Abstract
The epidermal growth factor (EGF) plays a crucial role in the control of uterine cell proliferation, growth and differentiation. This study was designed to investigate the spatiotemporal expression pattern and localization of the EGF receptor/ligand system during the process of uterine involution using immunohistochemistry. Our results indicated that the expression of the ErbB/HER receptors and their ligands varied with structural changes in the uterus at different days of involution. Supranuclear punctate ErbB1 immunostaining was observed in the luminal and glandular epithelial cells and endometrial fibroblasts. Moderate ErbB2/HER2 immunoreactivity was observed in the lateral membrane and cytoplasm of the epithelial cells on the 1st, 3rd and 5th days and was decreased on the other days of involution. The amount of nuclear and cytoplasmic ErbB3/HER3 and ErbB4/HER4 immunostaining remained constant throughout the postpartum period. The EGF immunoreaction was weak in the luminal and glandular epithelium throughout the involution period. Although the cytoplasmic AREG immunoreactivity in the glandular epithelium was stronger on the 1st and 3rd days compared with the other days of involution, NRG1 immunostaining was weak on the 1st and 3rd days and was moderate in the apical cytoplasm on the 10th and 15th days of involution. The macrophages displayed strong cytoplasmic immunoreactivity for ErbB3/HER3, ErbB4/HER4, EGF, AREG and NRG. Strong, moderate and weak immunostaining for ErbB2/HER2, ErbB4/HER4 and other proteins (ErbB1, ErbB3, AREG and NRG), respectively, was present in the myometrial smooth muscle cells. These findings support the hypothesis that the EGFsystem plays a role in the development of various physiological changes associated with uterine involution.
Similar content being viewed by others
References
Afting E-G, Elce JS (1978) DNA in the rat uterus myometrium during pregnancy and postpartum involution: Measurement of DNA in small pieces of mammalian tissue. Anal Biochem 86:90–99
Aghajanova L, Bjuresten K, Altmae S, Landgren BM, Stavreus-Evers A (2008) HB-EGF but not amphiregulin or their receptors HER1 and HER4 is altered in endometrium of women with unexplained infertility. Reprod Sci 15:484–492
Aker ON (1954) Laboratuvar El Kitabı, Hususi Boyama Teknikleri. Örnek Matbaası, Ankara. S:138–139
Alan E, Liman N (2012) Immunohistochemical localization of beta defensins in the endometrium of rat uterus during the postpartum involution period. Vet Res Commun 36:173–185
Andrechek ER, Hardy WR, Girgis-Gabardo AA et al (2002) ErbB2 is required for muscle spindle and myoblast cell survival. Mol Cell Biol 22:4714–4722
Bazley LA, Gullick WJ (2005) The epidermal growth factor receptor family. Endocr Relat Cancer 12:17–27
Blackburn ST (2007) Maternal, fetal, and neonatal physiology: a clinical perspective. Saunders Elseiver, St. Louis Missouri, p 153
Boomsma RA, Mavrogianis PA, Verhage HG (1997) Immunohistochemical localization of TGF-α, EGF and EGFR in the cat endometrium and placenta. Histochem J 29:495–504
Brown NM, Lamartiniere CA (2000) Genistein regulation of transforming growth factor-a, epidermal growth factor (EGF), and EGF Receptor expression in the rat uterus and vagina. Cell Growth Differ 11:255–260
Brown N, Deb K, Paria BC, Das SK, Reese J (2004) Embryo-uterine interactions via the neuregulin family of growth factors during implantation in the mouse. Biol Reprod 71:2003–2011
Burden S, Yarden Y (1997) Neuregulins and their receptors: a versatile signaling module in organogenesis and oncogenesis. Neuron 18:847–855
