Hypothesis Relating the Structure, Biochemistry and Function of Active Zone Material Macromolecules at a Neuromuscular Junction

This report integrates knowledge of in situ macromolecular structures and synaptic protein biochemistry to propose a unified hypothesis for the regulation of certain vesicle trafficking events (i.e., docking, priming, Ca2+-triggering, and membrane fusion) that lead to neurotransmitter secretion from specialized “active zones” of presynaptic axon terminals. Advancements in electron tomography, to image tissue sections in 3D at nanometer scale resolution, have led to structural characterizations of a network of different classes of macromolecules at the active zone, called “Active Zone Material’. At frog neuromuscular junctions, the classes of Active Zone Material macromolecules “top-masts”, “booms”, “spars”, “ribs” and “pins” direct synaptic vesicle docking while “pins”, “ribs” and “pegs” regulate priming to influence Ca2+-triggering and membrane fusion. Other classes, “beams”, “steps”, “masts”, and “synaptic vesicle luminal filaments’ likely help organize and maintain the structural integrity of active zones. Extensive studies on the biochemistry that regulates secretion have led to comprehensive characterizations of the many conserved proteins universally involved in these trafficking events. Here, a hypothesis including a partial proteomic atlas of Active Zone Material is presented which considers the common roles, binding partners, physical features/structure, and relative positioning in the axon terminal of both the proteins and classes of macromolecules involved in the vesicle trafficking events. The hypothesis designates voltage-gated Ca2+ channels and Ca2+-gated K+ channels to ribs and pegs that are connected to macromolecules that span the presynaptic membrane at the active zone. SNARE proteins (Syntaxin, SNAP25, and Synaptobrevin), SNARE-interacting proteins Synaptotagmin, Munc13, Munc18, Complexin, and NSF are designated to ribs and/or pins. Rab3A and Rabphillin-3A are designated to top-masts and/or booms and/or spars. RIM, Bassoon, and Piccolo are designated to beams, steps, masts, ribs, spars, booms, and top-masts. Spectrin is designated to beams. Lastly, the luminal portions of SV2 are thought to form the bulk of the observed synaptic vesicle luminal filaments. The goal here is to help direct future studies that aim to bridge Active Zone Material structure, biochemistry, and function to ultimately determine how it regulates the trafficking events in vivo that lead to neurotransmitter secretion.

(M)Unc13s in Active Zone Diversity: A Drosophila Perspective

The so-called active zones at pre-synaptic terminals are the ultimate filtering devices, which couple between action potential frequency and shape, and the information transferred to the post-synaptic neurons, finally tuning behaviors. Within active zones, the release of the synaptic vesicle operates from specialized “release sites.” The (M)Unc13 class of proteins is meant to define release sites topologically and biochemically, and diversity between Unc13-type release factor isoforms is suspected to steer diversity at active zones. The two major Unc13-type isoforms, namely, Unc13A and Unc13B, have recently been described from the molecular to the behavioral level, exploiting Drosophila being uniquely suited to causally link between these levels. The exact nanoscale distribution of voltage-gated Ca2+ channels relative to release sites (“coupling”) at pre-synaptic active zones fundamentally steers the release of the synaptic vesicle. Unc13A and B were found to be either tightly or loosely coupled across Drosophila synapses. In this review, we reported recent findings on diverse aspects of Drosophila Unc13A and B, importantly, their nano-topological distribution at active zones and their roles in release site generation, active zone assembly, and pre-synaptic homeostatic plasticity. We compared their stoichiometric composition at different synapse types, reviewing the correlation between nanoscale distribution of these two isoforms and release physiology and, finally, discuss how isoform-specific release components might drive the functional heterogeneity of synapses and encode discrete behavior.

SEISMICITY of CENTRAL ASIA in 2015 (Kyrgyzstan, Uzbekistan and Southeastern Kazakhstan)

Seismic observations on the territory of Central Asia in 2015 were carried out by monitoring systems of three countries: Kyrgyzstan, Uzbekistan and Kazakhstan consisting of 25, 29 stations of the first two countries and 43 stations and five seismic groups of the third country respectively. The catalogue of earthquakes in 2015 included 302 events with KR=8.614.1. The largest earthquake in the catalogue with KR=14.1 was registered on November 17, 2015, at 17h 29m in the Osh area of Kyrgyzstan, near the Taldyk settlement. It was accompanied by numerous aftershocks (852) with KR=4.111.4. In general, the seismic situation in the region in 2015 is characterized by lower values of the number of earthquakes N and the released seismic energy E compared to the average annual values of these parameters for the period from 1996 to 2014. The location and configuration of the main seismically active zones remained without changes.

