Cytochrome oxidase is decreased in piglet hippocampus following hypoxia-ischemia

We assessed cytochrome oxidase (CytOx) staining in sham-operated control piglets and in piglets subjected to 30 minutes of cerebral hypoxia-ischemia (H-I) plus 4 hours of reperfusion (REP). The 1-day-old piglets were sedated, anesthetized, and ventilated. Cerebral blood flows (CBFs) were quantitated using microspheres. H-I was induced by a combination of phlebotomy and cervical tourniquet; the brain was reperfused for four hours after 30 minutes of H-I. CBF was reduced during ischemia in experimental animals from 42 + 13 to 12 + 5 ml/min/100g. CytOx staining of hippocampal sections from 3 control and 3 experimental animals was compared. The staining of the stratum pyramidale neurons of the same portion of the CA1 sector in a single high power field was assessed in a blinded fashion in 4 corresponding sections from each animal, and graded from 0 = no staining to 3 = heavy staining. The results were compared using one-way analysis of variance. Cells with grade 3 staining were significantly more numerous in controls compared to H-I/REP animals (p = 0.03). There were significantly more cells with no CytOx staining in the experimental animals (p = 0.01). These findings suggest that CytOx staining in newborn piglet CA1 is a reliable method of assessing cell dysfunction after H-I.