Research articleExpression and localization of matrix metalloproteinases (MMP-2, -7, -9) and their tissue inhibitors (TIMP-2, -3) in the chicken oviduct during pause in laying induced by tamoxifen
Introduction
The avian oviduct has been recognized as an outstanding model for investigating the biological actions of estrogen [1]. In the chicken oviduct, estrogen induces cell proliferation and differentiation and exhibits antiapoptotic effects [2], [3]. Besides stimulation of the oviduct growth, estrogen induces synthesis of several egg proteins in the oviduct and liver due to both transcriptional increase and mRNA stabilization [1], [4].
The physiological actions of estrogen are mediated by its nuclear receptors, estrogen receptor alfa, and estrogen receptor beta, although evidence of rapid cytoplasmic responses to estrogen is also increasing. These receptors function primarily as ligand-activated transcription factors and bind directly with target genes at estrogen response elements or indirectly via tethering to the AP-1 or Sp1 transcription factors. Cytoplasmic signaling pathways can also activate estrogen receptor (ER) in a ligand-dependent or ligand-independent manner by phosphorylation of ER directly or by phosphorylation of its coactivators. The changes in conformation of ER caused by ligands likely determine its interaction with specific coactivators or corepressors resulting in the stimulation or inhibition of transcription at a given promoter site of an estrogen-responsive gene [1].
The triphenylethylene antiestrogens, including tamoxifen (TMX), are very potent antagonists of estrogen action in the chicken and manifest little agonist activity compared to their action in other species. TMX is thought to bind to ER receptors and work against the effect of estrogen in the target tissue [5], [6]. Therefore, it has been frequently used to investigate estrogen action in birds. This study was designed to evaluate the influence of TMX treatment on the expression, localization and activity of selected matrix metalloproteinase (MMP) system members in the chicken oviduct.
The MMP comprise a family of proteolytic enzymes that include four broad classes: the collagenases, gelatinases, stromelysins, and membrane-type enzymes. The gelatinases (MMP-2 and MMP-9) contain a fibronectin-like sequence within their catalytic domain, which results in a potent ability for these MMPs to cleave gelatin. The stromelysin enzymes (e.g., MMP-7) act on a broad and diverse array of extracellular matrix (ECM) substrates. Both the gelatinases and stromelysins are capable of degrading major constituents of basement membranes, including type IV collagen, laminin, and fibronectin. In addition to degrading the ECM, the MMPs and especially the stromelysins exhibit activity toward other MMPs, growth factors, cytokines, chemokines, and other growth regulators that are matrix bound or present on the cell surface [7], [8].
Expression of MMPs is controlled at the level of transcription in response to proto-oncogenes and exogenous signals, including hormones. At the posttranslational level, MMPs are regulated by both serum-borne and tissue-derived endogenous inhibitors termed tissue inhibitors of metalloproteinases (TIMPs). TIMPs are widely expressed in all tissues, and they are regulated jointly with the MMPs in most cases. In addition to inhibition of MMPs, the TIMPs participate in several essential biological functions. For example, TIMP-2 stimulates fibroblast growth [9], and TIMP-3 induces apoptosis and regulates steroidogenesis and growth of ovarian follicles [10]. MMP-2 is primarily regulated by TIMP-2, whereas MMP-9 is regulated by TIMP-3 [11]. Furthermore, most MMPs are secreted as zymogens, requiring the cleavage of a specific propeptide domain for activation. However, the mechanisms orchestrating pro-MMP activation have not been fully elucidated.
The activity of MMPs in the reproductive system of mammals is regulated by the sex steroids, growth factors, and cytokines that control reproductive function [12], [13], [14]. However, despite the fact that the MMP system has been described in the mammalian oviduct and uterus [14], [15], [16], [17], its role and regulation in the avian oviduct is poorly understood. First, the mRNAs of MMP-2, TIMP-2, and TIMP-3 were found in the chicken oviduct by Réhault-Godbert et al. [18], and our previous studies revealed changes in the expression and activity of some MMP system members in the chicken oviduct during sexual maturation [19] as well as during pause in egg laying induced by food deprivation [20]. In the current study, our aim was to investigate the expression and localization of selected MMPs (MMP-2, MMP-7, MMP-9) and TIMPs (TIMP-2, TIMP-3) as well as MMPs activity in the particular segments of the chicken oviduct during pause in laying induced by TMX (ER modulator) treatment. Additionally, concentration of ovarian steroids was measured in blood plasma.
Section snippets
Birds and experimental design
The animal experiment was conducted according to research protocol approved by the Local Animal Ethics Committee in Kraków, Poland. Laying Hy-Line Brown hens were purchased from a commercial farm, H&P2 s.c. (Czarków, Poland). Chickens were caged individually under a photoperiod of 14L:10D with commercial food and water ad libitum.
The 12 laying hens at 25 weeks of age (average body weight 1.86 ± 0.036) were divided into two groups. The experimental chickens (n = 6) were treated daily (for
Changes in the oviduct weight after TMX treatment
On day six of the experiment the hens treated with TMX entirely stopped egg laying.
The weight of the oviduct in the TMX-treated chickens was 17% lower than in the control hens (P < 0.001).
Sex hormones concentrations after TMX treatment
Concentrations of steroids in the blood plasma of control and experimental chickens are shown in Figure 1A–C. Treatment with TMX significantly decreased the progesterone level (by 66% on Day 7 of experiment in comparison with control), while it did not influence testosterone and estradiol concentrations.
