GPER-ERK通路在DES影响小鼠睾丸引带发育中的作用

doi:10.3969/j.issn.1004-616x.2015.02.008

Abstract

Abstract:

OBJECTIVE:To explore the presence of G protein-coupled estrogen receptor (GPER) in the Kunming mouse gubernaculum testis. To explore the rapid effects of diethylstilbestrol (DES) on cultured mouse gubernaculum testis cells via ERK signaling pathway. METHODS:The expression of GPER was assessed by immunohistochemical staining with the development of mouse gubernaculums testis (GD17, GD19, PD0, PD3, PD7, PD14 and PD21). Subcellular localization of GPER was identified by Immunoelectron Microscopy. Cells subcultured from mouse gubernaculum testis were divided into three groups randomly, that is, control group(DES 1.5×10^-8 mol/L), ICI 182780-DES group (pretreated with 1 nmol/L estrogen receptor inhibitor ICI 182780 90 min), and G15-DES group (pretreated with 1 nmol/L GPER inhibitor G15 90 min). p-ERK1/2 was detected in the cells with 1.5×10^-8 mol/L DES in different groups by Western blot. RESULTS:The immunohistochemical staining showed that GPER was expressed in the development of gubernaculums testis, mainly in the unconsolidated mesenchymal tissue of inner gubernaculum testis, and GPER staining was strong both on the membrane and in the cytoplasm. The subcellular localization of GPER protein was mainly in the mesh-like network of cytoplasm, the rough endoplasmic reticulum. G15 could reverse the DES-induced inhibitory effect of p-ERK1/2, but ICI182780 didn't have the same effect. CONCLUSION:GPER was expressed in membrane structure of gubernaculums testis cells in different stages of development. The nongenetic effects of DES on gubernaculums testis cells were likely to be partly mediated by GPER. Therefore, our data provide new insight into the role of EEs in the etiology of male reproductive system and will help develop better approaches for the prevention and therapy of male reproductive malformation.

Key words: gubernaculum testis, G protein-coupled estrogen receptor, environment estrogens, diethylstilbestrol

CLC Number:

R994.6

ZHANG Xuan, KE Song, LIU Liang, CHEN Kaihong, MA Lian, JIANG Xuewu. Effects of DES on mouse gubernaculum testis development via GPER-ERK signaling pathway[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(2): 116-120,124.

