SYNAPSES ON TRANSGENIC MUSCLE FIBERS STUDIED OVER TIME IN LIVING MICE

描述（从摘要）：这项工作的目的是更好地 了解影响神经的目标衍生成分的作用 终端维护、消除和增长。新开发的 将外源基因和其他分子转移到活体中的技术 细胞在体内的反应和成像的修饰细胞超过 时间将用于研究神经肌肉接头。 被认为在运动终末发芽中起作用的分子的作用 将通过肌肉注射和基因 编码CNTF、BDNF、NT 3、NGF和IGF-2。同情的反应， 将探讨感觉和运动轴突以及对突触的影响。 阵蛋白质合成将在肌肉中被局部抑制， 在突触部位注射蓖麻毒素60突触前和突触后 将进行专业化。这里的争论点在于 是由于去除了最初由肌肉提供的某些因素， 突触的存活是否取决于肌肉的反应， 蛋白质合成乙酰胆碱受体（AChR）在 将通过向突触内注射 区域内有许多旨在降低突触后AChR的药物 合成.这些包括反义寡核苷酸， 反义方向和编码显性负截短的cDNA AChR和43 k蛋白的亚基。此外，还将努力 为目的基因和报告基因开发新的腺病毒载体 基因作为增加转染肌纤维产量的手段， 在原地学习。此外，亦会进行一些实地监测研究 在缺乏s-层粘连蛋白或其他基底层蛋白的转基因小鼠中， 一种研究基底层在突触消除中的作用的方法， 生长和成熟。

DESCRIPTION (from the abstract): The aim of this work is to better understand the role of target derived constituents that effect nerve terminal maintenance, elimination and growth. The newly developed techniques of transfer of exogenous genes and other molecules into living cells in vivo and the imaging of the responses of the modified cells over time will be used to study the neuromuscular junction. The role of molecules believed to play a role in motor terminal sprouting will be explored by transfecting muscles, by injection, with genes encoding CNTF, BDNF, NT3, NGF and IGF-2. The response of sympathetic, sensory and motor axons will be explored as will the effect on synapse formation. Protein synthesis will be focally inhibited in muscle by injecting ricin-60 near the synaptic site. Both pre- and postsynaptic specializations will be followed. At issue here is whether synapse loss is due to the removal of some factor initially supplied by the muscle or whether synapse survival depends on a muscle response that requires new protein synthesis. The role of the acetylcholine receptor (AChR) in synapse maintenance will be explored by injecting into the synaptic region a number of agents designed to decrease postsynaptic AChR synthesis. These include antisense oligonucleotides, cDNAs in the antisense direction and cDNAs encoding dominant negative truncations for subunits of the AChR and the 43k protein. In addition, efforts will be made to develop new adenovirus vectors for genes of interest and reporter genes as a means of increasing the yield of transfected muscle fibers to study in situ. Some in situ monitoring studies will also be conducted on transgenic mice lacking s-laminin or other basal lamina proteins as a way of investigating the role of basal lamina in synapse elimination, growth and maturation.