Messenger RNA expression of MMPs and TIMPs in the chicken oviduct after TMX treatment
Discussion
Alterations in the expression of MMPs and TIMPs and activity of MMPs in the chicken oviduct during sexual maturation and pause in egg laying observed in previous studies [19], [20] are accompanied by changes in the concentrations of steroid hormones in the blood plasma of birds [22], [23], [24], [25], which may indicate the involvement of steroids in the regulation of MMP system components. The main hormones that control the development and functioning of the oviduct are estrogens, which
Acknowledgments
This work was financially supported by the National Science Center, Poland (grant PRELUDIUM No. UMO-2012/07/N/NZ4/00165 dedicated to A.L.-W). Results presented in part at the 18th Annual Conference of the European Society for Domestic Animal Reproduction (ESDAR), September 11–13 Helsinki, Finland.
References (57)
- et al.
Estrogen action: revitalization of the chick oviduct model
Trends Endocrinol Metab
(2005) - et al.
Estrogen-responsive genes encoding egg yolk proteins vitellogenin and apolipoprotein II in chicken are differentially regulated by selective estrogen receptor modulators
Theriogenology
(2016) Interactions of tamoxifen in the chicken
J Steroid Biochem
(1987)- et al.
Extracellular regulation of metalloproteinases
Matrix Biol
(2015) - et al.
Activation of a recombinant membrane type 1-matrix metalloproteinase (MT1-MMP) by furin and its interaction with tissue inhibitor of metalloproteinases (TIMP)-2
FEBS Lett
(1996) - et al.
Expression and regulative function of tissue inhibitor of metalloproteinase 3 in the goat ovary and its role in cultured granulosa cells
Mol Cell Endocrinol
(2015) - et al.
Electrophoretic transfer protein zymography
Anal Biochem
(2011) - et al.
Exogenous leptin advances puberty in domestic hen
Domest Anim Endocrinol
(2006) - et al.
Effect of progesterone in vitro on luteinizing hormone production in hen pituitary cells pretreated with estrogen
Poult Sci
(1992) - et al.
Localization of progesterone receptors in the shell gland of laying and nonlaying chickens
Poult Sci
(1991)
Immunohistochemical localization of progesterone receptor isoforms and estrogen receptor alpha in the chicken oviduct magnum during development
Acta Histochem
Changes in progesterone receptor isoforms expression and in the morphology of the oviduct magnum of mature laying and aged nonlaying hens
Biochem Biophys Res Commun
Changes in mRNA expression of MMP-2 in the Müllerian duct of chicken embryo
Gen Comp Endocrinol
The regulation of matrix metalloproteinases and their inhibitors
Int J Biochem Cell Biol
Prostaglandin E2 stimulates expression of matrix metalloproteinase 2 in cultured rat mesangial cells
Kidney Int
EMMPRIN, an upstream regulator of MMPs, in CNS biology
Matrix Biol
Cytokines in reproductive remodeling of molting White Leghorn hens
J Reprod Immunol
Estrogen-induced cytodifferentiation of the ovalbumin-secreting glands of the chick oviduct
J Cell Biol
Tissue-protective effects of estrogen involve regulation of caspase gene expression
Mol Endocrinol
Estrogen antagonists in chick oviduct: antagonist activity of eight synthetic triphenylethylene derivatives and their interactions with cytoplasmic and nuclear estrogen receptors
Endocrinology
Matrix metalloproteinases and the regulation of tissue remodeling
Nat Rev Mol Cell Biol
The matrix metalloproteinase system: changes, regulation, and impact throughout the ovarian and uterine reproductive cycle
Endocr Rev
Cyclic changes in the matrix metalloproteinase system in the ovary and uterus
Biol Reprod
Differential expression of extracellular matrix components in the bovine oviduct during the oestrous cycle
Reproduction
Regulated expression of matrix metalloproteinases, inflammatory mediators, and endometrial matrix remodeling by 17beta-estradiol in the immature rat uterus
Reprod Biol Endocrinol
Regulation of MMP-9 expression and activity in the mouse uterus by estrogen
Mol Reprod Dev
Estrogen suppresses expression of the matrix metalloproteinase inhibitor reversion-inducing cysteine-rich protein with Kazal motifs (RECK) within the mouse uterus
Endocrine
Detection of the matrix metalloproteinases MMP-2 and MMP-9 and tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2 in llama (Lama glama) oviduct
Reprod Domest Anim
Cited by (14)
Alterations in connexin 43 gene and protein expression in the chicken oviduct following tamoxifen treatment
2022, TheriogenologyCitation Excerpt :Hence, we verify whether the estrogen action blockage will modify the Cx43 expression and localization in the hen oviduct. The TMX treatment of birds was applied since it is established that in the chicken oviduct, TMX acts as an antagonist of the estrogen receptors and causes oviduct regression and a pause in egg laying [9,23,43]. The results of the present study confirm the earlier records and the discovered lessening in Cx43 expression at the mRNA level in the magnum and isthmus and at the protein level in the isthmus and vagina of TMX-treated hens compared with the control chickens.
Reproduction in the female
2022, Sturkie's Avian PhysiologyTamoxifen-induced alterations in the expression of selected matrix metalloproteinases (MMP-2, -9, -10, and −13) and their tissue inhibitors (TIMP-2 and -3) in the chicken ovary: MMPs in chicken ovary following tamoxifen treatment
2020, TheriogenologyCitation Excerpt :For this purpose, the laying hens were treated with TMX, to block the estrogen receptors, and in consequence estrogen action. Treatment of chickens with TMX over several days leads to complete cessation of egg laying and regression of the ovary and oviduct [25–27,35]. These effects of estrogen action deficiency following TMX treatment were also confirmed in this study.