Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-honmachi, Sakyo-ku, Kyoto, Japan.
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-honmachi, Sakyo-ku, Kyoto, Japan; Laboratory of Pharmacology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, Japan.
Brain Res Bull. 2021 Aug;173:45-52. doi: 10.1016/j.brainresbull.2021.05.003. Epub 2021 May 12.
Cerebral infarct is caused by cerebrovascular occlusion and results in brain damage. Although many rodent models of cerebral infarct exist, there is none based on zebrafish. In this study, we developed a novel ischemia-reperfusion model induced by hypoxic treatment using zebrafish. We first examined the changes in blood flow under hypoxic conditions. Hypoxic treatment interrupted the blood flow in 4 dpf (days post fertilization) zebrafish larvae. To quantify the trunk and cerebral blood flow, we selected the middle mesencephalic central artery (MMCtA) as a cerebral blood vessel and the dorsal aorta (DA) as a blood vessel of the trunk. Interestingly, the interruption of blood flow in MMCtA preceded that in DA. Considering these results, we hypothesized that reoxygenation immediately after hypoxia-induced cerebral ischemia leads to reperfusion. As a result, hypoxia-reoxygenation (H/R) treatment induced ischemia-reperfusion in cerebral vessels. Furthermore, brain cell death was increased 24 h after H/R treatment. Transgenic zebrafish (HuC:kaede), with neuronal cells expressing the kaede fluorescent protein, was used to investigate the effect of H/R on neuronal cells. The H/R treatment reduced the fluorescence intensity of kaede. Besides, glial fibrillary acidic protein immunoreactivity in H/R-treated larvae was significantly increased. In conclusion, H/R-treated zebrafish larvae may provide a novel ischemia-reperfusion model.
脑梗死是由脑血管阻塞引起的，导致脑损伤。虽然有许多啮齿动物脑梗死模型，但没有基于斑马鱼的模型。在这项研究中，我们开发了一种使用斑马鱼的新的缺血再灌注模型，通过缺氧处理。我们首先检查了缺氧条件下血流的变化。缺氧处理中断了 4 dpf（受精后天数）斑马鱼幼虫的血流。为了量化躯干和脑血流，我们选择中脑中央动脉（MMCtA）作为脑血管，背主动脉（DA）作为躯干血管。有趣的是，MMCtA 中的血流中断先于 DA 中的血流中断。考虑到这些结果，我们假设缺氧诱导的脑缺血后立即再氧合导致再灌注。结果，缺氧-复氧（H/R）处理诱导了脑血管的缺血再灌注。此外，H/R 处理后 24 小时脑细胞死亡增加。转基因为神经元细胞表达 kaede 荧光蛋白的斑马鱼（HuC:kaede）用于研究 H/R 对神经元细胞的影响。H/R 处理降低了 kaede 的荧光强度。此外，H/R 处理幼虫中的胶质纤维酸性蛋白免疫反应性显著增加。总之，H/R 处理的斑马鱼幼虫可能提供了一种新的缺血再灌注模型。
