Drosophila Spastin Regulates Synaptic Microtubule Networks and Is Required for Normal Motor Function

Nina Tang Sherwood is with California Institute of Technology, Qi Sun is with California Institute of Technology, Mingshan Xue is with UT Austin, Bing Zhang is with UT Austin, Kai Zinn is with California Institute of Technology.The most common form of human autosomal dominant hereditary spastic paraplegia (AD-HSP) is caused by mutations in the SPG4 (spastin) gene, which encodes an AAA ATPase closely related in sequence to the microtubule-severing protein Katanin. Patients with AD-HSP exhibit degeneration of the distal regions of the longest axons in the spinal cord. Loss-of-function mutations in the Drosophila spastin gene produce larval neuromuscular junction (NMJ) phenotypes. NMJ synaptic boutons in spastin mutants are more numerous and more clustered than in wild-type, and transmitter release is impaired. spastin-null adult flies have severe movement defects. They do not fly or jump, they climb poorly, and they have short lifespans. spastin hypomorphs have weaker behavioral phenotypes. Overexpression of Spastin erases the muscle microtubule network. This gain-of-function phenotype is consistent with the hypothesis that Spastin has microtubule-severing activity, and implies that spastin loss-of-function mutants should have an increased number of microtubules. Surprisingly, however, we observed the opposite phenotype: in spastin-null mutants, there are fewer microtubule bundles within the NMJ, especially in its distal boutons. The Drosophila NMJ is a glutamatergic synapse that resembles excitatory synapses in the mammalian spinal cord, so the reduction of organized presynaptic microtubules that we observe in spastin mutants may be relevant to an understanding of human Spastin's role in maintenance of axon terminals in the spinal cord.Biological Sciences, School o

Dueling Ca2+ Sensors in Neurotransmitter Release

Ca2+-triggered neurotransmitter release is characterized by two kinetically distinct components: a fast synchronous phase and a slow asynchronous phase. Yao et al. (2011) now report that double C2 domain (Doc2) proteins function as high-affinity Ca2+ sensors to specifically regulate the asynchronous component of neurotransmitter release

Structurally and functionally unique complexins at retinal ribbon synapses

Ribbon synapses in retinal sensory neurons maintain large pools of readily releasable synaptic vesicles. This allows them to release several hundreds of vesicles per second at every presynaptic release site. The molecular components that cause this high transmitter release efficiency of ribbon synapses are unknown. In the present study, we identified and characterized two novel vertebrate complexins (CPXs), CPXs III and IV, that are the only CPX isoforms present in retinal ribbon synapses. CPXs III and IV are COOH-terminally farnesylated, and, like CPXs I and II, bind to SNAP receptor complexes. CPXs III and IV can functionally replace CPXs I and II, and their COOH-terminal farnesylation regulates their synaptic targeting and modulatory function in transmitter release. The novel CPXs III and IV may contribute to the unique release efficacy of retinal sensory neurons

Structural and Mutational Analysis of Functional Differentiation between Synaptotagmins-1 and -7

Synaptotagmins are known to mediate diverse forms of Ca2+-triggered exocytosis through their C2 domains, but the principles underlying functional differentiation among them are unclear. Synaptotagmin-1 functions as a Ca2+ sensor in neurotransmitter release at central nervous system synapses, but synaptotagmin-7 does not, and yet both isoforms act as Ca2+ sensors in chromaffin cells. To shed light into this apparent paradox, we have performed rescue experiments in neurons from synaptotagmin-1 knockout mice using a chimera that contains the synaptotagmin-1 sequence with its C2B domain replaced by the synaptotagmin-7 C2B domain (Syt1/7). Rescue was not achieved either with the WT Syt1/7 chimera or with nine mutants where residues that are distinct in synaptotagmin-7 were restored to those present in synaptotagmin-1. To investigate whether these results arise because of unique conformational features of the synaptotagmin-7 C2B domain, we determined its crystal structure at 1.44 Å resolution. The synaptotagmin-7 C2B domain structure is very similar to that of the synaptotagmin-1 C2B domain and contains three Ca2+-binding sites. Two of the Ca2+-binding sites of the synaptotagmin-7 C2B domain are also present in the synaptotagmin-1 C2B domain and have analogous ligands to those determined for the latter by NMR spectroscopy, suggesting that a discrepancy observed in a crystal structure of the synaptotagmin-1 C2B domain arose from crystal contacts. Overall, our results suggest that functional differentiation in synaptotagmins arises in part from subtle sequence changes that yield dramatic functional differences

Mechanically Robust and Repairable Superhydrophobic Zinc Coating via a Fast and Facile Method for Corrosion Resisting

Zinc coatings and superhydrophobic surfaces have their own characteristics in terms of metal corrosion resistance. Herein, we have prepared a robust and repairable superhydrophobic zinc coating (SZC) based on a widely commercially available cold galvanized paint via a fast (within 10 min) and facile process for corrosion resistance. Specifically, the cold galvanized paint was sprayed onto the iron substrate, followed by acetic acid (HAc) etching and stearic acid (STA) hydrophobizing. The as-obtained sample was coded as Fe-Zn-HAc-STA and possessed an apparent contact angle of 168.4 ± 1.5° as well as a sliding angle of 3.5 ± 1.2°. The Fe-Zn-HAc-STA sample was mechanically durable and easily repairable. After being ultrasonicated in ethanol for 100 min, the superhydrophobicity was still retained. The Fe-Zn-HAc-STA sample lost its superhydrophobicity after being abraded against sandpaper with a load of 100 g and regained its superhydrophobicity after HAc etching and subsequent STA hydrophobizing. The corrosion resistance of the SZC was investigated by immersing the Fe-Zn-HAc-STA sample into the static or dynamic aqueous solution of NaCl (3.5 wt.%) and the lasting life of the entrapped underwater air layer (EUAL) was roughly determined by the turning point at the variation curve of surface wettability against immersion time. The lasting life of the EUAL iwas 8 to 10 days for the SZC in the static NaCl solution and it decreased sharply to 12 h in a dynamic one with the flow rate of 2 and 4 m/s. This suggests that the superhydrophobic surface provided extra corrosion protection of 8 to 10 days or 12 h to the zinc coating. We hope that the SZC may find its practical application due to the facile and fast fabrication procedure, the good mechanical durability, the easy repairability, and the good corrosion protection