Abstract
To study the ionic, metabolic, and morphologic derangements that occur following brain injury we utilized a retina in vitro model of hypoxia. Retinas were dissected into oxygenated (95% O2, 5% CO2) Ames medium, a physiologic solution resembling cerebrospinal fluid, and randomly assigned to either experimental hypoxic conditions (95% N2, 5% CO2) or control conditions. All retinas were incubated and maintained at 37 degrees C. Changes in extracellular K+ and lactate concentration, intracellular incorporation of 45Ca and 14C-leucine, uptake of glucose using [14C]-2-deoxy-D-glucose (2DG), and cell size were determined at 10, 20, 30, and 60 minute time intervals. The results show that compared to control retinas hypoxia produced: (1) an early increase in extracellular concentration of K+ and lactate, (2) a delayed increase in the intracellular incorporation of 45Ca, (3) an early onset of cellular swelling, and (4) a decrease in the intracellular incorporation of 14C-leucine, and (5) increased glucose utilization. All of the results were statistically significant (p < 0.05) and exhibited a dose response relationship with the exception of intracellular incorporation of 45Ca which did not become significantly different until 30 minutes post-hypoxia. A 16% increase in cell size was noted after 10 minutes of hypoxia. Increased hypoxic cell size persisted for 30 minutes but after 60 minutes the control retinas appeared enlarged as well. Our results suggest that ionic, metabolic, and morphologic derangements can be demonstrated utilizing an in vitro model of hypoxia which are similar to those seen following in vivo traumatic brain injury. With use of this model the mechanisms behind these ionic-metabolic relationships can be addressed at the molecular level.
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