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Gender- and region-specific variations of estrogen receptor α and β expression in the growth plate of spine and limb during development and adulthood

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Abstract

Although estrogen action is indispensable for normal bone growth in both genders, the roles of estrogen receptors (ERs) in mediating bone growth are not fully understood. The effects of ER inactivation on bone growth are sex and age dependent, and may differ between the axial and appendicular regions. In this study, the spatial and temporal expression of ERα and β in the tibial and spinal growth plates of the female and male rats during postnatal development was examined to explore the possible mechanisms. The level of mRNA was examined and compared with quantitative real-time PCR. The spatial location was determined by immunohistochemical analysis. The 1-, 4-, 7-, 12- and 16-week age stages correspond to early life, puberty and early adulthood after puberty, respectively. Gender- and region-specific differences in ERα and β expression were shown in the growth plates. Mainly nuclear staining of ERα and β immunoreactivity was demonstrated in the spinal and tibial growth plate chondrocytes for both genders. Moreover, our study indicated significant effect of gender on temporal ERα and β expression and of region on temporal ERα/ERβ expression ratio. However, spatial differences of region-related ERα and β expression were not observed. Gender-related spatial changes were detected only at 16 weeks of both spine and limb growth plates. ERα and β immunoreactivity was detected in the resting, proliferative and prehypertrophic chondrocytes in the early life stage and during puberty. After puberty, ERα expression was mainly located in the late proliferative and hypertrophic chondrocytes in female, whereas the expression still extended from the resting to hypertrophic chondrocytes in males. Gender- and region-specific expression patterns of ERα and β gene might be one possible reason for differences in sex- and region-related body growth phenotypes. Gender, age and region differences should be taken into consideration when the roles of ERs in the growth plate are investigated.

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References

  • Bass S, Delmas PD, Pearce G, Hendrich E, Tabensky A, Seeman E (1999) The differing tempo of growth in bone size, mass, and density in girls is region-specific. J Clin Invest 104:795–804

    Article  PubMed  CAS  Google Scholar 

  • Ben-Hur H, Mor G, Blickstein I, Likhman I, Kohen F, Dgani R, Insler V, Yaffe P, Ornoy A (1993) Localization of estrogen receptors in long bones and vertebrae of human fetuses. Calcif Tissue Int 53:91–96

    Article  PubMed  CAS  Google Scholar 

  • Bilezikian JP, Morishima A, Bell J, Grumbach MM (1998) Increased bone mass as a result of estrogen therapy in a man with aromatase deficiency. N Engl J Med 339:599–603

    Article  PubMed  CAS  Google Scholar 

  • Bord S, Horner A, Beavan S, Compston J (2001) Estrogen receptors alpha and beta are differentially expressed in developing human bone. J Clin Endocrinol Metab 86:2309–2314

    Article  PubMed  CAS  Google Scholar 

  • Bradney M, Karlsson MK, Duan Y, Stuckey S, Bass S, Seeman E (2000) Heterogeneity in the growth of the axial and appendicular skeleton in boys: implications for the pathogenesis of bone fragility in men. J Bone Miner Res 15:1871–1878

    Article  PubMed  CAS  Google Scholar 

  • Braidman IP, Davenport LK, Carter DH, Selby PL, Mawer EB, Freemont AJ (1995) Preliminary in situ identification of estrogen target cells in bone. J Bone Miner Res 10:74–80

    Article  PubMed  CAS  Google Scholar 

  • Braidman IP, Hainey L, Batra G, Selby PB, Saunders PT, Hoyland JA (2001) Localization of estrogen receptor protein expression in adult human bone. J Bone Miner Res 16:214–220

    Article  PubMed  CAS  Google Scholar 

  • Buie HR, Moore CP, Boyd SK (2008) Postpubertal architectural developmental patterns differ between the L3 vertebra and proximal tibia in three inbred strains of mice. J Bone Miner Res 23:2048–2059

