Gustatory sensitivity of the external taste buds of Oreochromis niloticus L. to amino acids

The gustatory sensitivity of the Nile tilapia, Oreochromis niloticus L., to different amino acids was studied using an electrophysiological approach. The electrical responses were recorded from a branch of the facial nerve innervating the external taste buds of the upper lip. The relative stimulatory effectiveness (RSE) of nine amino acids and betaine were determined at a concentration of 1 mm and all of them elicited neural responses. This species responded well to the neutral, basic and acidic amino acids. The most stimulatory amino acids were L‐histidine, L‐arginine, L‐serine, L‐methionine and L‐glutamine; L‐proline and betaine were the least stimulatory. The results of this study suggest that the Nile tilapia has high external gustatory sensitivity to some amino acids as a physiological adaptation to search effectively for their sources. The effect of the pH, ranging from 4.0 to 9.0, on the RSE of three neutral amino acids and artificial pond water (APW) was also studied. The RSE increased below pH 6.0 and was relatively unaffected from 7.0 to 9.0, indicating that acidified stimulants are highly stimulatory in this species. Nile tilapia did not discriminate the pH of APW as effectively as some of the species studied earlier.     

苦参碱对低钾致兔左心室流出道细胞电生理异常保护作用及其机制

目的探讨苦参碱对低钾致兔左心室流出道细胞电生理异常保护作用及其机制.方法应用常规玻璃微电极细胞内记录技术,观察正常灌流液、低钾灌流液、低钾＋苦参碱（50μmol/L）灌流液对兔左心室流出道慢反应自律细胞最大舒张电位（MDP）,动作电位0相幅值（APA）,50%复极化时间（APD50）,90%复极化时间（APD90）,4...     

Ligands of the nicotinic acetylcholine receptor as insecticides

Insect nicotinic acetyl receptors (nAChR) are targets of growing importance and, since the early 1990s, the number of such highly effective insecticides as imidacloprid and spinosyn has grown. Several natural compounds, eg dihydro-β-erythroidine, methyl caconitine and paraherquamide, showing high affinity to the same receptor, were considerably less active as insecticides, most likely because of their antagonistic action. Our observations on aphids after ingestion of the antagonistic compound dihydro-β-erythroidine revealed anti-feedant-like properties. As a consequence, the symptomology of poisoning was totally different between agonists and antagonists of the nAChR. Electrophysiological (whole-cell voltage clamp) measurements in isolated housefly neurones revealed that agonism seems to be a prerequisite for insecticidal activity. Furthermore, we were able to demonstrate the existence of two different subtypes of the nAChR in isolated locust neurones with different pharmacology and ion-channel properties.     

The innervation pattern of fast myotomal muscle in the teleostMyoxocephalus scorpius: A reappraisal

The pattern of polyneuronal innervation in the sculpinMyoxocephalus scorpius was examined. The majority of fast fibres in sculpin are innervated by both adjacent nerves, and <15% receive input from sub-adjacent myotomes. Stimulation through both adjacent and sub-adjacent nerves results in muscle action potentials. Each adjacent nerve supplies 2–5 axons to a given fibre. A typical fibre is probably innervated by around 4–6 axons, and pre-terminal branching accounts for the 8–20 endplates present on these fibres.     

Fundamentals of arrhythmogenic mechanisms and treatment strategies for equine atrial fibrillation

Atrial fibrillation (AF) is the most common pathological arrhythmia in horses. Although it is not usually a life-threatening condition on its own, it can cause poor performance and make the horse unsafe to ride. It is a complex multifactorial disease influenced by both genetic and environmental factors including exercise training, comorbidities or ageing. The interactions between all these factors in horses are still not completely understood and the pathophysiology of AF remains poorly defined. Exciting progress has been recently made in equine cardiac electrophysiology in terms of diagnosis and documentation methods such as cardiac mapping, implantable electrocardiogram (ECG) recording devices or computer-based ECG analysis that will hopefully improve our understanding of this disease. The available pharmaceutical and electrophysiological treatments have good efficacy and lead to a good prognosis for AF, but recurrence is a frequent issue that veterinarians have to face. This review aims to summarise our current understanding of equine cardiac electrophysiology and pathophysiology of equine AF while providing an overview of the mechanism of action for currently available treatments for equine AF.     

Synthesis,Structural and Pharmacological Characterizations of CIC,a Novel α-Conotoxin with an Extended N-Terminal Tail

Cone snails are venomous marine predators that rely on fast-acting venom to subdue their prey and defend against aggressors. The conotoxins produced in the venom gland are small disulfide-rich peptides with high affinity and selectivity for their pharmacological targets. A dominant group comprises α-conotoxins, targeting nicotinic acetylcholine receptors. Here, we report on the synthesis, structure determination and biological activity of a novel α-conotoxin, CIC, found in the predatory venom of the piscivorous species Conus catus and its truncated mutant Δ-CIC. CIC is a 4/7 α-conotoxin with an unusual extended N-terminal tail. High-resolution NMR spectroscopy shows a major influence of the N-terminal tail on the apparent rigidity of the three-dimensional structure of CIC compared to the more flexible Δ-CIC. Surprisingly, this effect on the structure does not alter the biological activity, since both peptides selectively inhibit α3β2 and α6/α3β2β3 nAChRs with almost identical sub- to low micromolar inhibition constants. Our results suggest that the N-terminal part of α-conotoxins can accommodate chemical modifications without affecting their pharmacology.     

��-Conotoxin GVIA Mimetics that Bind and Inhibit Neuronal Cav2.2 Ion Channels

The neuronal voltage-gated N-type calcium channel (Ca_v2.2) is a validated target for the treatment of neuropathic pain. A small library of anthranilamide-derived ω-Conotoxin GVIA mimetics bearing the diphenylmethylpiperazine moiety were prepared and tested using three experimental measures of calcium channel blockade. These consisted of a ¹²⁵I-ω-conotoxin GVIA displacement assay, a fluorescence-based calcium response assay with SH-SY5Y neuroblastoma cells, and a whole-cell patch clamp electrophysiology assay with HEK293 cells stably expressing human Ca_v2.2 channels. A subset of compounds were active in all three assays. This is the first time that compounds designed to be mimics of ω-conotoxin GVIA and found to be active in the ¹²⁵I-ω-conotoxin GVIA displacement assay have also been shown to block functional ion channels in a dose-dependent manner.     

Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli

α-Conotoxin LvIA is derived from Conus lividus, native to Hainan, and is the most selective inhibitor of α3β2 nicotinic acetylcholine receptors (nAChRs) known to date. In this study, an efficient approach for the production of recombinant α-Conotoxin LvIA is described. Tandem repeats of a LvIA gene fragment were constructed and fused with a KSI gene and a His₆ tag in a Escherichia coli (E. coli) expression vector pET-31b(+). The recombinant plasmids were transformed into E. coli and were found to express well. The KSI-(LvIA)_n-His₆ fusion protein was purified by metal affinity chromatography and then cleaved with CNBr to release recombinant LvIA (rLvIA). High yields of fusion protein ranging from 100 to 500 mg/L culture were obtained. The pharmacological profile of rLvIA was determined by two-electrode voltage-clamp electrophysiology in Xenopus laevis oocytes expressing rat nAChR subtypes. The rLvIA antagonized the α3β2 nAChR subtype selectively with a nano-molar IC₅₀. The rLvIA was analgesic in a mouse hot-plate test model of pain. Overall, this study provides an effective method to synthesize α-conotoxin LvIA in an E. coli recombinant expression system, and this approach could be useful to obtain active conopeptides in large quantity and at low cost.