Vimentin and 70K Neurofilament Protein Co-exist in Embryonic Neurones from Spinal Ganglia

Abstract: The mesenchymal intermediate filament protein vimentin and the 70K component of neurofilament were detected by two-dimensional gel electrophoresis in cultures of pure sensory and sympathetic neurones derived from chick embryos. The identities of these neuronal intermediate filament proteins were confirmed by comparison of their molecular weights, isoelectric points, and peptide patterns from limited papain digestions with those of the corresponding proteins from fibroblasts and brain, respectively. A specific antibody to vimentin stained filamenteous structures and colcemid-induced coils in both neurones and associated satellite cells. In contrast, a specific antibody to the 70K neurofilament protein stained these structures solely in neurones. This neurone-specific staining, as well as its molecular weight and isoelectric point, distinguishes the 70K neurofilament protein from the 68K neurofilament as sociated protein described by others, which has been claimed to resemble the tubulin assembly protein.     

The ultrastructure of the so-called olfactory setae on the antennula ofDaphnia magna Straus (Crustacea,Cladocera)

A group of nine sensory setae is found on the tip of the antennula ofDaphnia magna in both sexes. Inside a seta four dendrites are situated, each with one receptor cilium. The receptor cilia extend through a liquor space into the exterior part of the seta. The exterior part of the liquor space is divided from the interior part by a knob-like thickening of the innermost layer of the epicuticle, the basal bead. The basal bead narrows the liquor space and the receptor cilia. The interior part of the liquor space is surrounded by five sheath cells, the exterior part by a thin cuticle. In the exterior part the receptor cilia branch partly and reach a terminal pellet on the tip of the seta. The terminal pellet is a thickened part of the epicuticle. It is permeable to several dissolved substances. It is the exterior part of the receptor that projects over the tip of the antennula and seems to be the entire seta. During the premoult the fifth sheath cell builds up the articulation of the seta, the fourth the basal bead, and the third the shaft of the seta. The first sheath cell forms the cuticular sheath. The organ seems to be a chemoreceptor, but the adequate stimulus is as yet unknown.     

Resonance Raman and FTIR spectroscopic characterization of the closed and open states of channelrhodopsin-1

Channelrhodopsin-1 from Chlamydomonas augustae (CaChR1) is a light-activated cation channel, which is a promising optogenetic tool. We show by resonance Raman spectroscopy and retinal extraction followed by high pressure liquid chromatography (HPLC) that the isomeric ratio of all-trans to 13-cis of solubilized channelrhodopsin-1 is with 70:30 identical to channelrhodopsin-2 from Chlamydomonas reinhardtii (CrChR2). Critical frequency shifts in the retinal vibrations are identified in the Raman spectrum upon transition to the open (conductive P₂³⁸⁰) state. Fourier transform infrared spectroscopy (FTIR) spectra indicate different structures of the open states in the two channelrhodopsins as reflected by the amide I bands and the protonation pattern of acidic amino acids.     

Freezing behaviour facilitates bioelectric crypsis in cuttlefish faced with predation risk

Cephalopods, and in particular the cuttlefish Sepia officinalis, are common models for studies of camouflage and predator avoidance behaviour. Preventing detection by predators is especially important to this group of animals, most of which are soft-bodied, lack physical defences, and are subject to both visually and non-visually mediated detection. Here, we report a novel cryptic mechanism in S. officinalis in which bioelectric cues are reduced via a behavioural freeze response to a predator stimulus. The reduction of bioelectric fields created by the freeze-simulating stimulus resulted in a possible decrease in shark predation risk by reducing detectability. The freeze response may also facilitate other non-visual cryptic mechanisms to lower predation risk from a wide range of predator types.     

Novel Immunohistochemical Techniques Using Discrete Signal Amplification Systems for Human Cutaneous Peripheral Nerve Fiber Imaging

Confocal imaging uses immunohistochemical binding of specific antibodies to visualize tissues, but technical obstacles limit more widespread use of this technique in the imaging of peripheral nerve tissue. These obstacles include same-species antibody cross-reactivity and weak fluorescent signals of individual and co-localized antigens. The aims of this study were to develop new immunohistochemical techniques for imaging of peripheral nerve fibers. Three-millimeter punch skin biopsies of healthy individuals were fixed, frozen, and cut into 50-µm sections. Tissues were stained with a variety of antibody combinations with two signal amplification systems, streptavidin-biotin-fluorochrome (sABC) and tyramide-horseradish peroxidase-fluorochrome (TSA), used simultaneously to augment immunohistochemical signals. The combination of the TSA and sABC amplification systems provided the first successful co-localization of sympathetic adrenergic and sympathetic cholinergic nerve fibers in cutaneous human sweat glands and vasomotor and pilomotor systems. Primary antibodies from the same species were amplified individually without cross-reactivity or elevated background interference. The confocal fluorescent signal-to-noise ratio increased, and image clarity improved. These modifications to signal amplification systems have the potential for widespread use in the study of human neural tissues.     

