Abstract
Intracerebral infusion of kainic acid (KA) by a microdialysis probe induces a focal swelling in the brain-perfused area which promotes inflammation (Compan et al., 2012; Oprica et al., 2003). The microdialysis technique allows the local in vivo perfusion of KA and the simultaneous collection of inflammatory mediators, and other neuroactive substances, released in the injured brain. This protocol also allows the perfusion of different solutions in each cerebral hemisphere at the same time. By perfusing KA in isotonic solution of Krebs-Ringer Bicarbonate (KRB) (280-290 mOsm) in one hippocampus and KA in hypertonic KRB solution (1,400-1,500 mOsm) in the contralateral side, we can evaluate in vivo the efficiency of hypertonic solutions in preventing inflammation induced by swelling after KA infusion. Once the inflammatory response has been induced, it is possible to infuse through the microdialysis probe a biotinylated specific inhibitor of caspase-1 allowing the detection of the brain regions and cells involved in IL-1 production in response to the injury (Oprica et al., 2003).
Keywords: ELISA, In situ caspase-1 detection, Brain injury, Inflammasome, Microdyalisis
Materials and Reagents
Equipment
Experimental set-up
Procedure
Recipes
Acknowledgments
This protocol has been adapted from the previously published paper: Compan et al. (2012). This work was supported by grants from PN I+D+I 2008-2011-Instituto Salud Carlos III-FEDER (EMER07/049 and PI09/0120) and Fundación Séneca (11922/PI/09). The authors declare no conflict of interests.
References
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