-1 and -2, and cathepsin B had significant relative gene expression throughout pregnancy. These findings indicate that during pregnancy and postpartum there are variations in the expression and activity of proteins that may have an important role in remodeling the pubic symphysis during these events.
HallK. The effects of pregnancy and relaxin on the histology of the pubic symphysis in the mouse. J Endocrinol. 1947;5(3):174–182.
2.
CrelinES. The development of the bony pelvis and its changes during pregnancy and parturition. Trans N Y Acad Sci. 1969;31(8):1049–1058.
3.
CrelinESNewtonEV. The pelvis of the free-tailed bat: sexual dimorphism and pregnancy changes. Anat Rec. 1969;164(3):349–357.
4.
WahlLMBlandauRJPageRC. Effect of hormones on collagen metabolism and collagenase activity in the pubic symphysis ligament of the guinea pig. Endocrinology. 1977;100(2):571–579.
5.
GambleJGSimmonsSCFreedmanM. The symphysis pubis. Anatomic and pathologic considerations. Clin Orthop Relat Res. 1986;203(2):261–272.
6.
SherwoodOD. RelaxinIn. KnobilENeillJD, eds. The Physiology of Reproduction. vol 2. New York, NY: Raven Press; 1994:861–1009.
7.
HashemGZhangQHayamiTChenJWangWKapilaS. Relaxin and β-estradiol modulate targeted matrix degradation in specific synovial joint fibrocartilages: progesterone prevents matrix loss. Arthritis Res Ther. 2006;8(4):R98.
SnowRENeubertAG. Peripartum pubic symphysis separation: a case series and review of the literature. Obstet Gynecol Surv. 1997;52(7):438–443.
10.
BjorklundKBergströmSNordströmMLUlmstenU. Symphyseal distention in relation to serum relaxin levels and pelvic pain in pregnancy. Acta Obstet Gynecol Scand. 2000;79(4):269–275.
11.
OwensKPearsonAMasonG. Symphysis pubis dysfunction––a cause of significant obstetric morbidity. Eur J Obstet Gynecol Reprod Biol. 2002;105(2):143–146.
12.
ZhaoLSamuelCSTregearGWBeckFWintourEM. Collagen studies in late pregnant relaxin null mice. Biol Reprod. 2000;63(3):697–703.
13.
TalmageRV. Changes produced in the symphyseal pubis of guinea pig by the sex steroids and relaxin. Anat Rec. 1947;99(1):91–113.
14.
TalmageRV. A histological study of the effects of relaxin on the symphysis pubis of the guinea pig. J Exp Zool. 1947;106(3):281–297.
15.
McDonaldJKSchwabeC. Relaxin induced elevations of cathepsin B and dipeptidyl peptidase I in the mouse pubic symphysis, with localization by fluorescence enzyme histochemistry. Ann N Y Acad Sci.1982;380:178–186.
16.
OrtegaHHMuñoz-de-ToroMMLuqueEHMontesGS. Morphological characteristic of the interpubic joint (symphysis pubica) of rats, guinea pigs and mice in different physiological situations. A comparative study. Cells Tissues Organs. 2003;173(2):105–114.
17.
MoraesSGPinheiroMCToledoOMSJoazeiroP. Phenotypic modulation of fibroblastic cells in mice pubic symphysis during pregnancy, partum and postpartum. Cell Tissue Res. 2004;315(2):223–231.
18.
VeridianoAMGarciaEAPinheiroMCNishimoriFYToledoOMSJoazeiroPP. The mouse pubic symphysis as a remodeling system: morphometrical analysis of proliferation and cell death during pregnancy, partus and postpartum. Cell Tissue Res. 2007;330(1):161–167.
19.
WeissMNagelschmidtMStruckH. Relaxin and collagen metabolism. Hormone Metab Res. 1979;11(6):408–410.
20.
RosaRGTarsitanoCAHyslopSYamadaATToledoOMJoazeiroPP. Relaxation of the mouse pubic symphysis during late pregnancy is not accompanied by the influx of granulocytes. Microsc Res Tech. 2008;71(3):169–178.
