Fetal Testicular Maturation by Estrogen--Impact on Adult Fertility

雌激素促进胎儿睾丸成熟--对成年生育能力的影响

• 批准号: 7633362
• 负责人: Eugene D. Albrecht
• 金额: $ 38.99万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7633362
• 关键词: Adult; Apoptosis; Aromatase; Aromatase Inhibitors; Cell Proliferation; Cells; Collaborations; Competence; Coupled; Development; Duct (organ) structure; Enzymes; Estradiol; Estrogen Receptor alpha; Estrogen Receptors; Estrogens; Exhibits; Fertility; Fetal Development; Fetus; Figs - dietary; Follicle Stimulating Hormone Receptor; Foundations; Germ; Human; Human Development; Infertility; Knockout Mice; Knowledge; Life; Male Genital Organs; Messenger RNA; Modeling; Mothers; Neonatal; Ovarian; Papio; Pregnancy; Primates; Process; Proteins; Regulation; Reproductive Endocrinology; Reverse Transcriptase Polymerase Chain Reaction; Structure of efferent ductule of testis; Testing; Testis; Translating; Universities; Virginia; day; fetal; human male; immunocytochemistry; in utero; in vivo; laser capture microdissection; male; medical schools; neonate; nonhuman primate; programs; reproductive; reproductive axis; reproductive function; research study

Description (provided by applicant): The foundation for development of the human and nonhuman primate male reproductive system occurs in utero and thus improper development of the testes during fetal life may compromise reproductive function and fertility in adulthood. Very little is known, however, about the regulation of human male fetal reproductive maturation because for ethical reasons it is difficult to study this process in humans. We have used the baboon as a nonhuman primate model and shown that estrogen regulates fetal ovarian development (Project I). Estrogen receptor (ER) or P-450 aromatase null mice exhibit abnormal testicular development and infertility. Therefore, we propose to use the baboon to test the hypotheses that: (1) estrogen regulates maturation of the primate fetal testes and/or efferent ducts in utero and (2) this estrogen-dependent programming of the male fetus determines reproductive function and fertility in adulthood. In Experiment 1, normal development of the testes and efferent ducts will be determined in fetal baboons obtained on days 60 (early), 100 (mid) and 165 (late) of gestation (term = 184 days) in association with the increase in estrogen of pregnancy. In Experiment 2, testes development will be assessed on day 165 of gestation in fetal baboons in which estrogen levels are suppressed by maternal administration of the aromatase inhibitor CGS 20267 throughout the second half of pregnancy. Development will be assessed by morphometric analysis of germ, Sertoli, and efferent ductule/epididymal cells, and cell proliferation/apoptosis. Function will be assessed by RT-PCR quantification of mRNA levels for ER alpha and beta , steroidogenic enzymes (e.g., P-450 aromatase) and other regulatory molecules (e.g., FSH receptor) in testicular cells isolated by laser capture microdissection, and determining cellular expression of respective proteins in our Immunocytochemistry Core. In Experiment 3, neonates delivered from baboon mothers treated with CGS 20267 estradiol throughout the second half of gestation will be reared to adulthood to determine impact on reproductive competence of the adult male. This project will be carried out in collaboration with experts in primate fetal development and male reproductive endocrinology at the Eastern Virginia Medical School, University of Pittsburgh SCCPRR, and Johns Hopkins University SCCPRR. The collaborative in vivo experimental approach in the primate, coupled with the ability to rear estrogen-deprived neonatal baboons to adulthood, provide a unique opportunity to study programming of the male reproductive axis. Results are expected to translate to the human and thus advance knowledge on the regulation of reproductive maturation and fertility in the human male.

描述（由申请人提供）：人类和非人类灵长类动物男性生殖系统的发展基础发生在子宫内，因此在胎儿寿命期间的睾丸发育不当可能会损害成年后的生殖功能和生育能力。然而，关于人类男性胎儿生殖成熟的调节鲜为人知，因为出于道德原因，很难在人类中研究这一过程。我们已经将狒狒用作非人类灵长类动物模型，并表明雌激素调节胎儿卵巢发育（项目I）。雌激素受体（ER）或P-450芳香酶无小鼠表现出异常的睾丸发育和不育。因此，我们建议使用狒狒测试：（1）雌激素调节子宫内雌激素胎儿睾丸的成熟和/或传染性导管的成熟，以及（2）雄性胎儿的这种雌激素依赖性编程确定成年期生殖功能和生育能力。在实验1中，正常发展 睾丸和传出导管将在第60天（早期），100（中）和妊娠165（后期）获得的胎儿狒狒中确定（期限= 184天），以及妊娠雌激素的增加。在实验2中，将在胎儿狒狒的妊娠第165天评估睾丸发育，在该胎儿狒狒中，雌激素水平在整个妊娠的下半年都通过孕妇抑制剂CGS 20267抑制。发育将通过对细菌，Sertoli和Fuderent ductule/Epididymal细胞以及细胞增殖/凋亡的形态分析进行评估。功能将通过RT-PCR定量对ERα和β，类固醇生成酶（例如P-450芳香酶）和其他调节分子（例如，FSH受体）在激光捕获微异常中分离出的睾丸细胞中的睾丸细胞中的其他调节分子（例如FSH受体）评估功能。在实验3中，从妊娠的后半部分接受CGS 20267雌二醇治疗的狒狒母亲分娩的新生儿将饲养到成年后，以确定对成年男性生殖能力的影响。该项目将在合作中进行 在弗吉尼亚州东部医学院，匹兹堡大学SCCPRR和约翰·霍普金斯大学SCCPRR上，具有灵长类动物胎儿发育专家和男性生殖内分泌学专家。灵长类动物中的协作体内实验方法，再加上雌激素剥夺的新生儿狒狒的成年能力，为研究男性生殖轴的编程提供了独特的机会。预期结果将转化为人类，因此可以提高人们对人类男性生殖成熟和生育能力的了解。