Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

摘要

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

摘要

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

引言

The Centers for Disease Control and Injury Prevention estimate that approximately 2 million Americans sustain a traumatic brain injury (TBI) every year^1,2. In fact, TBI contributes to over 30% of all injury related deaths in the United States with healthcare costs nearing $80 billion annually and almost $4 million per person per year surviving a severe TBI^3,4,5. The impact of TBI is highlighted by the significant long-term neurocognitive and neuropsychiatric complications suffered by its survivors with the insidious o....

方案

All procedures were approved by the Northwestern University Institutional Animal Care and Use Committee. C57BL/6 mice were purchased from the Jackson Laboratory and group housed at a barrier facility at the Center for Comparative Medicine at Northwestern University (Chicago, IL). All animals were housed in 12/12 h light/dark cycle with free access to food and water.

1. Induce anesthesia

1. Anesthetize the mouse with ketamine (125 mg/kg) and xylazine (10 mg/kg) injected intraperitoneally.

2. Vital signs monitoring every 15 min

1. Monitor temperature, respiratory rate, and skin color. The m....

结果

The impactor mounts directly on the stereotaxic frame allowing for as much as 10 µm resolution for control of the point of impact, depth and penetration. The electromagnetic forces employed can impart impact velocities ranging 1.5–6 m/s. This allows for unparalleled precision and reproducibility over the entire range of clinically relevant TBI. Investigators can run pilot experiments changing the injury parameters such as impactor tip size, impact velocity, and impact depth to determine the parameters that best produce t.......

讨论

There are several steps that are critical for applying a reliable and consistent injury. First, the mouse must reach a deep plane of surgical anesthesia ensuring no movement during the performance of the craniectomy. While numerous anesthetic regimens may be used to induce general anesthesia in rodents, anesthetics that induce respiratory depression such as inhalational anesthetics may result in respiratory arrest when combined with a severe TBI. This protocol utilizes ketamine (125 mg/kg) and xylazine (10 mg/kg) injecte.......

材料

本文使用的材料清单

姓名	公司	目录编号	评论
AnaSed 注射液甲苯噻嗪无菌溶液	LLOYD， Inc.
丁丙诺啡 SR 实验室 0.5mg/mL	Zoopharm-Wildlife Pharmaceuticals USA	BSRLAB0.5-182012
高速旋转微电机 KiT0	Foredom Electric Company	K.1070
立体趋向性 CCI 的 Imapact one	Leica Biosystems Nussloch GmbH	39463920
Ketathesia 氯胺酮盐酸盐注射液 USP	Henry Schein， Inc	56344
小鼠特异性立体定位基	Leica Biosystems Nussloch GmbH	39462980
用于微钻的环钻	Fine Science Tools， Inc	18004-50