    Article  PubMed  Google Scholar 

  • Byers S, Moore AJ, Byard RW, Fazzalari NL (2000) Quantitative histomorphometric analysis of the human growth plate from birth to adolescence. Bone 27:495–501

    Article  PubMed  CAS  Google Scholar 

  • Camper L, Holmvall K, Wängnerud C, Aszódi A, Lundgren-Akerlund E (2001) Distribution of the collagen-binding integrin alpha10beta1 during mouse development. Cell Tissue Res 306:107–116

    Google Scholar 

  • Carani C, Qin K, Simoni M, Faustini-Fustini M, Serpente S, Boyd J, Korach KS, Simpson ER (1997) Effect of testosterone and estradiol in a man with aromatase deficiency. N Engl J Med 337:91–95

    Article  PubMed  CAS  Google Scholar 

  • Chagin AS, Sävendahl L (2009) Genes of importance in the hormonal regulation of growth plate cartilage. Horm Res 71(Suppl 2):41–47

    Article  PubMed  CAS  Google Scholar 

  • Chagin AS, Lindberg MK, Andersson N, Moverare S, Gustafsson JA, Sävendahl L, Ohlsson C (2004) Estrogen receptor-β inhibits skeletal growth and has the capacity to mediate growth plate fusion in female mice. J Bone Miner Res 19:72–77

    Article  PubMed  CAS  Google Scholar 

  • Couse JF, Korach KS (1999) Estrogen receptor null mice: what have we learned and where will they lead us? Endocr Rev 20:358–417

    Article  PubMed  CAS  Google Scholar 

  • Egerbacher M, Helmreich M, Rossmanith W, Haeusler G (2002) Estrogen receptor-α and estrogen receptor-β are present in the human growth plate in childhood and adolescence, in identical distribution. Horm Res 58:99–103

    Article  PubMed  CAS  Google Scholar 

  • Fukunaga T, Yamashiro T, Oya S, Takeshita N, Takigawa M, Takano-Yamamoto T (2003) Connective tissue growth factor mRNA expression pattern in cartilages is associated with their type I collagen expression. Bone 33:911–918

    Google Scholar 

  • Gevers EF, van der Eerden BC, Karperien M, Raap AK, Robinson IC, Wit JM (2002) Localization and regulation of the growth hormone receptor and growth hormone-binding protein in the rat growth plate. J Bone Miner Res 17:1408–1419

    Article  PubMed  CAS  Google Scholar 

  • Giustina A, Mazziotti G, Canalis E (2008) Growth hormone, insulin-like growth factors, and the skeleton. Endocr Rev 29:535–559

    Article  PubMed  CAS  Google Scholar 

  • Glatt V, Canalis E, Stadmeyer L, Bouxsein ML (2007) Age-related changes in trabecular architecture differ in female and male C57BL/6J mice. J Bone Miner Res 22:1197–1207

    Article  PubMed  Google Scholar 

  • Grumbach MM (2000) Estrogen, bone, growth and sex: a sea change in conventional wisdom. J Pediatr Endocrinol Metab 13(Suppl 6):1439–1455

    PubMed  Google Scholar 

  • Hosoya A, Hoshi K, Sahara N, Ninomiya T, Akahane S, Kawamoto T, Ozawa H (2005) Effects of fixation and decalcification on the immunohistochemical localization of bone matrix proteins in fresh-frozen bone sections. Histochem Cell Biol 123:639–646

    Article  PubMed  CAS  Google Scholar 

  • Hughes PC, Tanner JM (1970) The assessment of skeletal maturity in the growing rat. J Anat 106:371–402

    PubMed  CAS  Google Scholar 

  • Kaludjerovic J, Ward WE (2008) Diethylstilbesterol has gender-specific effects on weight gain and bone development in mice. J Toxicol Environ Health A 71:1032–1042

    Article  PubMed  CAS  Google Scholar 

  • Kennedy J, Baris C, Hoyland JA, Selby PL, Freemont AJ, Braidman IP (1999) Immunofluorescent localization of estrogen receptor-α in growth plates of rabbits, but not in rats, at sexual maturity. Bone 24:9–16

