Neutral red staining for the assessment of acute outcome in rat focal cerebral ischemia models.

Abstract

Neutral red staining was evaluated as an acute outcome assessment method in rat models of cerebral ischemia by comparison with histological infarction volume. Fischer 344 rats (n = 48) were used in three different models of middle cerebral artery (MCA) occlusion: proximal MCA occlusion (n = 16), distal MCA occlusion followed by ipsilateral common carotid artery (CCA) occlusion (distal MCA/CCA occlusion, n = 15), and MCA occlusion with an intravascularly introduced 4-0 nylon suture (intravascular MCA occlusion, n = 17). At 1 hour, 2 hours, and 4 hours after MCA occlusion, animals were injected with 2.5 ml of 4% neutral red solution via the femoral vein, and then sacrificed. Proximal MCA occlusion caused a neutral red defect volume in the cortex which correlated well with histological infarction volume at 4 hours (r = 0.88, p < 0.05), and in the caudate which correlated well with infarction volume at 4 hours (r = 0.94, p < 0.01). Distal MCA/CCA occlusion caused a neutral red defect volume in the cortex lager than the histological infarction volume (4 hrs: 88.6 +/- 11.8 vs. 74.3 +/- 17.4 mm3, p < 0.05) but closely correlated with the infarction volume at 4 hours (r = 0.81, p < 0.05). Intravascular MCA occlusion caused a neutral red defect volume in only two of 17 animals after 1-4 hours, which correlated well with the absence of histological evidence of infarction. Neutral red staining is a simple method for assessing the acute outcome of focal cerebral ischemia as early as 4 hours after the onset, in an appropriate model of cerebral ischemia.