21.
PinheiroMCJoazeiroPPMoraOAToledoOM. Ultrastructural and immunohistochemistry analysis of proteoglycans in mouse pubic symphysis. Cell Biol Int. 2003;27(8):647–655.
22.
PinheiroMCMoraOACaldiniEGBattlehnerCNJoazeiroPPToledoOMS. Ultrastructural, immunohistochemical and biochemical analysis of glycosaminoglycans and proteoglycans in the mouse pubic symphysis during pregnancy. Cell Biol Int. 2005;29(6):458–471.
23.
SomervilleRPTOblanderSAApteSS. Matrix metalloproteinase: old dogs with new tricks. Genome Biol. 2003;4(6):216.01–216.11.
24.
McCawleyLJMatrisianLM. Matrix metalloproteinases: they’re not just for matrix anymore!Curr Opin Cell Biol. 2001;13(5):534–540.
25.
LenhartJARyanPTOhlethKMPalmerSSBagnellCA. Relaxin increases secretion of tissue inhibitor of matrix metalloproteinase-1 and -2 during uterine and cervical growth and remodelling in the pig. Endocrinology. 2002;143(1):91–98.
26.
StygarDWangHVladicYSEkmanGErikssonHSahlinL. Increased level of matrix metalloproteinases 2 and 9 in the ripening process of the human cervix. Biol Reprod.2002;67(3):889–894.
27.
NuttallRKSampieriCLPenningtonCJGillSESchultzGAEdwardsDR. Expression analysis of the MMP and TIMP gene families during mouse tissue development. FEBS Lett. 2004;563(1-3):129–134.
28.
CurryTEOsteenKG. The matrix metalloproteinase system: changes, regulation, and impact throughout the ovarian and uterine reproductive cycle. Endocrine Rev. 2003;24(4):428–465.
29.
ImadaKItoASatoTNamikiMNagaseHMoriY. Hormonal regulation of matrix metalloproteinase 9/gelatinase B gene expression in rabbit uterine cervical fibroblasts. Biol Reprod. 1997;56(3):575–580.
30.
UusitaloHHiltunenASoderstromMAroHTVuoroE.Expression of cathepsins B, H, K, L, and S and matrix metalloproteinases 9 and 13 during chondrocyte hypertrophy and endochondral ossification in mouse fracture callus. Calcif Tissue Int. 2000;67(5):382–390.
31.
SennströmMBBraunerABystromBMalmstromAEkmanG. Matrix metalloproteinase-8 correlates with the cervical ripening process in humans. Acta Obstet Gynecol Scand. 2003;82(10):904–911.
32.
NodaMOhJTakahashiRKondoSKitayamaHTakahashiC. RECK: a novel suppressor of malignancy linking oncogenic signaling to extracellular matrix remodeling. Cancer Metastasis Rev. 2003;22(2-3):167–175.
33.
ChircoRLiuXWJungKKKimRRC. Novel functions of TIMPs in cell signalling. Cancer Metastasis Rev. 2006;25(1):99–113.
34.
AmălineiCCăruntuIDBălanRA. Biology of metalloproteinases. Roman J Morphol Embryol. 2007;48(4):323–334.
35.
HulboyDlRudolphLAMatrisianLM. Matrix metalloproteinases as mediators of reproductive function. Mol Human Rep. 1997;3(1):27–45.
36.
LecailleFBrommeDLalmanachG. Biochemical properties and regulation of cathepsin K activity. Biochimie. 2008;90(2):208–226.
37.
LaemmliUK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680–685.
38.
CalvoIROcónBMoyaPMReversible Ponceau staining as a loading control alternative to actin in western blots. Anal Biochem. 2010;401(2):318–320.
39.
DiScipioRGSchraufstatterIUSikoraLZurawBLSriramaraoP. C5a mediates secretion and activation of matrix metalloproteinase 9 from human eosinophils and neutrophils. Int Immunopharmacol. 2006;6(7):1109–1118.
40.