    Article  PubMed  CAS  Google Scholar 

  • Khosla S, Riggs BL, Atkinson EJ, Oberg AL, McDaniel LJ, Holets M, Peterson JM, Melton LJ III (2006) Effects of sex and age on bone microstructure at the ultradistal radius: a population-based noninvasive in vivo assessment. J Bone Miner Res 21:124–131

    Article  PubMed  Google Scholar 

  • Kronenberg HM (2003) Developmental regulation of the growth plate. Nature 423:332–336

    Article  PubMed  CAS  Google Scholar 

  • Kusec V, Virdi AS, Prince R, Triffitt JT (1998) Localization of estrogen receptor- in human and rabbit skeletal tissues. J Clin Endocrinol Metab 83:2421–2428

    Article  PubMed  CAS  Google Scholar 

  • Laue L, Kenigsberg D, Pescovitz OH, Hench KD, Barnes KM, Loriaux DL, Cutler GB Jr (1989) Treatment of familial male precocious puberty with spironolactone and testolactone. N Engl J Med 320:496–502

    Article  PubMed  CAS  Google Scholar 

  • Lebovitz HE, Eisenbarth GS (1975) Hormonal regulation of cartilage growth and metabolism. Vitam Horm 33:575–648

    Article  PubMed  CAS  Google Scholar 

  • Lee PA, Witchel SF (1997) The influence of estrogen on growth. Curr Opin Pediatr 9:431–436

    Article  PubMed  CAS  Google Scholar 

  • Li H, Jiang LS, Dai LY (2010) High glucose potentiates collagen synthesis and bone morphogenetic protein-2-induced early osteoblast gene expression in rat spinal ligament cells. Endocrinology 151:63–74

    Article  PubMed  CAS  Google Scholar 

  • Li XF, Yan J, Jiang LS, Dai LY (2011) Age-related variations of leptin receptor expression in the growth plate of spine and limb: gender- and region-specific changes. Histochem Cell Biol 135:487–497

    Google Scholar 

  • Lindberg MK, Alatalo SL, Halleen JM, Mohan S, Gustafsson JA, Ohlsson C (2001) Estrogen receptor specificity in the regulation of the skeleton in female mice. J Endocrinol 171:229–236

    Article  PubMed  CAS  Google Scholar 

  • Maffei L, Murata Y, Rochira V, Tubert G, Aranda C, Vazquez M, Clyne CD, Davis S, Simpson ER, Carani C (2004) Dysmetabolic syndrome in a man with a novel mutation of the aromatase gene: effects of testosterone, alendronate, and estradiol treatment. J Clin Endocrinol Metab 89:61–70

    Article  PubMed  CAS  Google Scholar 

  • Matthews JB (1982) Influence of decalcification on immunohistochemical staining of formalin-fixed paraffin-embedded tissue. J Clin Pathol 35:1392–1394

    Article  PubMed  CAS  Google Scholar 

  • Mitani H, Takahashi I, Onodera K, Bae JW, Sato T, Takahashi N, Sasano Y, Igarashi K, Mitani H (2006) Comparison of age-dependent expression of aggrecan and ADAMTSs in mandibular condylar cartilage, tibial growth plate, and articular cartilage in rats. Histochem Cell Biol 126:371–380

    Article  PubMed  CAS  Google Scholar 

  • Morishima A, Grumbach MM, Simpson ER, Fisher C, Qin K (1995) Aromatase deficiency in male and female siblings caused by a novel mutation and the physiological role of estrogens. J Clin Endocrinol Metab 80:3689–3698

    Article  PubMed  CAS  Google Scholar 

  • Mundlos S, Zabel B (1994) Developmental expression of human cartilage matrix protein. Dev Dyn 199:241–252

    Google Scholar 

  • Neves JD, Omar NF, Narvaes EA, Gomes JR, Novaes PD (2011) Influence of different decalcifying agents on EGF and EGFR immunostaining. Acta Histochem 113:484–488

