Eich Marie-Lisa, Dembla Ekta, Wahl Silke, Dembla Mayur, Schwarz Karin, Schmitz Frank
Department of Neuroanatomy, Medical School Homburg/Saar, Institute for Anatomy and Cell Biology, Saarland UniversityHomburg/Saar, Germany.
Front Mol Neurosci. 2017 Jul 25;10:230. doi: 10.3389/fnmol.2017.00230. eCollection 2017.
In the present study, we generated and characterized a splice site-specific monoclonal antibody that selectively detects the calcineurin-binding dynamin1 splice variant dynamin1xb. Calcineurin is a Ca-regulated phosphatase that enhances dynamin1 activity and is an important Ca-sensing mediator of homeostatic synaptic plasticity in neurons. Using this dynamin1xb-specific antibody, we found dynamin1xb highly enriched in synapses of all analyzed brain regions. In photoreceptor ribbon synapses, dynamin1xb was enriched in close vicinity to the synaptic ribbon in a manner indicative of a peri-active zone immunolabeling. Interestingly, in dark-adapted mice we observed an enhanced and selective enrichment of dynamin1xb in both synaptic layers of the retina in comparison to light-adapted mice. This could be due to an illumination-dependent recruitment of dynamin1xb to retinal synapses and/or due to a darkness-induced increase of dynamin1xb biosynthesis. These latter findings indicate that dynamin1xb is part of a versatile and highly adjustable, activity-regulated endocytic synaptic machinery.
在本研究中,我们制备并鉴定了一种剪接位点特异性单克隆抗体,该抗体可选择性检测与钙调神经磷酸酶结合的发动蛋白1剪接变体发动蛋白1xb。钙调神经磷酸酶是一种钙调节磷酸酶,可增强发动蛋白1的活性,是神经元中稳态突触可塑性的重要钙传感介质。使用这种针对发动蛋白1xb的特异性抗体,我们发现发动蛋白1xb在所有分析脑区的突触中高度富集。在光感受器带状突触中,发动蛋白1xb以一种表明活性区周围免疫标记的方式富集在突触带附近。有趣的是,在暗适应小鼠中,与光适应小鼠相比,我们观察到发动蛋白1xb在视网膜的两个突触层中均有增强的选择性富集。这可能是由于发动蛋白1xb受光照依赖募集到视网膜突触,和/或由于黑暗诱导发动蛋白1xb生物合成增加。这些最新发现表明,发动蛋白1xb是一种多功能、高度可调节的、活性调节的内吞突触机制的一部分。