Department of Neurology and Neurological Sciences, Stanford University School of Medicine
This article is a part ofJoVE General. If you think this article would be useful for your research, please recommend JoVE to your institution's librarian.Recommend JoVE to Your Librarian
Current Access Through Your IP Address
Current Access Through Your Registered Email Address
Taniguchi, H., Andreasson, K. The Hypoxic Ischemic Encephalopathy Model of Perinatal Ischemia. J. Vis. Exp. (21), e955, doi:10.3791/955 (2008).
Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe and permanent neuropsychological sequelae, including mental retardation, cerebral palsy, and epilepsy. The outcomes of HIE are devastating and permanent, making it critical to identify and develop therapeutic strategies to reduce brain injury in newborns with HIE. To that end, the neonatal rat model for hypoxic-ischemic brain injury has been developed to model this human condition. The HIE model was first validated by Vannucci et al 1 and has since been extensively used to identify mechanisms of brain injury resulting from perinatal hypoxia-ischemia 2 and to test potential therapeutic interventions 3,4. The HIE model is a two step process and involves the ligation of the left common carotid artery followed by exposure to a hypoxic environment. Cerebral blood flow (CBF) in the hemisphere ipsilateral to the ligated carotid artery does not decrease because of the collateral blood flow via the circle of Willis; however with lower oxygen tension, the CBF in the ipsilateral hemisphere decreases significantly and results in unilateral ischemic injury. The use of 2,3,5-triphenyltetrazolium chloride (TTC) to stain and identify ischemic brain tissue was originally developed for adult models of rodent cerebral ischemia 5, and is used to evaluate the extent of cerebral infarctin at early time points up to 72 hours after the ischemic event 6. In this video, we demonstrate the hypoxic-ischemic injury model in postnatal rat brain and the evaluation of the infarct size using TTC staining.
This protocol was approved by the Institutional Animal Care and Use Committee at Stanford University and abides by the National Institutes of Health guidelines for the use of experimental animals.
Neonatal Rat HIE model
Figure 1. Representative HIE coronal levels 1-4 stained with TTC.
TTC stained coronal sections demonstrate area of infarct (in white).
The rodent postnatal HIE model is an established model that recapitulates cerebral hypoxia occurring in the peri-natal period in newborns. Extensive histological and immunocytochemical characterization studies have demonstrated that the 7 day old rodent has the analogous brain maturity to model a third trimester human fetus 7. The rodent HIE model has been very informative for understanding mechanisms of brain injury from peri-natal hypoxia 2, where interventions such as hypothermia have been validated 3. This model was initially perfected in rats and more recently adapted to mice 8.
There are specific technical details that warrant mention:
Funded by American Heart Association and March of Dimes.
|Sprague-Dawley Rat Pups||Animal||Charles River Laboratories|
|Isoflurane||Surgery||Baxter Internationl Inc.|
|8% Oxygen/ 92% Nitrogen Gas||Surgery||Airgas|
|2,3,5-triphenyl tetrazolium chloride||Reagent||Sigma-Aldrich||T8877|
|Phosphate buffered saline (PBS) pH 7.4||Reagent||GIBCO, by Life Technologies|
1. J. E. Rice, 3rd, R. C. Vannucci, and J. B. Brierley, Annals of neurology 9 (2), 131 (1981).
2. R. C. Vannucci and S. J. Vannucci, Dev Neurosci 27 (2-4), 81 (2005).
3. E. Bona, H. Hagberg, E. M. Loberg et al., Pediatric research 43 (6), 738 (1998).
4. K. Mishima, T. Ikeda, T. Yoshikawa et al., Behavioural brain research 151 (1-2), 209 (2004).
5. W. H. Trescher, S. Ishiwa, and M. V. Johnston, Brain & development 19 (5), 326 (1997);
6. R. S. Young, T. P. Olenginski, S. K. Yagel et al., Stroke; a journal of cerebral circulation 14 (6), 929 (1983).
7. J. B. Bederson, L. H. Pitts, S. M. Germano et al., Stroke; a journal of cerebral circulation 17 (6), 1304 (1986).
8. J. Cai, Z. Kang, W. W. Liu et al., Neuroscience letters 441 (2), 167 (2008).
9. J. Grow and J. D. Barks, Clinics in perinatology 29 (4), 585 (2002).
10. U. Aden, L. Halldner, H. Lagercrantz et al., Stroke; a journal of cerebral circulation 34 (3), 739 (2003).