Reconstructing essential active zone functions within a synapse

Active zones are molecular machines that control neurotransmitter release through synaptic vesicle docking and priming, and through coupling of these vesicles to Ca2+ entry. The complexity of active zone machinery has made it challenging to determine which mechanisms drive these roles in release. Here, we induce RIM+ELKS knockout to eliminate active zone scaffolding networks, and then reconstruct each active zone function. Re-expression of RIM1-Zn fingers positioned Munc13 on undocked vesicles and rendered them release-competent. Reconstitution of release-triggering required docking of these vesicles to Ca2+ channels. Fusing RIM1-Zn to CaVbeta4-subunits sufficed to restore docking, priming and release-triggering without reinstating active zone scaffolds. Hence, exocytotic activities of the 80 kDa CaVbeta4-Zn fusion protein bypassed the need for megadalton-sized secretory machines. These data define key mechanisms of active zone function, establish that fusion competence and docking are mechanistically separable, and reveal that active zone scaffolding networks are not required for release.

Enhancement of wavefront measurement sensitivity in a zonal wavefront sensor without curtailing the sensing speed

In this paper we propose a zonal wavefront sensing scheme that facilitates wavefront measurement with enhanced sensitivity at the standard video rate. We achieve this enhanced sensitivity by implementing a sequential display of binary holograms described over each zone sampling the incident wavefront with the help of a ferroelectric liquid crystal spatial light modulator. By keeping the number of active zones as 24 and using a camera with an imaging frame rate equal to the binary hologram display rate of the spatial light modulator, we are able to reach the sensing frame rate of 60 Hz. In addition to enhancement in sensitivity, the proposed scheme facilitates zone wise tuning of binary holograms and eliminates the possibility of any crosstalk between adjacent zones. We perform a proof-of-principle experiment that validates the proposed zonal wavefront sensing scheme and demonstrates its advantages.

Methods and results of long-term strong earthquakes forecast in the Uzbekistan territory

An approach to evaluate the current seismological situation in the Uzbekistan territory is presented. This approach is based on the regularities of seismic processes in strong-earthquake focal areas and the manifestation peculiarities of strong earthquakes in seismically active zones. At the first stage, within seismically active zones, areas with a high seismic activity matching the strong earthquake level were identified during the historical and instrumental observation periods. Considering the low variability in the direction of seismotectonic processes over tens and hundreds of years, which determines the modern stress state of seismically active structures, these areas were considered the most likely areas to experience strong earthquakes over the next few decades. Tectonophysical validation of the division of seismically active zones into areas with different potential hazards of strong earthquakes was carried out within the framework of cataclastic analysis method of rupture dislocations (CAM). At the second stage, temporal fluctuations in seismic regime parameters within the selected areas were studied. Based on the number of current anomalous features identified, the areas were ranked according to the occurrence probability of strong earthquakes over the next few years.

Hydrothermal Ore Genesis in the Sea of Japan

—The Sea of Japan is a tectonically active region with rift-related destruction of the Earth’s crust and numerous volcanic edifices on the seafloor. Since the 1970s, numerous zones with ferromanganese crusts (FMCs) and phosphorite and barite ore occurrences have been discovered during the repeated expeditions of the Pacific Oceanological Institute, Vladivostok. Analysis of the distribution of these ore occurrences showed that all of them are confined to tectonically active zones of the seafloor: submarine volcanoes, tectonic scarps, or fault zones. In some zones, phosphorites occur together with FMCs, and in one zone, together with FMCs and barites. Ferromanganese hydroxides, phosphorites, or barites are found in the pores of basalts composing submarine volcanic edifices in the Sea of Japan. These data indicate that the ore matter in all zones is supplied with postvolcanic gas-hydrothermal fluids or hydrothermal solutions circulating along deep faults during the destruction of the continental crust in the southern and eastern parts of the sea. Thus, ferromanganese, phosphate, and barite ore occurrences in the Sea of Japan are related to low-temperature hydrothermal-sedimentary processes.

Cooperative Amyloid Fibre Binding and Disassembly by the Hsp70 disaggregase

Although amyloid fibres are highly stable protein aggregates, a specific combination of human Hsp70 system chaperones can disassemble them, including fibres formed of α-synuclein, huntingtin or Tau. Disaggregation requires the ATPase activity of the constitutively expressed Hsp70, Hsc70, together with the J domain protein DNAJB1 and the nucleotide exchange factor Apg2. Recruitment and clustering of Hsc70 on the fibrils appear to be necessary for disassembly. Here we use atomic force microscopy (AFM) to show that segments of in vitro assembled α-synuclein fibrils are first coated with chaperones and then undergo bursts of rapid, unidirectional disassembly. Cryo-electron tomography reveals fibrils with regions of densely bound chaperones extending from the fibre surface, preferentially at one end of the fibre. Sub-stoichiometric amounts of Apg2 relative to Hsc70 dramatically increase recruitment of Hsc70 to the fibres, creating localised active zones that then undergo rapid disassembly at a rate of ~4 subunits per second.