    Article  CAS  Google Scholar 

  • Nilsson LO, Boman A, Sävendahl L, Grigelioniene G, Ohlsson C, Ritzén EM, Wroblewski J (1999) Demonstration of estrogen receptor-β immunoreactivity in human growth plate cartilage. J Clin Endocrinol Metab 84:370–373

    Article  PubMed  CAS  Google Scholar 

  • Nilsson O, Abad V, Chrysis D, Ritzén EM, Sävendahl L, Baron J (2002) Estrogen receptor-α and -β are expressed throughout postnatal development in the rat and rabbit growth plate. J Endocrinol 173:407–414

    Article  PubMed  CAS  Google Scholar 

  • Nilsson O, Chrysis D, Pajulo O, Boman A, Holst M, Rubinstein J, Martin Ritzén E, Sävendahl L (2003) Localization of estrogen receptors-α and -β and androgen receptor in the human growth plate at different pubertal stages. J Endocrinol 177:319–326

    Article  PubMed  CAS  Google Scholar 

  • Nilsson O, Marino R, De Luca F, Phillip M, Baron J (2005) Endocrine regulation of the growth plate. Horm Res 64:157–165

    Article  PubMed  CAS  Google Scholar 

  • Ornoy A, Giron S, Aner R, Goldstein M, Boyan BD et al (1994) Gender dependent effects of testosterone and 17 β-estradiol on bone growth and modelling in young mice. Bone Miner 24:43–58

    Article  PubMed  CAS  Google Scholar 

  • Parikka V, Peng Z, Hentunen T, Risteli J, Elo T, Väänänen HK, Härkönen P (2005) Estrogen responsiveness of bone formation in vitro and altered bone phenotype in aged estrogen receptor-α deficient male and female mice. Eur J Endocrinol 152:301–314

    Article  PubMed  CAS  Google Scholar 

  • Sampson HW, Dearman AC, Akintola AD, Zimmer WE, Parrish AR (2007) Immunohistochemical localization of cadherin and catenin adhesion molecules in the murine growth plate. J Histochem Cytochem 55:845–852

    Article  PubMed  CAS  Google Scholar 

  • Seeman E (2001) Unresolved issues in osteoporosis in men. Rev Endocr Metab Disord 2:45–64

    Article  PubMed  CAS  Google Scholar 

  • Sims NA, Dupont S, Krust A, Clement-Lacroix P, Minet D, Resche-Rigon M, Gaillard-Kelly M, Baron R (2002) Deletion of estrogen receptors reveals a regulatory role for estrogen receptors-β in bone remodeling in females but not in males. Bone 30:18–25

    Article  PubMed  CAS  Google Scholar 

  • Smith EP, Boyd J, Frank GR, Takahashi H, Cohen RM, Specker B, Williams TC, Lubahn DB, Korach KS (1994) Estrogen resistance caused by a mutation in the estrogen-receptor gene in a man. N Engl J Med 331:1056–1061

    Article  PubMed  CAS  Google Scholar 

  • Tozum TF, Oppenlander ME, Koh-Paige AJ, Robins DM, McCauley LK (2004) Effects of sex steroid receptor specificity in the regulation of skeletal metabolism. Calcif Tissue Int 75:60–70

    Article  PubMed  CAS  Google Scholar 

  • Tsunenari T, Tsutsumi M, Ohno K, Yamamoto Y, Kawakatsu M, Shimogaki K, Negishi H, Sugimoto T, Fukase M, Fujita T (1993) Age- and gender-related changes in body composition in Japanese subjects. J Bone Miner Res 8:397–402

    Article  PubMed  CAS  Google Scholar 

  • van der Eerden BC, Karperien M, Gevers EF, Löwik CW, Wit JM (2000) Expression of Indian hedgehog, parathyroid hormone-related protein, and their receptors in the postnatal growth plate of the rat: evidence for a locally acting growth restraining feedback loop after birth. J Bone Miner Res 15:1045–1055

    Article  PubMed  Google Scholar 

  • van der Eerden BCJ, Karperien M, Wit JM (2001) The estrogen receptor in the growth plate: implications for pubertal growth. J Pediatr Endocrinol Metab 14:1527–1533

