Longitudinal intravital microscopy of the mouse kidney: inflammatory responses to abdominal imaging windows.

Michelle M Martinez Irizarry, Julia R Walsh, Malgorzata M Kamocka, Hyowon Lee, Kenneth W Dunn
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Abstract

Intravital microscopy enables direct observation of cell biology and physiology at subcellular resolution in real time in living animals. Implanted windows extend the scope of intravital microscopy to processes extending for weeks or even months, such as disease progression or tumor development. However, a question that must be addressed in such studies is whether the imaging window, like any foreign body, triggers an inflammatory response, and if that response alters the biological process under investigation. To directly evaluate this question, we conducted large-scale intravital microscopy of the kidney of LysM-EGFP mice over time after implantation of abdominal imaging windows. These studies demonstrate that windows stimulated a variety of changes consistent with a foreign body response. Within a few days of implantation, leukocytes were recruited to the window and the region between the window and kidney where, over the next 16 days, they increased in number in an expanding volume that developed a new vascular network. These changes were accompanied by a dramatic increase in glomerular albumin permeability within 2 - 5 days of implantation. Similar results were obtained from mice implanted with windows coated with PLL-g-PEG, but not from immune-deficient mice. These studies demonstrate the importance of evaluating whether implanted windows induce an inflammatory response, and whether that response impacts the processes under evaluation in longitudinal intravital microscopy studies.

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小鼠肾脏的纵向体视显微镜检查:腹部成像窗口的炎症反应。
体视显微镜可在活体动物体内以亚细胞分辨率实时直接观察细胞生物学和生理学。植入式成像窗将体视显微镜的观察范围扩展到了延续数周甚至数月的过程,如疾病进展或肿瘤发展。然而,此类研究必须解决的一个问题是,成像窗口是否会像任何异物一样引发炎症反应,以及这种反应是否会改变所研究的生物过程。为了直接评估这个问题,我们对植入腹部成像窗后一段时间内的 LysM-EGFP 小鼠肾脏进行了大规模的体视显微镜观察。这些研究表明,腹腔成像窗刺激了与异物反应一致的各种变化。在植入后的几天内,白细胞被招募到窗口和窗口与肾脏之间的区域,在接下来的 16 天内,白细胞数量不断增加,体积不断扩大,形成了新的血管网络。伴随着这些变化,肾小球白蛋白通透性在植入后 2-5 天内急剧增加。植入涂有 PLL-g-PEG 的窗口的小鼠也获得了类似的结果,但免疫缺陷小鼠却没有。这些研究表明,评估植入的窗口是否会诱发炎症反应以及这种反应是否会影响纵向观察显微镜研究中的评估过程非常重要。
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