Calvo M, Zhu N, Tsantoulas C, Ma Z, Grist J, Loeb JA, Bennett DL (2010) Neuregulin–erbB signaling promotes microglial proliferation and chemotaxis contributing to microgliosis and pain after peripheral nerve injury. J Neurosci 30:5437–5450
Carpenter RL, Lo H-W (2013) Regulaton of apoptosis by HER2 in breast cancer. J Carcinog Mutagen S7:003
Chegini N, Rossi M, Masterson B (1992) Platelet-derived growth factor (PDGF), epidermal growth factor (EGF), and EGF and PDGF beta-receptors in human endometrial tissue: localization and in vitroaction. Endocrinology 130:2373–2385
Chobotova K, Karpovich N, Carver J et al (2005) Heparin-binding epidermal growth factor and its receptors mediate decidualization and potentiate survival of human endometrial stromal cells. J Clin Endocrinol Metab 90:913–919
Christensen ST, Pedersen SF, Satir P, Veland IR, Scneider L (2008) The primary cilium coordinates signaling pathways in cell cycle control and migration during development and tissue repair. Curr Top Dev Biol 85:261–301
Clark MV, Brinsfield TH (1973) Ciliated stromal cells in the hamster uterus. Biol Reprod 8:100–102
Crossman G (1937) A modification of Mallory’s connective tissue stain with a discussion of the principles involved. Anat Rec 69:33–38
Das SK, Tsukamura H, Paria BC, Andrews GK, Dey SK (1994) Differantial expression of epidermal growth factor receptor (EGF-R) gene and regulation of EGF-R bioactivity by progestrone and estrogen in the adult mouse uterus. Endocrinology 134:971–981
Das SK, Chakraborty I, Paria BC, Wang XN, Plowman G, Dey SK (1995) Amphiregulin is an implantation-specific and progesterone-regulated gene in the mouse uterus. Mol Endocrinol 9:691–705
Das SK, Das N, Wang J et al (1997) Expression of betacellulin and epiregulin genes in the mouse uterus temporally by the blastocyst solely at the site of its apposition is coincident with the “window” of implantation. Dev Biol 190:178–190
DiAugustine RP, Petrusz P, Bell GL et al (1988) Influence of estrogens on mouse uterine epidermal growth factor precursor protein and messenger ribonucleic acid. Endocrinology 122:2355–2363
Ejskjaer K, Sorensen BS, Poulsen SS, Mogensen O, Forman A, Nexo E (2005) Expression of the epidermal growth factor system in human endometrium during the menstrual cycle. Mol Hum Reprod 11:543–551
Esslemont RJ, Kossaibati MA (2000) The use of databases to manage fertility. Anim Reprod Sci 60–61:725–741
Falls DL (2003) Neuregulins: functions, forms, and signaling strategies. Exp Cell Res 284:14–30
Gardner RM, Lingham RB, Stancel GM (1987) Contractions of the isolated uterus stimulated by epidermal growth factor. FASEB J 1:224–228
Gardner RM, Verner G, Kirkland JL, Stancel GM (1989) Regulation of uterine epidermal growth factor (EGF) receptors by estrogen inthe mature rat and during the estrous cycle. J Steroid Biochem 32:339–343
Gerstenberg C, Allen W, Stewart F (1999) Factors controlling epidermal growth factor (EGF) gene expression in the endometrium of the mare. Mol Reprod Dev 53:255–265
Gharib-Hamrouche N, Chene N, Martal J (1995) Comparative expression of TGF-α and EGF genes in the ovine conceptus and uterine endometrium in the peri-implantation period. Reprod Nutr Dev 35:291–303
Glasser SR, Julian J, Decker GL, Tang J-P, Carson DD (1988) Development of morphological and functional polarity in primary cultures of immature rat uterine epithelial cells. J Cell Biol 107:2409–2423
Gui Y, Zhang J, Liang W, Cai Z (2008) Expression of amphiregulin in human endometrium during the menstrual cycle. Beijing Da Xue Xue Bao 40:241–244
