Expression of ischemia/reperfusion-inducible inflammatory markers and infarct volume are increased in regulator of calcineurin (Rcan)1 knockout mice after transient focal cerebral ischemia in mice. (A)
TNFα, IL-6 and Cox-2 mRNAs were amplified by TaqMan real time qRT-PCR from total RNA obtained from infarcted (I) and corresponding contralateral hemispheres (C) of wild-type (WT) (black columns) and Rcan1 knockout (KO) (gray columns) mice subjected to 90 minutes middle cerebral artery occlusion (MCAO) followed by 5 h reperfusion. Transcript amounts are normalized to TATA-binding protein (TBP) as an endogenous control, and are expressed relative to the level in contralateral samples from sham-operated animals after reperfusion for 5 h (sham). Real time qPCR was conducted in triplicate for each condition, and data are the means ± SD of four experiments. **P < 0.01, *P < 0.05 (ANOVA) versus contralateral sham samples. (B) Calcineurin (CN) enzyme activity against phosphopeptide RII measured in brain cortices from Rcan1 WT and Rcan1 KO mice. ns = non-significant (Student's t test). (C) Mice were subjected to 90 minutes MCAO followed by 48 h blood reperfusion, and infarct volumes were estimated by Cavalieri's principle from Nissl-stained serial coronal sections. (i) Representative stacks of six Nissl-stained sections, revealing a larger hypochromatic area of ischemic tissue in the infarcted neocortex of Rcan1 KO animals. (ii) Quantification of cerebral infarct volume in WT and Rcan1 KO mice. The percentage of tissue volume infarcted in the right hemisphere = (1 - (RN/L) ×100), where RN is spared tissue in right (infarcted) hemispheres and L is the left (contralateral) hemisphere; cerebral tissue volume is expressed in mm3. Data are means ± SEM. n = 7 per genotype; *P < 0.05.