肾上腺素能受体β2和胆碱能受体M1在小鼠睾丸的表达定位及功能研究

doi:10.11843/j.issn.0366-6964.2017.06.007

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (6): 1028-1034.doi: 10.11843/j.issn.0366-6964.2017.06.007

肾上腺素能受体β2和胆碱能受体M1在小鼠睾丸的表达定位及功能研究

霍书英*, 李玉荣, 武现军

河北农业大学动物医学院, 保定 071001

收稿日期:2016-12-01 出版日期:2017-06-23 发布日期:2017-06-23
通讯作者: 霍书英,Tel:0312-7528344,E-mail:huoshuying@163.com
作者简介:霍书英(1974-),女,河北灵寿人,副教授,博士,主要从事动物生殖内分泌研究
基金资助:
河北省教育厅科学研究项目（Z2013021）；国家自然科学基金（31172094）

Study on the Expression and Functions of Adrenergic Receptor β2 and Muscarinic Acetylcholine Receptor M1 in Mouse Testis

HUO Shu-ying*, LI Yu-rong, WU Xian-jun

College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China

Received:2016-12-01 Online:2017-06-23 Published:2017-06-23

摘要/Abstract

摘要：

旨在研究肾上腺素能受体β2和胆碱能受体M1在小鼠睾丸组织的表达定位及其生理功能。采用RT-PCR方法检测β2受体和M1受体mRNA在小鼠睾丸不同发育时期的表达变化；免疫组织化学方法研究β2受体和M1受体在小鼠睾丸的表达定位；通过体外细胞培养方法研究睾丸间质细胞β2受体和M1受体的生理功能。结果表明：β2受体和M1受体mRNA在小鼠睾丸不同发育时期均显著表达；β2受体和M1受体主要在睾丸间质细胞表达； 10^-5和10^-6mol·L^-1经递质 NE和Ach均能显著促进小鼠睾丸间质瘤细胞MLTC-1的增殖（P<0.05），β2受体阻断剂布托沙明（Buto）和M1受体阻断剂哌吡氮平（Pire）均能阻断NE和Ach的促增殖作用（P<0.05）；NE和Ach均能明显抑制MLTC-1细胞3β-HSD mRNA的表达，其阻断剂Buto和Pire均能阻断其作用效果；NE可明显促进雌激素受体α（ERα）mRNA的表达，而Ach可明显促进ERβ的表达，但其阻断剂Buto和Pire并不能阻断NE 和Ach促进ERα和ERβ mRNA表达的作用效果。综上表明，NE和Ach调节睾丸间质细胞的数量和雄激素的合成，主要是通过分布于睾丸间质细胞的功能受体β2受体和M1受体介导的，但NE和Ach调节雌激素受体ERα和ERβ的表达并不受β2受体和M1受体的介导。

Abstract:

The experiment was conducted to study the expression and functions of adrenergic receptor β2 (β2AR) and muscarinic acetylcholine receptor M1 (M1R) in mouse testis. The expression of β2AR and M1R mRNA in mouse testis was analysed by method of RT-PCR at different developmental stages. Expression and localization of β2AR and M1R in mouse testis were examined by immunohistochemistry. The mouse Leydig tumor cells-1 (MLTC-1) were cultured in vitro to study the functions of β2AR and M1R. The results showed that β2AR and M1R mRNA obviously expressed in mouse testis at different developing stages. β2AR and M1R mostly localized in Leydig cells of testis. NE and Ach improved the proliferation of MLTC-1 in the concentration of 10^-5 mol·L^-1 and 10^-6mol·L^-1(P<0.05). β2AR blocker butoxamine (Buto) and M1R blocker pirenzepine (Pire) both blocked the effects of NE and Ach(P<0.05). NE and Ach obviously reduced the expression of 3β-HSD mRNA in MLTC-1, and the β2AR blocker Buto and M1R blocker Pire halted the effects by blocking β2AR and M1R. NE improved the expression of ERα mRNA, but Ach enhanced the expression of ERβ mRNA, receptor blocker Buto and Pire didn't block the effects of NE and Ach. The results indicated that the functions of NE and Ach regulating the proliferation of Leydig cells and synthesis of testosterone were mediated by β2AR and M1R, but the effects of NE and Ach on the expression of ERα and ERβ were not mediated by β2AR and M1R.

中图分类号:

Q813

霍书英, 李玉荣, 武现军. 肾上腺素能受体β2和胆碱能受体M1在小鼠睾丸的表达定位及功能研究[J]. 畜牧兽医学报, 2017, 48(6): 1028-1034.

HUO Shu-ying, LI Yu-rong, WU Xian-jun. Study on the Expression and Functions of Adrenergic Receptor β2 and Muscarinic Acetylcholine Receptor M1 in Mouse Testis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(6): 1028-1034.