Haley JD, Gullick WJ (2008) EGFR Signaling Networks in Cancer Therapy. Humana Press, Springer Science + Business Media, LLC, New York, p 193
Harrison-Woolrych ML, Charnock-Jones DS, Smith SK (1994) Quantification of messenger ribonucleic acid for epidermal growth factor in human myometrium and leiomyomata using reverse transcriptase polymerase chain reaction. J Clin Endocrinol Metab 78:1179–1184
Huet-Hudson YM, Chakraborty C, De SK, Suzuki Y, Anderws GK, Dey SK (1990) Estrogen regulates the synthesis of epidermal growth factor in Mouse uterine epithelial cells. Mol Endocrinol 4:510–523
Idris N, Carothers Carraway CA, Carraway KL (2001) Differential localization of erbB2 in different tissues of the rat female reproductive tract: implications for the use of specific antibodies for erbB2 analysis. J Cell Physiol 189:162–170
Imai T, Kurachi H, Adachi K et al (1995) Changes in epidermal growth factor receptor and the levels of its ligands during menstrual cycle in human endometrium. Biol Reprod 52:928–938
Jiang N, Saba NF, Chen ZG (2012) Advances in targeting HER3 as an anticancer therapy. Chemother Res Pract 2012:817304
Kida K, Maezono Y, Kawate N, Inaba T, Hatoya S, Tamada H (2010) Epidermal growth factor, transforming growth factor-α, and epidermal growth factor receptor expression and localization in the canine endometrium during the estrous cycle and in bitches with pyometra. Theriogenology 73:36–47
Kim GY, Besner GE, Steffen CL et al (1995) Purification of heparin-binding EGF-like growth factor from pig uterine luminal flushings, and its production by endometrial tissues. Biol Reprod 52:561–571
Kim JG, Vallet JL, Christenson RK (2003) Molecular cloning and endometrial expression of porcine amphiregulin. Mol Reprod Dev 65:366–372
Klonisch T, Wolf P, Hombach-Klonisch S et al (2001) Epidermal growth factor-like ligands and erbB genes in the peri-implantation rabbit uterus and blastocyst. Biol Reprod 64:1835–1844
Komárek V (2000) Snopsis of the organ anatomy. In: Krinke GJ (ed) The laboratory rat. Academic, San Diego, p 315
Konopka B, Skasko E, Kluska A et al (1998) Changes in the concentrations of receptors of insulin-like growth factor-I, epithelial growth factor, oestrogens and progestagens in adenomyosis foci, endometrium and myometrium of women during menstrual cycle. Eur J Gynaecol Oncol 19:93–97
Koumakpayi IH, Diallo JS, Le Page C et al (2006) Expression and nuclear localization of ErbB3 in prostate cancer. Clin Cancer Res 12:2730–2737
Koumas L, King AE, Critchley HOD, Kelly RW, Phipps RP (2001) Fibroblast heterogeneity: existence of functionally distinct Thy 1+ and Thy 1− human female reproductive tract fibroblasts. Am J Pathol 159:925–935
Lamb DJ, Modjtahedi H, Plant NJ, Ferns G (2004) EGF mediates monocyte chemotaxis and macrophage proliferation and EGF receptor is expressed in atherosclerotic plaques. Atherosclerosis 176:21–26
Lee DS, Ueta YY, Xuan X et al (2005) Expression patterns of the implantation-associated genes in the uterus during the estrous cycle in mice. J Reprod Dev 51:787–798
Lee D-S, Yanagimoto Ueta Y, Suzuki H (2006) Expression of amphiregulin during the pre- and post-implantation period in the mouse reproduction tract. J Reprod Dev 52:781–787
Lennard SN, Gerstenberg C, Allen WR, Stewart E (1998) Expression of epidermal growth factor and its receptor in equine placental tissues. J Reprod Fertil 112:49–57
Lim H, Dey SK, Das SK (1997) Differential expression of the erbB2 gene in the periimplantation mouse uterus: potential mediator of signaling by epidermal growth factor-like growth factors. Endocrinology 138:1328–1337
