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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 性发育中的事件顺序是非常重要的。不适当的荷尔蒙环境会导致成年男性不育。大约15%的夫妇受到不孕症的影响，其中40%是男性不育。了解男性性发育的荷尔蒙调节可能有助于及早发现和预防不孕症。该计划的目标是了解成熟过程中垂体促性腺激素差异表达的调节。有资料表明，垂体腺苷环化酶激活多肽(PACAP)对垂体促性腺激素具有差异性调节作用。PACAP增加促性腺激素释放激素(GnRH)的作用，单独PACAP促进黄体生成素的产生，同时降低卵泡刺激素的表达。我们的总体假设是，在胚胎和出生后早期发育过程中，脑垂体暴露在非常高水平的PACAP中，选择性地抑制FSH，直到性成熟的幼年阶段。这项建议的具体目的是：1)确定PACAP是否在胎儿垂体发育过程中促性腺激素亚单位基因的差异表达中发挥作用；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)确定胎儿和幼年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；以及4)通过PACAP拮抗剂PACAP-(6-38)的垂体特异性表达来探讨PACAP对发育的影响。拟议中的实验将在垂体细胞培养、原位杂交、激光捕获显微切割、单细胞RT-PCR、原位杂交、Northern杂交、Southern杂交和cDNA克隆等技术方面发展皮？S的技能。这项建议的成功完成将提供对促性腺激素发育调节的洞察和宝贵的经验，应聘者可以利用这些经验来宣传成功的职业生涯。

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.