Τόμος 25 (2007) – Τεύχος 1 – Άρθρο 6 – Επιθεώρηση Κλινικής Φαρμακολογίας και Φαρμακοκινητικής-Ελληνική Έκδοση – Volume 25 (2007) – Issue 1– Article 6 – Epitheorese Klinikes Farmakologias και Farmakokinetikes-Greek Edition

Μελέτη της Επίδρασης Ανοξίας και Υπογλυκαιμίας στην Έκφραση Νευρωνικών Δεικτών σε Αμφιβληστροειδή και Ιππόκαμπο Επίμυος: Ανάπτυξη ex vivo Μοντέλου Ισχαιμίας

Effect of anoxia and hypoglycaemia on the expression of neuronal cell markers in the rat retina and hippo­campus: Development of a rat ex vivo ischemia model

Ελληνικά – Greek

Άρθρο

Article

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Neuronal ischemia leads to neuronal cells death due to glucose and oxygen deprivation. The goal of this study was the development of a rat ex vivo ischemia model for the deferential identification of anoxic and hypogly­caemic actions of ischemia on neuronal tissues which could subsequently used for the screening of possible neuroprotective agents.

Methods: The protocol used was a slight modification of one previously reported. In brief, the retina and 500μm thick hippocampal sections were preincubated for 30min in artificial cerebrospinal fluid pH 7.3 (ar­tiCSF) previously buffered with 95%Ο₂/5%CO₂. Tis­sues were subsequently incubated in artiCSF previ­ously buffered with 95%Ο₂/5%CO₂ (control and hypogly­caema) or 95%N₂/5%CO₂ (anoxia) in the pres­ence (control and anoxic conditions) or absence of glu­cose (hypoglycaemia: replacement with sucrose). The effect of anoxia and hypoglycaemia on neuronal cell markers’ expression was studied immunohistochemi­cally using antibodies against choline acetyl-trans­ferase (ChAT), γ-amino-butyric acid (GABΑ), neuronal nitric oxide synthase (bNOS) and neurofilament light (NF-L).

Results/Conclusions: In the retina, ex vivo hypoglycae­mic treatment led to the decrease of ChAT and GABA immunoreactivity in amacrine cells. This decrease was more profound under anoxic conditions. bNOS and NF-L retinal expression as well as GABA hippocampal expression did not seem to be affected by either treat­ment. As far as ChAT, GABA and bNOS immunoreactiv­ity are concerned our results were in agreement with previous studies of in vivo pressure-induced and in vitro chemical-induced retinal ischemia as well as transient cerebral ischemia. On the other hand, NF-L expression failed to correlate to the de­crease of NF-L mRNA levels observed after pressure-induced retinal ischemia/reperfusion. Although further studies are necessary for the assessment of the hypogly­caemia/hypoxia effects on the hippocampal neuronal marker expression this study led to the develop­ment of a conven­ient ex vivo retinal model for the screening of possible neuroprotective agents.