Lim H, Das N, Dey SK (1998) ErbB genes in the mouse uterus: cell-specific signaling by epidermal growth factor (EGF) family of growth factors during implantation. Dev Biol 204:97–110
Liman N, Alan E, Küçük Bayram G, Gürbulak K (2013) Expression of survivin, Bcl-2 and bax proteins in the domestic Cat (Felis catus) endometrium during the oestrus cycle. Reprod Domest Anim 48:33–45
Lin SY, Makino K, Xia W et al (2001) Nuclear localization of EGF receptor and its potential new role as a transcription factor. Nat Cell Biol 3:802–808
Mısırlıoglu D, Nak D, Sevimli A et al (2006) Steroid receptor expression and HER-2/neu (c-erbB-2) oncoprotein in the uterus of cats with cystic endometrial hyperplasia–pyometra complex. J Vet Med A 53:225–229
Miturski R, Semczuk A, Jacowicki JA (1998) C-erbB-2 expression in human proliferative and hyperplastic endometrium. Int J Gynecol Obstet 61:73–74
Möller B, Rasmussen C, Lindblom B, Olovsson M (2001) Expression ofthe angiogenic growth factors VEGF, FGF-2, EGF and their receptors in normal human endometrium during the menstrual cycle. Mol Hum Reprod 7:65–72
Monney L, Otter I, Olivier R et al (1996) Bcl-2 overexpression blocks activation of the death protease CPP32/Yama/apopain. Biochem Biophys Res Commun 221:340–345
Mukku VR, Stancel GM (1985) Receptors for epidermal growth factor in the rat uterus. Endocrinology 117:149–154
Nelson KG, Takahashi T, Lee DC et al (1992) Transforming growth factor-α is a potential mediator of estrogen actionin the mouse uterus. Endocrinology 131:1657–1664
Niikura H, Sasano H, Kaga K, Sato S, Yajima A (1996) Expression of epidermal growth factor family proteins and epidermal growth factor receptor in human endometrium. Hum Pathol 27:282–289
Offterdinger M, Schofer C, Weipoltshammer K, Grunt TW (2002) c-erbB-3: a nuclear protein in mammary epithelial cells. J Cell Biol 157:929–939
Onagbesan OM, Gullick W, Woolveridge I, Peddie MJ (1994) Immunohistochemical localization of epidermal growth factor receptors, epidermal-growth-factor-like and transforming-growth-factor-alpha-like peptides in chicken ovarian follicles. J Reprod Fertil 102:147–153
Padycula HA (1976) Cellular mechanisms involved in cyclic stromal renewal of the uterus. III. Cells of the immune response. Anat Rec 184:49–72
Parakkal PF (1972) Macrophages: the time course and sequence of their distribution in the postpartum uterus. J Ultrastruct Res 40:284–291
Petraglia F, Strauss JF, Weiss G, Gabbe SG (2007) Preterm birth: mechanisms, mediators, prediction, prevention & interventions, 1st edn. Informa Healthcare, UK, pp 16–17
Poole CA, Flint MH, Beaumont BW (1985) Analysis of the morphology and function of primary cilia in connective tissues: a cellular cybernetic probe? Cell Motil 5:175–193
Reese J, Brown N, Das SK, Dey SK (1998) Expression of neu differantation factor during the periimplantation period in the mouse uterus. Biol Reprod 58:719–727
Ryan JN, Woessner JF (1972) Oestradiol inhibits collagen breakdown in the involuting rat uterus. Biochem J 127:705–713
Ryvnyak VV, Gudumak VS, Rybakova MA, Grumeza OF, Pelin AV (1999) Extra-and intracellular collagen resorption by smooth muscle cells in postpartum uterine involution. Bull Exp Biol Med 127:96–98
Sağsöz H, Ketani MA (2010) The role of estrogen receptors, erbB receptors, vascular endothelial growth factor and its receptors, and vascular endothelial growth inhibitor in the development of the rat mammary gland. Growth Factors 28:379–393
