Drosophila TRPA Channel Painless Inhibits Male–Male Courtship Behavior through Modulating Olfactory Sensation

The Drosophila melanogaster TRPA family member painless, expressed in a subset of multidendritic neurons embeding in the larval epidermis, is necessary for larval nociception of noxious heat or mechanical stimuli. However, the function of painless in adult flies remains largely unknown. Here we report that mutation of painless leads to a defect in male–male courtship behavior and alteration in olfaction sensitivity in adult flies. Specific downregulation of the expression of the Painless protein in the olfactory projection neurons (PNs) of the antennal lobes (ALs) resulted in a phenotype resembling that found in painless mutant flies, whereas overexpression of Painless in PNs of painless mutant males suppressed male–male courtship behavior. The downregulation of Painless exclusively during adulthood also resulted in male–male courtship behavior. In addition, mutation of the painless gene in flies caused changes in olfaction, suggesting a role for this gene in olfactory processing. These results indicate that functions of painless in the adult central nervous system of Drosophila include modulation of olfactory processing and inhibition of male–male courtship behavior

Precise Spatiotemporal Control of Optogenetic Activation Using an Acousto-Optic Device

Light activation and inactivation of neurons by optogenetic techniques has emerged as an important tool for studying neural circuit function. To achieve a high resolution, new methods are being developed to selectively manipulate the activity of individual neurons. Here, we report that the combination of an acousto-optic device (AOD) and single-photon laser was used to achieve rapid and precise spatiotemporal control of light stimulation at multiple points in a neural circuit with millisecond time resolution. The performance of this system in activating ChIEF expressed on HEK 293 cells as well as cultured neurons was first evaluated, and the laser stimulation patterns were optimized. Next, the spatiotemporally selective manipulation of multiple neurons was achieved in a precise manner. Finally, we demonstrated the versatility of this high-resolution method in dissecting neural circuits both in the mouse cortical slice and the Drosophila brain in vivo. Taken together, our results show that the combination of AOD-assisted laser stimulation and optogenetic tools provides a flexible solution for manipulating neuronal activity at high efficiency and with high temporal precision

The Role of PPK26 in Drosophila Larval Mechanical Nociception

In Drosophila larvae, the class IV dendritic arborization (da) neurons are polymodal nociceptors. Here, we show that ppk26 (CG8546) plays an important role in mechanical nociception in class IV da neurons. Our immunohistochemical and functional results demonstrate that ppk26 is specifically expressed in class IV da neurons. Larvae with mutant ppk26 showed severe behavioral defects in a mechanical nociception behavioral test but responded to noxious heat stimuli comparably to wild-type larvae. In addition, functional studies suggest that ppk26 and ppk (also called ppk1) function in the same pathway, whereas piezo functions in a parallel pathway. Consistent with these functional results, we found that PPK and PPK26 are interdependent on each other for their cell surface localization. Our work indicates that PPK26 and PPK might form heteromeric DEG/ENaC channels that are essential for mechanotransduction in class IV da neurons

Cleavage cracking of ductile-metal substrates induced by brittle coating fracture

Brittle coatings are often used to protect underlying ductile substrates from damage. Recent experimental observations show that the fracture of a brittle coating can cause the micro-cracking of the ductile metal substrates, threatening the safety and reliability of engineering structures. The cracking mode of the substrates is unclear and the corresponding mechanism remains poorly understood. In the present work, by performing room-temperature uniaxial tension experiments with a 10−4 s−1 straining rate, we have observed cleavage cracking in ductile-metal substrates (pure iron, AISI 1020 steel and brass) coated with WC-10%Co-4%Cr. Theoretical analysis reveals that the cleavage cracking is the result of brittle coating and its fracture, which synergically inhibit the local plastic deformation of the underlying substrate via two mechanisms. One is to restrain the dislocation nucleation and mobility near the interface of the substrate. The other is to bring a local high strain rate loading to the substrate, due to fast crack propagation in the brittle coating. The coupling of two effects leads to the nucleation of cleavage crack in the normally ductile substrates and causes significant loss in their ductility. This detrimental effect will be much more pronounced with the increase in the coating thickness. The findings shed new light on the failure mechanisms of brittle coating-metal substrate and provide guidelines in the material design of such systems.submittedVersionThis is a submitted manuscript of an article published by Elsevier Ltd in Acta Materialia, 11 April 2018

Revealing the microstructures and seepage characteristics in the uncured rubber-cord composites using micro-computed tomography and lattice Boltzmann approach