    PubMed  Google Scholar 

  • van der Eerden BCJ, Gevers EF, Lowik CW, Karperien M, Wit JM (2002) Expression of estrogen receptor and in the epiphyseal plate of the rat. Bone 30:478–485

    Article  PubMed  Google Scholar 

  • Vidal O, Lindberg M, Sävendahl L, Lubahn DB, Ritzen EM, Gustafsson JA, Ohlsson C (1999) Disproportional body growth in female estrogen receptor-α-inactivated mice. Biochem Biophys Res Commun 265:569–571

    Article  PubMed  CAS  Google Scholar 

  • Vidal O, Lindberg MK, Hollberg K, Baylink DJ, Andersson G, Lubahn DB, Mohan S, Gustafsson JA, Ohlsson C (2000) Estrogen receptor specificity in the regulation of skeletal growth and maturation in male mice. Proc Natl Acad Sci USA 97:5474–5479

    Article  PubMed  CAS  Google Scholar 

  • Windahl SH, Vidal O, Andersson G, Gustafsson JA, Ohlsson C (1999) Increased cortical bone mineral content but unchanged trabecular bone mineral density in female ERβ (−/−) mice. J Clin Invest 104:895–901

    Article  PubMed  CAS  Google Scholar 

  • Windahl SH, Hollberg K, Vidal O, Gustafsson JA, Ohlsson C, Andersson G (2001) Female estrogen receptor β−/− mice are partially protected against age-related trabecular bone loss. J Bone Miner Res 16:1388–1398

    Article  PubMed  CAS  Google Scholar 

  • Yilmaz D, Ersoy B, Bilgin E, Gümüşer G, Onur E, Pinar ED (2005) Bone mineral density in girls and boys at different pubertal stages: relation with gonadal steroids, bone formation markers, and growth parameters. J Bone Miner Metab 23:476–482

    Article  PubMed  CAS  Google Scholar 

  • Yuen KW, Kwok TC, Qin L, Leung JC, Chan DC, Kwok AW, Woo J, Leung PC (2010) Characteristics of age-related changes in bone compared between male and female reference Chinese populations in Hong Kong: a pQCT study. J Bone Miner Metab 28:672–681

    Article  PubMed  Google Scholar 

  • Zachmann M, Prader A, Sobel EH, Crigler JF Jr, Ritzén EM, Atarés M, Ferrandez A (1986) Pubertal growth in patients with androgen insensitivity: indirect evidence for the importance of estrogens in pubertal growth of girls. J Pediatr 108:694–697

    Article  PubMed  CAS  Google Scholar 

  • Zhang XZ, Kalu DN, Erbas B, Hopper JL, Seeman E (1999) The effects of gonadectomy on bone size, mass, and volumetric density in growing rats are gender-, site-, and growth hormone-specific. J Bone Miner Res 14:802–809

    Article  PubMed  CAS  Google Scholar 

  • Zhao CQ, Liu D, Li H, Jiang LS, Dai LY (2008) Expression of leptin and its functional receptor on disc cells: contribution to cell proliferation. Spine 33:E858–E864

    Article  PubMed  Google Scholar 

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (30901508, U1032001).

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  1. Department of Orthopedic Surgery, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, 1665 Kongjiang Road, 200092, Shanghai, China

    Xin-Feng Li, Shan-Jin Wang, Lei-Sheng Jiang & Li-Yang Dai

  2. Department of Orthopedic Surgery, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

    Xin-Feng Li

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  1. Xin-Feng Li
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Correspondence to Li-Yang Dai.

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X. -F. Li and S. -J. Wang contributed equally to this paper.

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Li, XF., Wang, SJ., Jiang, LS. et al. Gender- and region-specific variations of estrogen receptor α and β expression in the growth plate of spine and limb during development and adulthood. Histochem Cell Biol 137, 79–95 (2012). https://doi.org/10.1007/s00418-011-0877-0

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