Sağsöz H, Ketani MA, Saruhan BG (2012) Expression of the erbB/HER receptor family in the bovine uterus during the sexual cycle and the relation of this family to serum sex steroids. Biotech Histochem 87:105–116
Sağsöz H, Liman N, Güney Saruhan B, Küçükaslan I (2014) The expression of epidermal growth factor receptors and their ligands (epidermal growth factor, neuregulin, amphiregulin) in the bitch uterus during the estrus cyle. Anim Reprod Sci 147:161–179. doi:10.1016/j.anireprosci/2014.04.005
Salamonsen LA (2003) Tissue injury and repair in the female human reproductive tract. Reproduction 125:301–311
Salgado RM, Favaro RR, Zorn TMT (2011) Modulation of small leucine-rich proteoglycans (SLRPs) expression in the mouse uterus by estradiol and progesterone. Reprod Biol Endocrinol 9:22
Schneider MR, Wolf E (2008) The epidermal growth factor receptor and its ligands in female reproduction: insights from rodent models. Cytokine Growth Factor Rev 19:173–181
Shiraga M, Komatsu N, Teshigawara K et al (2000) Epidermal growth factor stimulates proliferation of mouse uterine epithelial cells in primary culture. Zool Sci 17:661–666
Shoyab M, McDonald VL, Bradley JG, Todaro GJ (1988) Amphiregulin: a bifunctional growth-modulating glycoprotein produced by the phorbol 12-myristate 13-acetate-treated human breast adenocarcinoma cell line MCF-7. Proc Natl Acad Sci U S A 85:6528–6532
Shynlova O, Oldenhof A, Dorogin A et al (2006) Myometrial apoptosis: activation of the caspase cascade in the pregnant rat myometrium at midgestation. Biol Reprod 74:839–849
Simpson KR, Creehan PA (2008) Perinatal nursing. Lippincott Williams & Wilkins, Philadelphia, p 485
Siziopikou KP, Khan S (2005) Correlation of HER2 gene amplification with expression of the apoptosis-suppressing genes bcl-2 and bcl-x-L in ductal carcinoma in situ of the breast. Appl Immunohistochem Mol Morphol 13:14–18
Slowey MJ, Verhage HG, Fazleabas AT (1994) Epidermal growth factor, transforming growth factor-α, and epidermal growth factor receptor localization in the baboon (Papio anubis) uterus during the menstrual cycle and early pregnancy. J Soc Gynecol Investig 1:277–284
Smith K, Lejeune S, Harris AH, Rees MCP (1991) Epidermal growth factor receptor in human uterine tissues. Hum Reprod 6:619–622
Sorkin A, Goh LK (2008) Endocytosis and intracellular trafficking of ErbBs. Exp Cell Res 314:3093–3106
Srinivasan R, Poulsom R, Hurst HC, Gullick WJ (1998) Expression of the c-erbB-4/HER4 protein and mRNA in normal human fetal and adult tissues and in a survey of nine solid tumour types. J Pathol 185:236–245
Srinivasan R, Benton E, McCormick F, Thomas H, Gullick WJ (1999) Expresssion of the c-erbB3/HER-3 and c-erbB4/HER-4 growth factor receptors and their ligands, neuregulin-1α, neuregulin-1β, and betacellulin, in normal endometrium and endometrial cancer. Clin Cancer Res 5:2877–2883
Stancel GM, Gardner RM, Kirkland JL et al (1987) Interactions between estrogen and EGF in uterine growth and function. Adv Exp Med Biol 230:99–118
Stern DF (2003) ErbBs in mammary development. Exp Cell Res 284:89–98
Tachi S, Tachi C, Linder HR (1969) Cilia-bearing stromal cells in the rat uterus. J Anat 104:295–308
Tamada H, Yoh C, Inaba T, Takano H, Kawate N, Sawada T (2000) Epidermal growth factor (EGF) in the goat uterus: immunohistochemical localization of EGF and EGF receptor and effectof EGF on uterine activity in vivo. Therigenology 54:159–169
Tamada H, Tsubutani D, Kawate N et al (2002) Detection of transforming growth factor-α and epidermal growth factor receptor mRNA and immunohistochemical localization of their proteins in the ovine uterus during the early implantation period. Histochem J 34:383–390