References

[1] Schultz MM, Minarik TA, Martinovic-Weigelt D, et al. Environmental estrogens in an urban aquatic ecosystem:II. Biological effects[J]. Environ Int, 2013, 61:138-149.
[2] Skakkebaek NE, Rajpert D, Meyts E, et al. Testicular dysgenesis syndrome:an increasingly common developmental disorder with environmental aspects[J]. Hum Reprod, 2001, 16(5):972-978.
[3] Toppari J, Virtanen HE, Main KM, et al. Cryptorchidism and hypospadias as a sign of testicular dysgenesis syndrome (TDS): environmental connection[J]. Birth Defects Res A Clin Mol Teratol, 2010, 88(10):910-919.
[4] Fernandez MF1, Duran I, Olea N, et al. Semen quality and reproductive hormone levels in men from Southern Spain[J]. Int J Androl, 2012, 35(1):1-10.
[5] 蒋学武, 邓汪东, 胡显良, 等. 外源性雌激素对小鼠睾丸引带收缩和增殖活性的影响[J]. 中华小儿外科杂志, 2005, 26(9):443-446.
[6] Barthold JS, Robbins A, Wang Y, et al. Cryptorchidism in the orl rat is associated with muscle patterning defects in the fetal gubernaculum and alteredhormonal signaling[J]. Biol Reprod, 2014, 91(2):41.
[7] 蒋学武, 陈中献, 陈绍基, 等. 先天性尿道下裂阴茎多种性激素受体的比较研究[J]. 中国男科学杂志, 2000, 14(1): 16-18.
[8] 胡廷泽, 蒋学武, 刘文英, 等. 青春期前小阴茎患儿的临床和内分泌观察[J]. 中华小儿外科杂志, 1997, 18(1):32-34.
[9] 王光义, 陈绍基, 蒋学武, 等. 尿道下裂合并阴茎发育不良的内泌改变及治疗[J]. 中华小儿外科杂志, 1995, 16(1): 23-25.
[10] 蒋学武, 陈绍基, 王光义, 等. 先天性尿道下裂阴茎雄激素受体的研究[J]. 中华小儿外科杂志, 1995, 16(3):141-142.
[11] 邓汪东, 蒋学武, 李建宏, 等. 己烯雌酚致胚胎期睾丸引带形态结构发育异常的研究[J]. 癌变·畸变·突变, 2005, 17(2):93-96.
[12] Jiang XW, Li JH, Huang TH, et al. Effect of prenatal exposure to diethylstilbestrol on gubernacular development in fetal male mice[J]. Asian J Androl, 2004, 6(4):325-329.
[13] 张镟, 蒋学武, 李建宏, 等. 小鼠睾丸引带细胞的培养及己烯雌酚对培养睾丸引带细胞的影响[J]. 中华男科学杂志, 2009, 15(10):872-875.
[14] Zhang Xuan, Li Jian-hong, Ma Lian, et al. Diethylstilbestrol impairs the morphology and function of mouse gubernaculum testis in culture[J]. Cell Biol Toxicol, 2012, 28(6):397-407.
[15] Zhang X, Li JH, Duan SX, et al. G protein-coupled estrogen receptor-protein kinase A-ERK-CREB signaling pathway is involved in the regulation of mouse gubernaculum testis cells by diethylstilbestrol[J]. Arch Environ Contam Toxicol, 2014, 67(1):97-103.
[16] Roger C, Lambard S, Bouskine A, et al. Estrogen-induced growth inhibition of human seminoma cells expressing estrogen receptor beta and aromatase[J]. J Mol Endocrinol, 2005, 35(1):191-199.
[17] Bouskine A, Nebout M, Mograbi B, et al. Estrogens promote human testicular germ cell cancer through a membrane-mediated activation of extracellular regulated kinase and protein kinase A[J]. Endocrinology, 2008, 149(2):565-573.
[18] Revankar CM, Cimino DF, Sklar LA, et al. A transmembrane intracellular estrogen receptor mediates rapid cell signaling[J]. Science, 2005, 307(5715):1625-1630.
[19] Olde B, Leeb-Lundberg LM. GPR30/GPER1:searching for a role in estrogen physiology[J]. Trends Endocrinol Metab, 2009, 20(8):409-416.
[20] Prossnitz ER, Maggiolini M. Mechanisms of estrogen signaling and gene expression via GPR30[J]. Mol Cell Endocrinol, 2009, 308(1/2):32-38.
[21] Thorup J, McLachlan R, Cortes D, et al. What is new in cryptorchidism and hypospadias-a critical review on the testicular dysgenesis hypothesis[J]. J Pediatr Surg, 2010, 45(10):2074-2086.
[22] Nagraj S, Seah GJ, Farmer PJ, et al. The development and anatomy of the gubernaculum in Hoxa11 knockout mice[J]. J Pediatr Surg, 2011, 46(2):387-392.
[23] Kong RC, Shilling PJ, Lobb DK, et al. Membrane receptors: structure and function of the relaxin family peptide receptors[J]. Mol Cell Endocrinol, 2010, 320(1/2):1-15.
[24] Walker CS, Conner AC, Poyner DR, et al. Regulation of signal transduction by calcitonin gene-related peptide receptors [J]. Trends Pharmacol Sci, 2010, 31(10):476-483.
[25] Viñas R, Jeng YJ, Watson CS. Non-genomic effects of xenoestrogen mixtures[J]. Int J Environ Res Public Health, 2012, 9(8):2694-714.
[26] Maggiolini M, Picard D. The unfolding stories of GPR30, a new membrane-bound estrogen receptor[J]. J Endocrinol, 2010, 204(2):105-114.
[27] Filardo EJ. Estrogen-induced activation of Erk-1 and Erk-2 requires the G protein-coupled receptor homolog, GPR30, and occurs via trans-activation of the epidermal growth factor receptor through release of HB-EGF[J]. Mol Endocrinol, 2000, 14:1649-1660.

Effects of DES on mouse gubernaculum testis development via GPER-ERK signaling pathway

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 2

Recommended Articles

Metrics

Comments

[1]	ZHANG Yu_mei, SUN Xia, YIN Jun, PAN Jin_chun. Building Lung Tumor Model of New Born Mice Induced by Diethylstilbestrol [J]. , 2006, 18(6): 462-464.
[2]	DENG Wang-dong;JIANG Xue-wu;LI Jian-hong;LAI Ya-man;CHEN Zhong-xian; HUANG Tian-hua. The Effect of Diethylstilbestrol on Gubernaculum Testis Development in Fetal Mice [J]. , 2005, 17(2): 93-96.