LabordaJ. 36B4 cDNA used as an estradiol-independent mRNA control is the cDNA for human acidic ribosomal phosphoprotein PO. Nucleic Acids Res. 1991;19(14):3998.
41.
StamenkovicI. Extracellular matrix remodelling: the role of matrix metalloproteinases. J Pathol. 2003;200(4):448–464.
NaesseEPSchreursOHelgelandKSchenckKSteinsvollSM. Matrix metalloproteinases and their inhibitors in gingival mast cells in persons with and without human immunodeficiency virus infection. J Periodontol. 2003;38(6):575–582.
44.
ParryLJMcGuaneJTGehringHMKosticIGTSiebelAL. Mechanisms of relaxin action in the reproductive tract: studies in the relaxin-deficient (Rlx-/- mouse. Ann N Y Acad Sci.20051041:91–103.
45.
KerkhofMHHendriksHBrölmannHA. Changes in connective tissue in patients with pelvic organ prolapse—a review of the current literature. Int Urogynecol J. 2009;20(4):461–474.
46.
DuboisBArnoldBOpdenakkerG. Gelatinase B deficiency impairs reproduction. J Clin Invest. 2000;106(5):627–628.
47.
Page-McCawAEwaldAJWerbZ. Matrix metalloproteinases and the regulation of tissue remodeling. Nat Rev Mol Cell. 2007;8(3):221–233.
48.
LambertVWielockxBMunautCMMP-2 and MMP-9 synergize in promoting choroidal neovascularization. FASEB J. 2003;17(15):2290–2292.
49.
YangSRembiesaBBüllesbachEESchwabeC. Relaxin receptors in mice: demonstration of ligand binding in symphyseal tissues and uterine membrane fragments. Endocrinology. 1992;130(1):179–185.
50.
LenhartJARyanPTOhlethKMPalmerSSBagnellCA. Relaxin increases secretion of matrix metalloproteinase-2 and matrix metalloproteinase-9 during uterine and cervical growth and remodeling in the pig. Endocrinology. 2001;142(9):3941–3949.
51.
ArguelloRJPérezCEDelgadoCRSolorzaLGVillaTSArenasHF. Matrix metalloproteinases-2, -3, and -9 secreted by explants of benign and malignant lesions of the uterine cervix. Int J Gynecol. 2004;14(2):333–340.
52.
HennemanSBildtMMDeGrootJKuijpers-JagtmanAMVon den HoffJW. Relaxin stimulates MMP-2 and α-smooth muscle actin expression by human periodontal ligament cells. Arch Oral Biol. 2008;53(2):161–167.
53.
WangWHayamiTKapilaS. Female hormone receptors are differentially expressed in mouse fibrocartilages. Osteoarthr Cartil. 2009;17(5):646–654.
54.
AnBSChoiKCKangSKMouse calbindin-D9k gene expression in the uterus during late pregnancy and lactation. Mol Cell Endocrinol. 2003;205(1-2):79–88.
55.
BrewKDinakarpandianDNagaseH. Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochim Biophys Acta. 2000;1477(1-2):267–283.
56.
FassinaGFerrariNBrigatiCTissue inhibitors of metalloproteinases: regulation and biological activities. Clin Exp Metastasis. 2000;18(2):111–120.
57.
MasonRWJonsonDABarretAJChapmanHA. Elastinolytic activity of human cathepsin L. Biochem J. 1986;233(3):925–927.
58.
KostoulasGLangANagaseHBaiciA. Stimulation of angiogenesis through cathepsin B inactivation of the tissue inhibitors of matrix metalloproteinases. FEBS J. 1999;455(3):286–290.
59.
TimmonsBAkinsMMahendrooM. Cervical remodeling during pregnancy and parturition. Trends Endocrinol Metab. 2010;21(6):353–361.
60.
WieslanderCKMarinisSIDrewesPGKellerPWAcevedoJFWordAR. Regulation of elastolytic proteases in the mouse vagina during pregnancy, parturition, and puerperium. Biol Reprod. 2008;78(3):521–528.