Genetic aberrations in macroautophagy genes leading to diseases

The catabolic process of macroautophagy, through the rapid degradation of unwanted cellular components, is involved in a multitude of cellular and organismal functions that are essential to maintain homeostasis. Those functions include adaptation to starvation, cell development and differentiation, innate and adaptive immunity, tumor suppression, autophagic cell death, and maintenance of stem cell stemness. Not surprisingly, an impairment or block of macroautophagy can lead to severe pathologies. A still increasing number of reports, in particular, have revealed that mutations in the autophagy-related (ATG) genes, encoding the key players of macroautophagy, are either the cause or represent a risk factor for the development of several illnesses. The aim of this review is to provide a comprehensive overview of the diseases and disorders currently known that are or could be caused by mutations in core ATG proteins but also in the so-called autophagy receptors, which provide specificity to the process of macroautophagy. Our compendium underlines the medical relevance of this pathway and underscores the importance of the eventual development of therapeutic approaches aimed at modulating macroautophagy.     

Mapping protein interactions of sodium channel NaV1.7 using epitope‐tagged gene‐targeted mice

The voltage‐gated sodium channel Na_V1.7 plays a critical role in pain pathways. We generated an epitope‐tagged Na_V1.7 mouse that showed normal pain behaviours to identify channel‐interacting proteins. Analysis of Na_V1.7 complexes affinity‐purified under native conditions by mass spectrometry revealed 267 proteins associated with Nav1.7 in vivo. The sodium channel β3 (Scn3b), rather than the β1 subunit, complexes with Nav1.7, and we demonstrate an interaction between collapsing‐response mediator protein (Crmp2) and Nav1.7, through which the analgesic drug lacosamide regulates Nav1.7 current density. Novel Na_V1.7 protein interactors including membrane‐trafficking protein synaptotagmin‐2 (Syt2), L‐type amino acid transporter 1 (Lat1) and transmembrane P24‐trafficking protein 10 (Tmed10) together with Scn3b and Crmp2 were validated by co‐immunoprecipitation (Co‐IP) from sensory neuron extract. Nav1.7, known to regulate opioid receptor efficacy, interacts with the G protein‐regulated inducer of neurite outgrowth (Gprin1), an opioid receptor‐binding protein, demonstrating a physical and functional link between Nav1.7 and opioid signalling. Further information on physiological interactions provided with this normal epitope‐tagged mouse should provide useful insights into the many functions now associated with the Na_V1.7 channel.     

微血管减压术联合感觉根部分切断术对原发性三叉神经痛患者疼痛评分、生活质量及睡眠状况的影响

目的:探讨微血管减压术(MVD)联合感觉根部分切断术(PSR)对原发性三叉神经痛(TN)患者疼痛评分、生活质量及睡眠状况的影响。方法:回顾性分析2015年2月~2019年3月期间我院收治的80例原发性TN患者的临床资料,根据手术方式的不同将患者分为对照组(n=40,MVD治疗)和研究组(n=40,MVD联合PSR治疗),比较两组患者疼痛评分、生活质量、围术期指标、睡眠状况、并发症发生情况以及复发率。结果:两组患者治疗后视觉疼痛模拟量表(VAS)评分均较治疗前下降,且研究组低于对照组(P<0.05)。两组患者治疗后生活质量量表(SF-36)各维度评分均较治疗前升高,且研究组高于对照组(P<0.05)。两组患者治疗后匹兹堡睡眠质量指数表(PSQI)各项目评分均较治疗前升高,且研究组高于对照组(P<0.05)。研究组住院时间短于对照组,手术时间长于对照组(P<0.05);两组术中出血量比较无统计学差异(P>0.05)。研究组的并发症总发生率低于对照组(P<0.05)。两组随访期间复发率比较差异无统计学意义(P>0.05)。结论:MVD联合PSR治疗原发性TN,虽然手术时间较长,但是在减轻患者疼痛、改善患者生活质量及睡眠状况等方面效果显著,能够降低并发症发生率。     

Foraging costs drive female resistance to a sensory trap

Male ornaments can evolve through the exploitation of female perceptual biases such as those involved in responding to cues from food. This type of sensory exploitation may lead to confusion between the male signals and the cues that females use to find/recognize food. Such interference would be costly to females and may be one reason why females evolve resistance to the male ornaments. Using a group of species of viviparous fish where resistance to a sensory trap has evolved, we demonstrate that females exposed to an ornament that resembles food have a diminished foraging efficiency, that this effect is apparent when foraging on a food item with which the ornament shares visual attributes, and that not all species are equally affected by such confusion. Our results lend support to the model of ornamental evolution through chase-away sexual conflict.