COBRE: UL: P4: DVMT PROCESSES OF PITUITARY DIFFERENTIATION & MATURATION

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。性发育的顺序是非常重要的。不适当的荷尔蒙环境会导致成年男性不育。不育影响了大约15%的夫妇，其中40%是男性不育。了解男性性发育的激素调节可能有助于早期发现和预防不育症。该计划的目标是发展对成熟过程中垂体促性腺激素差异表达调控的理解。有研究表明垂体腺苷酸环化酶激活多肽（PACAP）对垂体促性腺激素有差异调节作用。PACAP增加促性腺激素释放激素（GnRH）的作用，单独使用PACAP可增加促黄体生成素的产生，同时降低促卵泡激素的表达。我们的总体假设是，在胚胎和出生后早期发育期间，垂体暴露于非常高水平的PACAP，选择性地抑制FSH，直到性成熟的少年阶段。本研究的具体目的是：1)确定在胎儿垂体发育过程中，PACAP是否在促性腺激素亚基基因的差异表达中起作用；2)确定未成熟雄性大鼠FSHb mRNA的升高是否与断奶有关，并与PACAP表达的变化一致；3)确定胎鼠和幼龄雄性大鼠的垂体细胞是否比幼年大鼠的垂体细胞对PACAP更敏感；4)通过垂体特异性表达PACAP拮抗剂PACAP-，探讨PACAP对发育的影响（6-38）。本实验将发展PI在垂体细胞培养、原位杂交、激光捕获显微解剖、单细胞RT-PCR、原位杂交、Northern杂交、Southern杂交和cDNA克隆等技术方面的技能。成功完成该提案将为促性腺激素的发育调节提供见解，并为候选人提供宝贵的经验，以促进成功的职业生涯。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The sequence of events in sexual development is very important. An inappropriate hormonal environment can result in infertility of the adult male. Infertility affects approximately 15% of couples, 40% are male infertility. An understanding of hormonal regulation of male sexual development may allow for early detection and prevention of infertility. The goals of this plan are to develop an understanding of regulation of the differential expression of the pituitary gonadotropins during maturation. There is data suggesting that pituitary adenylate cyclase activating polypeptide (PACAP) can differentially regulate the pituitary gonadotropins. PACAP increases the actions of gonadotropin releasing hormone (GnRH) and alone PACAP enhances the production of luteinizing hormone while decreasing follicle stimulating hormone expression. Our overall hypothesis is that during embryonic and early postnatal development, the pituitary is exposed to very high levels of PACAP that selectively suppress FSH until the juvenile stage of sexual maturity. The specific aims of this proposal are: 1) To determine whether PACAP plays a role in the differential expression of the gonadotropin subunit genes during fetal pituitary development; 2) To determine if the rise in FSHb mRNA in immature male rats is related to weaning, and is coincident with a change in PACAP expression; 3) To determine if pituitary cells from fetal and infant male rats are more responsive to PACAP than are cells from juvenile rats; and 4) To explore the developmental effects of PACAP through pituitary specific expression of the PACAP antagonist PACAP-(6-38). The proposed experiments will develop the PI¿s skills in the techniques of pituitary cell culture, in situ hybridization, laser capture microdissection, single cell RT-PCR, in situ hybridization, Northern hybridization, Southern hybridization, and cDNA cloning. Successful completion of this proposal should provide insight into developmental regulation of gonadotropins and invaluable experience that the candidate can utilize to propagate a successful career.