Department of Plant and Animal Biology, Faculty of Biological Sciences and Technology, University of Isfahan, Isfahan, Iran.
Department of Physiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Protein J. 2023 Oct;42(5):586-595. doi: 10.1007/s10930-023-10140-6. Epub 2023 Aug 2.
Spider venom contains various peptides and proteins, which can be used for pharmacological applications. Finding novel therapeutic strategies against neurodegenerative diseases with the use of purified peptides and proteins, extracted from spiders can be greatly precious. Neurodegenerative diseases are rapidly developing and expanding all over the world. Excitotoxicity is a frequent condition amongst neuro-degenerative disorders. This harmful process is usually induced through hyper-activation of N-Methyl-D-Aspartate (NMDA) receptor, and P/Q-type voltage-gated calcium channels (VGCCs). The omega-agatoxin-Aa4b is a selective and strong VGCCblocker. This study aimed to investigate the effects of this blocker on the NMDA-induced memory and learning defect in rats. For this purpose, nineteen spiders of the funnel-weaver Agelena orientalis species were collected. The extracted venom was lyophilized andpurified through gel-filtration chromatography, and capillary electrophoresis techniques. Subsequently, mass spectrometry (HPLC-ESI-MS) was used for identification of this bio-active small protein. Afterward, the effect of the omega-agatoxin-Aa4b (2 μg, intra-cornu ammonis-3 of the hippocampus) on the NMDA-induced learning and memory deficits in rats was evaluated. Learning and memory performances were evaluated by the use of passive avoidance test. For synaptic quantification and memory function the amount of calcium/calmodulin-dependent protein kinase ІІ (CaCdPKІІ) gene expression was measured using the Real-time PCR technique. To compare the experimental groups, hematoxylin and eosin (H&E) staining of hippocampus tissues was performed. Our results rendered that the omega-Agatoxin-Aa4b treatment can ameliorate and reverse the learning and memory impairment caused by NMDA-induced excitotoxicity in rat hippocampus.
蜘蛛毒液含有各种肽和蛋白质,可用于药理学应用。使用从蜘蛛中提取的纯化肽和蛋白质寻找针对神经退行性疾病的新型治疗策略非常宝贵。神经退行性疾病在全球范围内迅速发展和扩大。兴奋性毒性是神经退行性疾病中的常见情况。这种有害过程通常是通过 N-甲基-D-天冬氨酸(NMDA)受体和 P/Q 型电压门控钙通道(VGCC)的过度激活引起的。ω-芋螺毒素-Aa4b 是一种选择性和强 VGCC 阻断剂。本研究旨在研究该阻断剂对大鼠 NMDA 诱导的记忆和学习缺陷的影响。为此,收集了 19 只漏斗蛛属 Agelena orientalis 种的蜘蛛。提取的毒液经冻干后,通过凝胶过滤层析和毛细管电泳技术进行纯化。随后,使用质谱(HPLC-ESI-MS)鉴定这种生物活性小蛋白。之后,评估了ω-芋螺毒素-Aa4b(2μg,海马角 3 区)对 NMDA 诱导的大鼠学习和记忆缺陷的影响。使用被动回避试验评估学习和记忆性能。为了进行突触定量和记忆功能分析,使用实时 PCR 技术测量钙/钙调蛋白依赖性蛋白激酶Ⅱ(CaCdPKⅡ)基因表达的量。为了比较实验组,对海马组织进行了苏木精和伊红(H&E)染色。我们的结果表明,ω-芋螺毒素-Aa4b 处理可以改善和逆转 NMDA 诱导的兴奋性毒性引起的大鼠海马学习和记忆障碍。