Tamada H, Tominaga M, Kida K et al (2005) Detection of transforming growth factor-a andepidermal growth factor receptor mRNA and immunohistochemical localization of the corresponding proteins in the canine uterus duringthe estrous cycle. Histol Histopathol 20:817–824
Tsark EC, Adamson ED, Withers GE, Wiley LM (1997) Expression and function of amphiregulin during murine preimplantation development. Mol Reprod Dev 47:271–283
Wang YN, Hung MC (2012) Nuclear functions and subcellular trafficking mechanisms of the epidermal growth factor receptor family. Cell Biosci 2:13
Wang D, Fujii S, Konishi I, Nanbu Y, Iwai T, Nonogaki H (1992) Expression of c-erbB2 protein and epidermal growth factor receptor in normal tissues of the female genital tract and in tha placenta. Virchows Arch A Pathol Anat Histopathol 420:385–393
Wang Y, Pennock S, Chen SX, Wang Z (2002) Endosomal signaling of epidermal growth factor receptor stimulates signal transduction pathways leading to cell survival. Mol Cell Biol 22:7279–7290
Warbrick JG (1955) Postpartum changes in the uterus of rat. J Embryol Exp Morphol 3:256–264
Werb Z (1990) Expression of EGF and TGF-α genes in early mammalian development. Mol Reprod Dev 27:10–15
Wollenhaupt K, Tiemann U, Einspanier R, Schneider F, Kanitz W, Brüssow K-P (1997) Characterization of the epidermal growth factor receptor in pig oviduct and endometrium. J Reprod Fertil 111:173–181
Wollenhaupt K, Welter H, Einspanier R, Manabe N, Brüssow K-P (2004) Expression of epidermal growth factor receptor (EGF-R), vascular endothelial growth factor receptor (VEGF-R) and fibroblast growth factor receptor (FGF-R) systems in porcine oviduct and endometrium during the time of implantation. J Reprod Dev 50:269–278
Wong T, McGrath JA, Navsaria H (2007) The role of fibroblasts in tissue engineering and regeneration. Br J Dermatol 156:1149–1155
Yamanaka Y, Hayashi K, Komurasaki T, Morimoto S, Ogihara T, Sobue K (2001) EGF family ligand-dependent phenotypic modulation of smooth muscle cells through EGF receptor. Biochem Biophys Res Commun 281:373–377
Yarden Y, Sliwkowski MX (2001) Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2:127–137
Yeh J, Rein M, Nowak R (1991) Presence of messenger ribonucleic acid for epidermal growth factor (EGF) and EGF receptor demonstrable in monolayer cell cultures of myometria and leiomyomata. Fertil Steril 56:997–1000
Yu W-H, Woessner JF, McNeish JD, Starnenkovic I (2002) CD44 anchors the assembly of matrilysin/MMP-7 with heparin-binding epidermal growth factor precursor and ErbB4 and regulates female reproductive organ remodeling. Genes Dev 16:307–323
Yue Z-P, Yang Z-M, Li S-J, Wang H-B, Haroer MJK (2000) Epidermal growth factor family in rhesus monkey uterus during the menstrual cycle and early pregnancy. Mol Reprod Dev 55:164–174
Zhang Z, Krause M, Davis DL (1992) Epidermal growth factor receptors in porcine endometrium: binding characteristics and the regulation of prostaglandin E and F2 alpha production. Biol Reprod 46:932–936
Acknowledgments
This study was financially supported by Scientific Research Council of Erciyes University (Project No: TSA-12-3865).
Conflict of interest
The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Alan, E., Liman, N. & Sağsöz, H. The profile of the epidermal growth factor system in rat endometrium during postpartum involution period. Vet Res Commun 39, 115–135 (2015). https://doi.org/10.1007/s11259-015-9633-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11259-015-9633-6