Regulation of Acetylcholine Receptors on Muscle

我们对突变斑马鱼的研究继续推进对突触传递和遗传的理解。 人类神经肌肉疾病的形式，例如肌无力综合征、慢通道综合征和布罗迪综合征， 疾病现在，我们提出的研究利用了突变株系，我们已经证明，这些突变株系要么缺乏突触， 或缺乏受体定位突触下的能力，肌肉的能力， 释放钙，肌肉螯合钙的能力，或拥有异常水平的突触驱动 表明突触前神经和突触后肌肉之间的沟通不畅。开发 通过这些线提供的优势，我们将确定突触活动在塑造两者中的作用。 突触前和突触后发育。这些研究将大大有助于利用我们最近的 开发了栓系毒素方法，以细胞自主方式沉默突触传递。 因此，我们可以确定调整突触驱动和/或单个细胞的兴奋性的后果 在运动神经元靶区内。在突变系中拯救单个突触后细胞将提供一种新的方法。 这是一种互补方法，其中使单个细胞在靶肌肉的原本沉默的区域中活跃。 这种方法的组合将被用来确定突触后活动的参与， 电耦合的发育丧失，突触前递质释放特性的获得， 建立突触后受体动力学。这项研究是可能的，凭借我们的能力，第一次， 在任何脊椎动物制备中，在体内从运动神经元和靶肌肉同时记录。这 是由于电紧凑的肌肉，允许整个细胞电压钳位和透明度， 脊髓深处运动神经元的识别和膜片钳。的组合优点 新的方法和准备现在提供了独特的机会，或审查长期存在的问题， 突触功能

Our studies of mutant zebrafish continue to advance understanding of synaptic transmission and inheritable forms of human neuromuscular disorders such as myasthenic syndrome, slow channel syndrome and Brody disease. The proposed studies now capitalize on mutant lines which we have shown either lack synaptic transmission altogether, or lack the ability of receptors to localize subsynaptically, the ability of muscle to release calcium, the ability of muscle to sequester calcium, or possess abnormal levels of synaptic drive suggesting miscommunication between presynaptic nerve and postsynaptic muscle. Exploiting the advantages provided by these lines, we will determine the role of synaptic activity in shaping both presynaptic and postsynaptic development. These studies will be greatly facilitated by use of our recently developed tethered toxin methodology that silences synaptic transmission in a cell autonomous fashion. Thus, we can determine the consequences of adjusting synaptic drive and/or excitability of individual cells within the motorneuron target field. Rescue of individual postsynaptic cells in mutant lines will provide a complementary approach wherein a single cell is rendered active in an otherwise silent field of target muscle. This combination of approaches will be used to determine the involvement of postsynaptic activity in the developmental loss of electrical coupling, acquisition of presynaptic transmitter release properties and establishment of postsynaptic receptor kinetics. The study is possible by virtue of our ability, for the first time in any vertebrate preparation, to simultaneously record from a motorneuron and target muscle in vivo. This is due to electrically compact muscle that allows whole cell voltage clamp and transparency, facilitating identification and patch clamp of motorneurons deep within the spinal cord. The combined advantages of new approaches and preparation now provide unique opportunities or examining long standing questions of synaptic function.