The internal microstructure distribution of cord-rubber-air during the processing of uncured rubber-cord composites (URCs) determines the finished product's performance. For the first time, we used computed tomography (CT) and the lattice Boltzmann method (LBM) to establish a geometrical representation model of the real microscopic pore-fracture structures of URCs and investigate the seepage law of fluid in porous URCs, where the reinforced rubber formula was originally designed to reduce CT artifacts of cord. The results showed that the average porosity and pore radius of the original cord (0.2711 and 15.53 μm, respectively) were considerably larger than those of the URCs (0.0509 and 4.46 μm, respectively); the pore number of the cord was the largest when the pore radius was 5–10 μm, accounting for 29.36% of the total number; however, the pore number accounted for 31.36% of the total number of the URCs when the pore radius was 2–3 μm. Moreover, the characteristics of the pore/throat surface area and pore volume/throat length exhibited perfect consistent patterns with those of the pore radius. Furthermore, the fluid flow velocity increased in both cord and URCs as the displacement differential pressure increased, but decreased dramatically as the fluid kinematic viscosity increased. The critical values of displacement differential pressure and kinematic viscosity were different in the cord and URCs samples, presenting 11.1209 Pa/1.3696 × 10−3 m2/s and 14.2984 Pa/2.8869 × 10−4 m2/s, respectively. This phenomenon should be attributed that when the uncured rubber was injected into the original cord sample, its porosity decreased, its pore radius decreased, the number of micro-scale pores increased, and flow resistance became larger, resulting in a higher critical value of displacement differential pressure and a lower critical value of kinematic viscosity

Electron cryo-microscopy structure of the mechanotransduction channel NOMPC.

Mechanosensory transduction for senses such as proprioception, touch, balance, acceleration, hearing and pain relies on mechanotransduction channels, which convert mechanical stimuli into electrical signals in specialized sensory cells. How force gates mechanotransduction channels is a central question in the field, for which there are two major models. One is the membrane-tension model: force applied to the membrane generates a change in membrane tension that is sufficient to gate the channel, as in the bacterial MscL channel and certain eukaryotic potassium channels. The other is the tether model: force is transmitted via a tether to gate the channel. The transient receptor potential (TRP) channel NOMPC is important for mechanosensation-related behaviours such as locomotion, touch and sound sensation across different species including Caenorhabditis elegans, Drosophila and zebrafish. NOMPC is the founding member of the TRPN subfamily, and is thought to be gated by tethering of its ankyrin repeat domain to microtubules of the cytoskeleton. Thus, a goal of studying NOMPC is to reveal the underlying mechanism of force-induced gating, which could serve as a paradigm of the tether model. NOMPC fulfils all the criteria that apply to mechanotransduction channels and has 29 ankyrin repeats, the largest number among TRP channels. A key question is how the long ankyrin repeat domain is organized as a tether that can trigger channel gating. Here we present a de novo atomic structure of Drosophila NOMPC determined by single-particle electron cryo-microscopy. Structural analysis suggests that the ankyrin repeat domain of NOMPC resembles a helical spring, suggesting its role of linking mechanical displacement of the cytoskeleton to the opening of the channel. The NOMPC architecture underscores the basis of translating mechanical force into an electrical signal within a cell

Western blotting analysis of SNAT2-HA wild-type and 7 single cysteine mutants modified by NEM and mPEG-Mal implicates Cys245 and Cys279 in a disulfide bridge.

<p>Lysates of HEK293T cells expressing either wild-type or 7 single cysteine mutants were processed by sequential treatment of NEM, DTT, and mPEG-Mal under denatured condition. Controls were treated without DTT. The proteins were separated on an 8% SDS gel, detected by immunodetection with the mouse anti-HA tag antibody followed by the HRP-conjugated goat anti-mouse secondary antibody. Anti-GAPDH mouse monoclonal antibody was used to detect the GAPDH as an internal reference.</p

Phospholipid Homeostasis Regulates Dendrite Morphogenesis in Drosophila Sensory Neurons

Summary: Disruptions in lipid homeostasis have been observed in many neurodevelopmental disorders that are associated with dendrite morphogenesis defects. However, the molecular mechanisms of how lipid homeostasis affects dendrite morphogenesis are unclear. We find that easily shocked (eas), which encodes a kinase with a critical role in phospholipid phosphatidylethanolamine (PE) synthesis, and two other enzymes in this synthesis pathway are required cell autonomously in sensory neurons for dendrite growth and stability. Furthermore, we show that the level of Sterol Regulatory Element-Binding Protein (SREBP) activity is important for dendrite development. SREBP activity increases in eas mutants, and decreasing the level of SREBP and its transcriptional targets in eas mutants largely suppresses the dendrite growth defects. Furthermore, reducing Ca2+ influx in neurons of eas mutants ameliorates the dendrite morphogenesis defects. Our study uncovers a role for EAS kinase and reveals the in vivo function of phospholipid homeostasis in dendrite morphogenesis. : Meltzer et al. show that EAS, a conserved kinase in the phospholipid phosphatidylethanolamine synthesis pathway, regulates dendrite growth via SREBP signaling and Ca2+ influx. Their study reveals the role of phospholipid homeostasis in dendrite morphogenesis in vivo. Keywords: dendrite, sensory neurons, morphogenesis, Drosophila, phospholipid, homeostasis, lipi