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苯硫脲介导的实验性脱色可减少斑马鱼幼虫戊四氮致痫反应。

Phenylthiourea-mediated experimental depigmentation reduces seizurogenic response of pentylenetetrazol in zebrafish larva.

机构信息

Pharmacology and Toxicology Laboratory, Dietetics and Nutrition Technology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur 176061, Himachal Pradesh, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India.

Pharmacology and Toxicology Laboratory, Dietetics and Nutrition Technology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur 176061, Himachal Pradesh, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India.

出版信息

J Pharmacol Toxicol Methods. 2024 Jul-Aug;128:107532. doi: 10.1016/j.vascn.2024.107532. Epub 2024 Jun 7.

Abstract

Zebrafish larvae exposed to chemoconvulsants show behavioral seizures and electrographic abnormalities similar to the other mammalian models, making it a potential tool in epilepsy research. During the embryonic stage, zebrafish remains transparent which enables real-time developmental detection and in-situ gene/protein expression. However, pigmentation during the larval stage restricts transparency. Phenylthiourea (1-phenyl-2-thiourea; PTU) is a commonly used pigmentation blocker that maintains larval transparency. It is widely used along with chemoconvulsants to study in situ expressions in epileptic larvae, however, its effect on seizures largely remains unknown. Therefore, in the present study, the effect of PTU-mediated depigmentation was studied on pentylenetetrazol (PTZ)-induced seizures in zebrafish larvae. After spawning, the fish embryos were subjected to standard depigmentation protocol using 0.13 mM PTU. At 7-days post fertilization seizures were induced using 8 mM PTZ. PTU exposure significantly reduced PTZ-mediated hyperactive responses indicated by decreased distance travelled and swimming velocity of the larvae. Furthermore, PTU-exposed depigmented larvae also showed an increase in the latency to the onset of PTZ-mediated clonic-like seizures. The results concluded that PTU depigmentation protocol reduces the seizurogenic response of PTZ, hence its usage for imaging zebrafish larvae must be carefully monitored to avoid erroneous results.

摘要

斑马鱼幼虫暴露于化学惊厥剂中会表现出类似于其他哺乳动物模型的行为性惊厥和脑电图异常,使其成为癫痫研究的潜在工具。在胚胎期,斑马鱼仍然是透明的,这使得能够进行实时发育检测和原位基因/蛋白质表达。然而,幼虫期的色素沉着会限制透明度。苯硫脲(1-苯基-2-硫脲;PTU)是一种常用的色素沉着抑制剂,可保持幼虫的透明度。它与化学惊厥剂一起广泛用于研究癫痫幼虫的原位表达,但它对惊厥的影响在很大程度上仍然未知。因此,在本研究中,研究了 PTU 介导的脱色素作用对斑马鱼幼虫戊四氮(PTZ)诱导的惊厥的影响。产卵后,使用 0.13 mM PTU 对鱼胚胎进行标准脱色素处理。在受精后 7 天,使用 8 mM PTZ 诱导惊厥。PTU 暴露显著降低了 PTZ 介导的过度活跃反应,表现为幼虫游动距离和游泳速度降低。此外,PTU 暴露的脱色素幼虫在 PTZ 诱导的类似阵挛性惊厥发作的潜伏期也增加。结果表明,PTU 脱色素方案降低了 PTZ 的致惊厥反应,因此在使用其对斑马鱼幼虫进行成像时必须仔细监测,以避免错误的结果。

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