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Purified Native Protein Extracted from the Venom of Agelena orientalis Attenuates Memory Defects in the Rat Model of Glutamate-Induced Excitotoxicity

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Abstract

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.

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Data Availability

The data that support the findings of this study are available on reasonable request from the corresponding author.

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Acknowledgements

All experimental procedures were approved by the Animal Ethics Committee of the University of Isfahan, and complied with ARRIVE guidelines. In addition, all methods were carried out in accordance with relevant guidelines and regulations like National Research Council’s Guide for the Care and Use of Laboratory Animals. We thank to Najmeh Ghorbani (laboratory expert, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran) for her kind assistance.

Funding

This work was supported by the Department of Animal and Plant Biology, Faculty of Biological Sciences and Technology, University of Isfahan. Isfahan, Iran (Grant No: 154380).

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Author notes

  1. Mohammad Keimasi and Kowsar Salehifard share first authorship.

Authors and Affiliations

  1. Department of Plant and Animal Biology, Faculty of Biological Sciences and Technology, University of Isfahan, Isfahan, Iran

    Mohammad Keimasi, Sayyed Jafar Hoseini, Fariba Esmaeili, Noushin Mirshah Jafar Esfahani, Mohammadjavad Keimasi & Majid Moradmand

  2. Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

    Mohammad Reza Mofid

  3. Department of Physiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    Kowsar Salehifard

  4. Department of Clinical Pharmacy and Pharmacy Practice, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

    Mohammadreza Amirsadri

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Contributions

MK conceived the original idea. MRM, MM, and MK planned the experiments. MK, KS, SJH, FE, NMJE, and MJK performed the experiments, data collection, analysis, and interpretation. MK and MRA wrote the manuscript. All authors revised manuscript. MRM, MM, and MRA have collaborated in presenting the research idea. MRM, MM supervised the project. All authors approved the final version of manuscript.

Corresponding authors

Correspondence to Majid Moradmand or Mohammad Reza Mofid.

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Keimasi, M., Salehifard, K., Hoseini, S.J. et al. Purified Native Protein Extracted from the Venom of Agelena orientalis Attenuates Memory Defects in the Rat Model of Glutamate-Induced Excitotoxicity. Protein J 42, 586–595 (2023). https://doi.org/10.1007/s10930-023-10140-6

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