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利用双光子显微镜进行钙成像,以检测施用以新烟碱类杀虫剂的小鼠大脑功能的破坏。

Ca imaging with two-photon microscopy to detect the disruption of brain function in mice administered neonicotinoid insecticides.

机构信息

Laboratory of Toxicology, Department of Environmental Veterinary Sciences, Faculty of Veterinary Medicine, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo, 060-0818, Japan.

Department of Anatomy and Molecular Cell Biology, Nagoya University Graduate School of Medicine, 65 Tsurumi-cho, Showa-ku, Nagoya, 466-8550, Japan.

出版信息

Sci Rep. 2022 Mar 24;12(1):5114. doi: 10.1038/s41598-022-09038-7.

Abstract

Neonicotinoid pesticides are a class of insecticides that reportedly have harmful effects on bees and dragonflies, causing a reduction in their numbers. Neonicotinoids act as neuroreceptor modulators, and some studies have reported their association with neurodevelopmental disorders. However, the precise effect of neonicotinoids on the central nervous system has not yet been identified. Herein, we conducted in vivo Ca imaging using a two-photon microscope to detect the abnormal activity of neuronal circuits in the brain after neonicotinoid application. The oral administration of acetamiprid (ACE) (20 mg/kg body weight (BW) in mature mice with a quantity less than the no-observed-adverse-effect level (NOAEL) and a tenth or half of the median lethal dose (LD) of nicotine (0.33 or 1.65 mg/kg BW, respectively), as a typical nicotinic acetylcholine receptor (nAChR) agonist, increased anxiety-like behavior associated with altered activities of the neuronal population in the somatosensory cortex. Furthermore, we detected ACE and its metabolites in the brain, 1 h after ACE administration. The results suggested that in vivo Ca imaging using a two-photon microscope enabled the highly sensitive detection of neurotoxicant-mediated brain disturbance of nerves.

摘要

新烟碱类杀虫剂是一类杀虫剂,据报道对蜜蜂和蜻蜓有有害影响,导致它们的数量减少。新烟碱类杀虫剂作为神经受体调节剂,一些研究报告称它们与神经发育障碍有关。然而,新烟碱类对中枢神经系统的确切影响尚未确定。在此,我们使用双光子显微镜进行体内 Ca 成像,以检测新烟碱类应用后大脑神经元回路的异常活动。在成熟的老鼠中,口服乙酰甲胺磷(ACE)(20mg/kg 体重(BW),低于无观察到不良效应水平(NOAEL),并且是尼古丁(0.33 或 1.65mg/kg BW,分别为)的十分之一或一半,作为典型的烟碱型乙酰胆碱受体(nAChR)激动剂,增加了与躯体感觉皮层神经元群体活动改变相关的焦虑样行为。此外,我们在 ACE 给药 1 小时后检测到大脑中的 ACE 及其代谢物。结果表明,使用双光子显微镜进行的体内 Ca 成像能够高度敏感地检测神经毒剂介导的神经对大脑的干扰。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d964/8948258/58211f940d71/41598_2022_9038_Fig1_HTML.jpg

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