Huang Xing, Wang Chuang, Li Song, Liu Yubo, Zhang Zhichao
Department of Radiolgy, Jilin Province People's Hospital, Jilin, 130021, People's Republic of China.
J Fluoresc. 2014 Jul;24(4):1313-7. doi: 10.1007/s10895-014-1416-x. Epub 2014 Jun 10.
In the present study, we used the previously developed fluorescence probe, EPNP, to generate the first image of the distribution of mercuric ion in primary mouse neuron cultures. At postnatal day 1 (P1), the mice were intraperitoneally (IP) injected with mercuric chloride in doses ranging from 0.05 to 0.6 μg/g body weight. After 1, 2, 3, and 4 days exposure, primary nervous cell cultures and frozen brain and spinal tissue sections were prepared and dyed using EPNP. On the third day of repeated injections, Hg(2+) was visualized in primary cerebral neuron cultures as an increase of Hg(2+)-induced fluorescence at the doses ≥ 0.1 μg/g. A similar accumulation of Hg(2+) was observed in frozen hippocampus tissue sections. In contrast, no Hg(2+) was observed in spinal cord neurons and spinal tissue sections. The detection of a low dose of IP injected mercury in mouse cerebral neurons facilitated the evaluation of the exposure risk to low-dose Hg(2+) in immature organisms. Moreover, the highly sensitive EPNP revealed Hg(2+) in the cerebral neurons of mice younger than P4, while the presence of Hg(2+) was not detected until ≥ P11 in previous reports. Thus, this technology and the results obtained herein are of interest for neurotoxicology.
在本研究中,我们使用先前开发的荧光探针EPNP,生成了原代小鼠神经元培养物中汞离子分布的首张图像。在出生后第1天(P1),给小鼠腹腔注射剂量范围为0.05至0.6μg/g体重的氯化汞。暴露1、2、3和4天后,制备原代神经细胞培养物以及冷冻的脑和脊髓组织切片,并用EPNP进行染色。在重复注射的第三天,在原代脑神经元培养物中,当剂量≥0.1μg/g时,Hg(2+)以Hg(2+)诱导的荧光增加的形式显现出来。在冷冻的海马组织切片中也观察到类似的Hg(2+)积累。相比之下,在脊髓神经元和脊髓组织切片中未观察到Hg(2+)。在小鼠脑神经元中检测到腹腔注射的低剂量汞,有助于评估未成熟生物体中低剂量Hg(2+)的暴露风险。此外,高灵敏度的EPNP在小于P4的小鼠脑神经元中检测到了Hg(2+),而在先前的报告中,直到≥P11才检测到Hg(2+)的存在。因此,这项技术以及本文获得的结果对神经毒理学具有重要意义。