Effects of fetal and neonatal exposure of male mice to diethylstilbestrol as endocrine disrupter on the reproductive and neuroendocrine system

雄性小鼠胎儿和新生儿暴露于己烯雌酚作为内分泌干扰物对生殖和神经内分泌系统的影响

• 批准号: 12836014
• 负责人: HOSHI Nobuhiko
• 金额: $ 2.05万
• 依托单位: Kitasato University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12836014/
• 关键词: Endcrine disrupter chemicals; Sex determination; Sex differentiation; diethylstilbestrol; spermatogenesis; mouse; hypothalamus- hypophysis-gonadal axis; 内分泌撹乱化学物質; 脳-下垂体-性腺系

The purpose of this study is to determine the effect of fetal and neonatal exposure to diethylstilbestrol (DBS), as a representative endocrine disrupter, on the reproductive function of male mice. Various doses (0.001-100 μg) of DES or a vehicle (control) were administered to the neonatal group on day 3 (single) and on every 2 days from day 3 (N-long-term), and to the fetal group every 2 days from 7 dpc (F-long-term). Endocrinological and morphological changes were evaluated at 25 days of age (immediately before the onset of puberty), 4, 6, 8, and 12 weeks of age. We established a non-RI solid phase immunoassay method, which enables multiple microassay (LH, FSH, PRL, testosterone, inhibin) of the same sample with high sensitivity and specificity. In single administration of 10 and 100 μg of DES at 4 weeks, N-long-term administration at 1.0 μg for 25 days and 10 μg for 6 weeks, aid F-long-term administration at 10 μg for 6 weeks, testis weights were significantly reduced, and no spermat … More ogenesis and immature seminiferous tubules were observed. DES injections caused dose-dependent retardation of pituitary LH, FSH. Using the morphometric immunodetection method for incorporated thymidine analog BrdU, the high-dose groups (10 ＆ 100 μg) showed a marked disruption of sperm production and significant suppression of cellular proliferation in testes. Immunohistochemical analysis in each group showed that immunoreactive neurons for GnRH were generally fewer in number and had thinner neurites in the high-dose groups Immunoreactivity of Era and StAR (steroidogenic acute regulatory protein) in testes was decreased. Thus our findings indicate that 1) single neonatal exposure of male mice to 10 μg of DES can advance the spermatogenesis at 6 weeks of age; 2) long-term exposure of these mice, particularly from fetus, could not recover the spermatogenesis even under 1.0 μg of DES; and 3) there is threshold dose of DES, and these delay and damage in the reproduction system due to reduction of pituitary gonadotropin production. Less

本研究的目的是确定胎儿和新生儿接触己烯雌酚（DBS）（作为一种代表性内分泌干扰物）对雄性小鼠生殖功能的影响。不同剂量（0.001-100 μg）的 DES 或载体（对照）。 ）在第 3 天（单次）和从第 3 天起每 2 天（N-长期）给予新生儿组，从第 7 天起每 2 天给予胎儿组dpc（F-长期）在 25 天（青春期即将开始之前）、4、6、8 和 12 周龄时评估内分泌和形态学变化。方法，可在单次给药 10 和 100 次时对同一样品进行多种微量测定（LH、FSH、PRL、睾酮、抑制素）。 μg DES 4周，N-长期给药1.0 μg 25天和10 μg 6周，Aid F-长期给药10 μg 6周，睾丸重量显着减轻，无精子……使用形态学免疫检测方法观察到更多的生精小管注射引起垂体 LH、FSH 的剂量依赖性延迟。对于掺入的胸苷类似物 BrdU，高剂量组（10 和 100 μg）显示出精子生成的显着破坏以及睾丸中细胞增殖的显着抑制。免疫组织化学分析表明，每组中 GnRH 的免疫反应神经元数量普遍较少，并且睾丸细胞增殖受到显着抑制。高剂量组的神经突更薄，测试中 Era 和 StAR（类固醇生成急性调节蛋白）的免疫反应性降低，因此我们的研究结果表明 1)。雄性新生小鼠单次暴露于 10 μg DES 可以促进 6 周龄时的精子发生；2) 这些小鼠，特别是胎儿的长期暴露，即使在 1.0 μg DES 下也无法恢复精子发生；3) DES 存在阈剂量，并且由于垂体促性腺激素产生减少而导致生殖系统的延迟和损害。

项目成果

Yamamoto R: "Lens culinaris agglutinin-reactive α-fetoprotein, an alternative variant to α-fetoprotein in prenatal screening for Down's syndrome"Hum Reprod. 16・11. 2438-2444 (2001)

Yamamoto R：“Lens culinaris 凝集素反应性 α-胎蛋白，唐氏综合症产前筛查中的 α-胎蛋白的替代变体”Hum Reprod 16・11 (2001)。

Sugawara T et al.: "Sp1 and SF-1 interact and cooperate in the regulation of human steroidogenic acute regulatory protein gene expression"Endocrinology. 141. 2895-2903 (2000)

Sukawara T 等人：“Sp1 和 SF-1 在人类类固醇生成急性调节蛋白基因表达的调节中相互作用和配合”内分泌学。

Sugawara T et al.: "RIP 140 modulates transcription of the steroidogenic acute regulatory protein gene through interactions with both SF-1 and DAX-1"Endocrinology. 142. 3570-3577 (2001)

Sukawara T 等人：“RIP 140 通过与 SF-1 和 DAX-1 的相互作用调节类固醇生成急性调节蛋白基因的转录”内分泌学。

Sugawara T: "The effect of the aryihydrocarbon receptor on the human steroidogenie acute regulatory gene promoter activity"Steroid Biochem Mol Biol. 78. 253-260 (2001)

Sukawara T：“芳基烃受体对人类类固醇生成急性调节基因启动子活性的影响”Steroid Biochem Mol Biol。

Yamamoto R et al.: "Lens culinaris agglutinin-reactive α-fetoprotein, an alternative variant to a-fetoprotein in prenatal screening for Down's syndrome"Hum Reprod. 16. 2438-2444 (2001)

Yamamoto R 等人：“Lens culinaris 凝集素反应性 α-胎蛋白，唐氏综合症产前筛查中甲胎蛋白的替代变体”Hum Reprod. 16. 2438-2444 (2001)