College of Medicine and Public Health, Flinders University, P.O. Box 2100, Adelaide, SA 5001, Australia.
Flinders Health and Medical Research Institute, Flinders University, Adelaide, SA 5001, Australia.
Viruses. 2024 Aug 20;16(8):1330. doi: 10.3390/v16081330.
Zika virus (ZIKV) impacts the developing brain. Here, a technique was applied to define, in 3D, developmental changes in the brains of ZIKV-infected mice. Postnatal day 1 mice were uninfected or ZIKV-infected, then analysed by iodine staining and micro-CT scanning (diffusible iodine contrast-enhanced micro-CT; diceCT) at 3-, 6-, and 10-days post-infection (dpi). Multiple brain regions were visualised using diceCT: the olfactory bulb, cerebrum, hippocampus, midbrain, interbrain, and cerebellum, along with the lens and retina of the eye. Brain regions were computationally segmented and quantitated, with increased brain volumes and developmental time in uninfected mice. Conversely, in ZIKV-infected mice, no quantitative differences were seen at 3 or 6 dpi when there were no clinical signs, but qualitatively, diverse visual defects were identified at 6-10 dpi. By 10 dpi, ZIKV-infected mice had significantly lower body weight and reduced volume of brain regions compared to 10 dpi-uninfected or 6 dpi ZIKV-infected mice. Nissl and immunofluorescent Iba1 staining on post-diceCT tissue were successful, but RNA extraction was not. Thus, diceCT shows utility for detecting both 3D qualitative and quantitative changes in the developing brain of ZIKV-infected mice, with the benefit, post-diceCT, of retaining the ability to apply traditional histology and immunofluorescent analysis to tissue.
寨卡病毒（ZIKV）会影响发育中的大脑。在这里，应用一种技术来定义 ZIKV 感染小鼠大脑的三维发育变化。在感染后 1 天的新生小鼠中，将未感染或感染 ZIKV 的小鼠进行碘染色和微 CT 扫描（弥散碘对比增强微 CT；diceCT），分别在感染后 3、6 和 10 天（dpi）进行分析。使用 diceCT 可以可视化多个大脑区域：嗅球、大脑、海马体、中脑、间脑和小脑，以及眼睛的晶状体和视网膜。通过计算分割和定量大脑区域，未感染的小鼠的大脑体积增加，且随着发育时间的延长而增加。相反，在 ZIKV 感染的小鼠中，在没有临床症状的 3 或 6 dpi 时，没有观察到定量差异，但在 6-10 dpi 时，定性地发现了不同的视觉缺陷。在 10 dpi 时，与 10 dpi 未感染或 6 dpi ZIKV 感染的小鼠相比，ZIKV 感染的小鼠的体重显著降低，大脑区域的体积也减少。diceCT 后对组织进行尼氏染色和免疫荧光 Iba1 染色是成功的，但 RNA 提取不成功。因此，diceCT 显示出在 ZIKV 感染的小鼠发育中的大脑中检测 3D 定性和定量变化的有用性，并且在 diceCT 之后，还保留了对组织应用传统组织学和免疫荧光分析的能力。
