Use of confocal microscopy in the study of ischemia-induced hippocampal neuronal damage

The present study was undertaken to reveal by means of confocal laser microscopy the cytoarchitecture of hippocampal CA3 neurons in Mongolian gerbils before and after cerebral ischemia of different duration. The common carotid arteries of gerbils were occluded for 5, 10, or 15 min. On the 4th, 14th and 28th day after reperfusion, neuronal damage was examined by laser scanning confocal microscopy in the CA3 region of hippocampus (30 ?m slices). Slices were stained with fluorescent Nissl staining and fluorescent membrane tracer DiI. Increased duration of cerebral ischemia resulted in a progressive loss of hippocampal CA3 neurons. Four days after the ischemic insult, neuronal damage in the hippocampal CA3 region was mild but visible. On the 28th day after reperfusion, neuronal damage in the observed brain structure was most severe. These results demonstrate the temporal profile of neuronal damage after an ischemic insult as observed using confocal microscopy. Eksperimentalni model za izucavanje ishemije bili su dzerbili - pustinjski misevi, koji su bili izlozeni 5, 10 ili 15-minutnoj ishemiji. Cetvrtog, 14 i 28 dana posle reperfuzije stepen ostecenja neurona u CA3 regionu hipokampusa je procenjivan primenom konfokalne mikroskopije (dvostruko bojenje) na 30?m presecima. Nasi rezultati pokazuju da je stepen ostecenja neurona zavistan od duzine ishemije i povecava se vremenom posle reperfuzije. Najveci stepen ostecenja neurona CA3 regiona hipokampusa bio je 28. dana posle 15-minutne ishemije.