Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope

作者全名："Zhong, Jianjun; Gunner, Georgia; Henninger, Nils; Schafer, Dorothy P.; Bosco, Daryl A."

作者地址："[Zhong, Jianjun; Henninger, Nils; Bosco, Daryl A.] Univ Massachusetts, Chan Med Sch, Dept Neurol, Worcester, MA 01655 USA; [Zhong, Jianjun] Chongqing Med Univ, Affiliated Hosp 1, Dept Neurosurg, Chongqing, Peoples R China; [Gunner, Georgia; Schafer, Dorothy P.] Univ Massachusetts, Chan Med Sch, Dept Neurobiol, Worcester, MA USA"

通信作者："Bosco, DA (通讯作者)，Univ Massachusetts, Chan Med Sch, Dept Neurol, Worcester, MA 01655 USA."

来源：JOVE-JOURNAL OF VISUALIZED EXPERIMENTS

ESI学科分类：Multidisciplinary

WOS号：WOS:000994475300010

JCR分区：Q3

影响因子：1.2

年份：2023

卷号：　

期号：194

开始页：　

结束页：　

文献类型：Article

关键词：　

摘要："The goal of this protocol is to demonstrate how to longitudinally visualize the expression and localization of a protein of interest within specific cell types of an animal's brain, upon exposure to exogenous stimuli. Here, the administration of a closed-skull traumatic brain injury (TBI) and simultaneous implantation of a cranial window for subsequent longitudinal intravital imaging in mice is shown. Mice are intracranially injected with an adeno-associated virus (AAV) expressing enhanced green fluorescent protein (EGFP) under a neuronal specific promoter. After 2 to 4 weeks, the mice are subjected to a repetitive TBI using a weight drop device over the AAV injection location. Within the same surgical session, the mice are implanted with a metal headpost and then a glass cranial window over the TBI impacting site. The expression and cellular localization of EGFP is examined using a two-photon microscope in the same brain region exposed to trauma over the course of months."

基金机构：Department of Defense; [W81XWH202071/PRARP]

基金资助正文："We thank Dr. Miguel Sena-Esteves at the University of Massachusetts Chan Medical School for gifting the AAV (PHP.eB) -Syn1-EGFP virus, and Debra Cameron at the University of Massachusetts Chan Medical School for drawing the mice skull sketch. We also thank current and past members of the Bosco, Schafer and Henninger labs for their suggestions and support. This work was funded by the Department of Defense (W81XWH202071/PRARP) to DAB, DS, and NH."