Nogo Receptor Signaling and Function

描述（由申请人提供）：将研究Nogo受体的信号传导机制和正常功能。最近的证据表明，Nogo受体（NogoR）通过与p75神经营养因子受体（p75 NTR）形成复合物来发出信号。存在NogoR和p75 NTR两者的未表征的结构同源物。将确定这些同源物是否也形成复合物，以及它们是否通过RhoA和NF-κ B的Nogo或MAG依赖性激活与NogoR/p75 NTR复合物类似地发出信号。这些研究将涉及小鼠和斑马鱼NogoR同源物的分子克隆，以及转染的NogoR、NogoR同源物、p75 NTR和p75 NTR同源物在培养的哺乳动物细胞系中的表达。先前的研究表明NogoR/p75 NTR信号传导功能抑制损伤的成年脊髓的再生。据推测，NogoR/p75 NTR和同源物在胚胎发育过程中也起作用，以引导神经轴突到达其适当的靶标。这一假设将通过采用吗啉代反义RNA技术来破坏这些蛋白在斑马鱼胚胎中的表达来进行测试。这些操作对轴突靶向的影响将通过将荧光标记物Dil注射到胚胎中以允许轴突轨迹的显微成像来评估。

DESCRIPTION (provided by applicant): The signaling mechanisms and normal functions of the Nogo receptor will be investigated. Recent evidence indicates that the Nogo receptor (NogoR) signals by forming a complex with the p75 neurotrophin receptor (p75NTR). Uncharacterized structural homologs of both NogoR and p75NTR exist. It will be determined whether these homologs also form complexes and whether they signal similarly to the NogoR/p75NTR complex, via Nogo or MAG-dependent activation of RhoA and NF-kappaB. These studies will involve molecular cloning of mouse and zebrafish NogoR homologs, and expression in cultured mammalian cell lines of transfected NogoR, NogoR homolog, p75NTR and p75NTR homologs. Previous studies indicate that NogoR/p75NTR signaling functions to inhibit regeneration of the injured adult spinal cord. It is postulated that NogoR/p75NTR and homologs also function during embryonic development to guide neural axons to their appropriate targets. This hypothesis will be tested by employing morpholino-antisense RNA technology to disrupt expression of these proteins in zebrafish embryos. Effects of these manipulations on axonal targeting will be assessed by injecting the fluorescent marker Dil into embryos to allow microscopic imaging of axon trajectories.