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参考文献

[1] YOSHIOKA K, TANAHASHI M, TAKEDA M, et al. Induction of bladder overactivity by nerve growth factor in testes in rats:possible neural crosstalk between the testes and urinary bladder[J]. Low Urin Tract Symptoms, 2016, 8(1):62-67.
[2] RAMASAMY R, STERLING J, LI P S, et al. Multiphoton imaging and laser ablation of rodent spermatic cord nerves:potential treatment for patients with chronic orchialgia[J]. J Urol, 2012, 187(2):733-738.
[3] GUTIÉRREZ-PÉREZ O, ROJAS-CASTAÑEDA J C, CHAVEZ-SALDAÑA M, et al. Infertility in rats subjected to genitofemoral nerve section is not associated with testicular damage[J]. Andrologia, 2014, 46(2):151-157.
[4] HUO S Y, XU Z H, ZHANG X X, et al. Testicular denervation in prepuberty rat inhibits seminiferous tubule development and spermatogenesis[J]. J Reprod Dev, 2010, 56(4):370-378.
[5] DIXON R A F, KOBILKA B K, STRADER D J, et al. Cloning of the gene and cDNA for mammalian β-adrenergic receptor and homology with rhodopsin[J]. Nature, 1986, 321(6065):75-79.
[6] CAULFIELD M P, BIRDSALL N J M. International union of pharmacology. XVII. Classification of muscarinic acetylcholine receptors[J]. Pharmacol Rev, 1998, 50(2):279-290.
[7] HUO S Y, ZHONG X H, WU X J, et al. Effects of norepinephrine and acetylcholine on the development of cultured Leydig cells in mice[J]. J Biomed Biotechnol, 2012, 2012:503093.
[8] CHERNOGUBOVA E, HUTCHINSON D S, NEDERGAARD J, et al. α₁- and β₁-adrenoceptor signaling fully compensates for β₃-adrenoceptor deficiency in brown adipocyte norepinephrine-stimulated glucose uptake[J]. Endocrinology, 2005, 146(5):2271-2284.
[9] ZIMRING J C, KAPP L M, YAMADA M, et al. Regulation of CD8⁺ cytolytic T lymphocyte differentiation by a cholinergic pathway[J]. J Neuroimmunol, 2005, 164(1-2):66-75.
[10] FRIAS J, RODRIGUEZ R, TORRES J M, et al. Effects of acute alcohol intoxication on pituitary-gonadal axis hormones, pituitary-adrenal axis hormones, β-endorphin and prolactin in human adolescents of both sexes[J]. Life Sci, 2000, 67(9):1081-1086.
[11] WANG G X, CHEN D W, LUO H S, et al. Low-dose ethanol suppresses 17β-estradiol activity in GH4C1 pituitary tumor cells[J]. Cell Biol Toxicol, 2010, 26(3):265-277.
[12] AKINGBEMI B T, GE R S, ROSENFELD C S, et al. Estrogen receptor-α gene deficiency enhances androgen biosynthesis in the mouse Leydig cell[J]. Endocrinology, 2003, 144(1):84-93.
[13] BAO B, KUMAR N, KARP R M, et al. Estrogen receptor-β expression in relation to the expression of luteinizing hormone receptor and cytochrome P450 enzymes in rat ovarian follicles[J]. Biol Reprod, 2000, 63(6):1747-1755.
[14] LEE W S, YOON S J, YOON T K, et al. Effects of bone morphogenetic protein-7 (BMP-7) on primordial follicular growth in the mouse ovary[J]. Mol Reprod Dev, 2004, 69(2):159-163.
[15] UEHARA T, KANEKO M, TANAKA S, et al. Possible involvement of p38 MAP kinase in HSP70 expression induced by hypoxia in rat primary astrocytes[J]. Brain Res, 1999, 823(1-2):226-230.
[16] 霍书英, 马爱进, 武现军, 等. 去精索上神经(SSN)和精索下神经(ISN)对大鼠睾丸发育和精子发生的影响[J]. 畜牧兽医学报, 2011, 42(2):260-266.
HUO S Y, MA A J, WU X J, et al. Effects of denervation of SSN and ISN on testicular development and spermatogenesis in rat[J]. Acta Veterinaria et Zootechnica Sinica, 2011, 42(2):260-266. (in Chinese).
[17] DA SILÜA JÚNIOR E D, DE SOUZA B P, RODRIGUES J Q, et al. Functional characterization of acetylcholine receptors and calcium signaling in rat testicular capsule contraction[J]. Eur J Pharmacol, 2013, 714(1-3):405-413.
[18] SCHIRMER S U, ECKHARDT I, LAU H, et al. The cholinergic system in rat testis is of non-neuronal origin[J]. Reproduction, 2011, 142(1):157-166.
[19] TROISPOUX C, REITER E, COMBARNOUS Y, et al. β2 adrenergic receptors mediate cAMP, tissue-type plasminogen activator and ransferring production in rat Sertoli cells[J]. Mol Cell Endocrinol, 1998, 142(1-2):75-86.
[20] QUEIRÓZ D B, MENDES F R, PORTO C S, et al. α₁-adrenoceptor subtypes in rat epididymis and the effects of sexual maturation[J]. Biol Reprod, 2002, 66(2):508-515.
[21] MA J K, ZHU W J. Effects of the β₂-agonist clenbuterol on testicular steroidogenic acute regulatory protein mRNA expression in adult rats[J]. J Vet Pharmacol Ther, 2010, 33(6):558-563.
[22] HARA M R, KOVACS J J, WHALEN E J, et al. A stress response pathway regulates DNA damage through β₂-adrenoreceptors and β-arrestin-1[J]. Nature, 2011, 477(7364):349-353.
[23] ADIBNIA E, RAZI M, MALEKINEJAD H. Zearalenone and 17 β-estradiol induced damages in male rats reproduction potential;evidence for ERα and ERβ receptors expression and steroidogenesis[J]. Toxicon, 2016, 120:133-146.
[24] DUMASIA K, KUMAR A, KADAM L, et al. Effect of estrogen receptor-subtype-specific ligands on fertility in adult male rats[J]. J Endocrinol, 2015, 225(3):169-180.

肾上腺素能受体β2和胆碱能受体M1在小鼠睾丸的表达定位及功能研究

Study on the Expression and Functions of Adrenergic Receptor β2 and Muscarinic Acetylcholine Receptor M1 in Mouse Testis

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