A Mouse Controlled Cortical Impact Model of Traumatic Brain Injury for Studying Blood–Brain Barrier Dysfunctions
Traumatic brain injury (TBI) is one of the leading causes of death and disability worldwide. It is a silently growing epidemic with multifaceted pathogenesis, and current standards of treatments aim to target only the symptoms of the primary injury, while there is a tremendous need to explore interventions that can halt the progression of the secondary injuries. The use of a reliable animal model to study and understand the various aspects the pathobiology of TBI is extremely important in therapeutic drug development against TBI-associated complications. The controlled cortical impact (CCI) model of TBI described here, uses a mechanical impactor to inflict a mechanical injury into the mouse brain. This method is a reliable and reproducible approach to inflict mild, moderate or severe injuries to the animal for studying TBI-associated blood–brain barrier (BBB) dysfunctions, neuronal injuries, brain edema, neurobehavioral changes, etc. The present method describes how the CCI model could be utilized for determining the BBB dysfunction and hyperpermeability associated with TBI. Blood–brain barrier disruption is a hallmark feature of the secondary injury that occur following TBI, frequently associated with leakage of fluid and proteins into the extravascular space leading to vasogenic edema and elevation of intracranial pressure. The method described here focuses on the development of a CCI-based mouse model of TBI followed by the evaluation of BBB integrity and permeability by intravital microscopy as well as Evans Blue extravasation assay.
Key wordsBlood–brain barrier Controlled cortical impact Traumatic brain injury Intravital microscopy Evans Blue Edema Intracranial pressure Central nervous system Hyperpermeability Endothelial permeability
The work presented in this chapter was supported by Scott & White Academic Operations Funds to Dr. Tharakan. The authors would like to apologize to investigators whose works are not cited in this methodological report due to space limitations and the personal perspective with which this chapter has been prepared.
- 7.Thurman DJ, Branche CM, Sniezek JE (1998) The epidemiology of sports-related traumatic brain injuries in the United States: recent developments. J Head Trauma Rehabil 3:1–8Google Scholar
- 8.Scott BN, Roberts DJ, Robertson HL, Kramer AH, Laupland KB, Ousman SS, Kubes P, Zygun DA (2013) Incidence, prevalence, and occurrence rate of infection among adults hospitalized after traumatic brain injury: study protocol for a systematic review and meta-analysis. Syst Rev 2:68CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Kasper C, Yvette C (2015) Traumatic brain injury. In: Kasper C, Conley Y (eds) Annual review of nursing research, vol 33. Springer, New YorkGoogle Scholar
- 17.Nidavani RB, Mahalakshmi AM, Shalawadi M (2014) Vascular permeability and Evans blue dye: a physiological and pharmacological approach. J Appl Pharm Sci 4:106–113Google Scholar
- 19.Khan M, Im YB, Shunmugavel A, Gilg AG, Dhindsa RK, Singh AK, Singh IJ (2009) Administration of S-nitroglutathione after traumatic brain injury protects the neurovascular unit and reduces secondary injury in a rat model of controlled cortical impact. J Neuroinflammation 6:32CrossRefPubMedPubMedCentralGoogle Scholar
- 20.Masedunskas A, Porat-Shliom N, Tora M, Milberg O, Weigert R (2013) Intravital microscopy for imaging subcellular structures in live mice expressing fluorescent proteins. J Vis Exp (79)Google Scholar
- 21.Marques PE, Oliveira AG, Amaral SS, Nunes-Silva A, Almeida AFS (2012) Intravital microscopy: taking a close look inside the living organisms. Afr J Microbiol Res 6:1603–1614Google Scholar
- 23.Radu M, Chernoff J (2013) An in vivo assay to test blood vessel permeability. J Vis Exp (73):e50062Google Scholar