Temperature Modulation of the Effects of Repetitive Anoxia on Potassium Homeostasis in the Brain of Drosophila melanogaster

Abstract

Oxygen can be limited at the environmental (e.g. flood-prone burrows) or cellular (e.g. stroke, heart attack) levels. O2 deprivation in nervous tissue depolarizes cell membranes, incrementing extracellular potassium concentration ([K+]o). Consequently, [K+]o can be used to assess neural failure during anoxia. The effect of temperature on the maintenance of brain [K+]o homeostasis in male and female Drosophila melanogaster (W1118) was assessed during repeated anoxic comas induced by N2 gas. Brain [K+]o was continuously monitored using K+-sensitive microelectrodes while body temperature was gradually increased/decreased using a Peltier plate. Once the desired temperature was reached (16°C/17°C, 23°C or 29°C/30°C), it was maintained for the rest of the experiment and the fly was subjected to repeated anoxic bouts. Repetitive anoxia resulted in a loss of the ability to maintain [K+]o baseline at ~10 mM. In both sexes, the total [K+]o baseline variation (D[K+]o) was augmented at 30°C (D[K+]o male = 119.2 ± 21.9 mM; D[K+]o female = 51.2 ± 8.1 mM), whereas 16°C stabilized [K+]o baseline for the duration of the experiment (D[K+]o male = 17.5 ± 4.1 mM; D[K+]o female = 16.9 ± 6.8 mM). Additionally, D[K+]o in males was significantly greater (114.3 ± 10.5 mM ) than in females (36.1 ± 10.5 mM) at 23°C. Under reduced dehydration, experiments performed only in males showed the same trends although the D[K+]o values where considerably reduced at 17°C (D[K+]o male = -1.0 ± 1.3 mM) and 23°C (D[K+]o male = 17.3 ± 1.5 mM) and increased at 29°C (D[K+]o male = 332.7 ± 83.0 mM). It was concluded that 1) N2-delivery patterns consisting of long anoxia, short normoxia and high cycle frequency increased disruption of brain [K+]o baseline maintenance, 2) males were more susceptible to repeated anoxia than females at room temperature, and 3) hypothermia had a protective effect on brain K+ homeostasis during repetitive anoxia. Male flies are suggested as a useful model for examining deleterious consequences of O2 reperfusion with extensive application on therapeutical treatment of stroke or heart attack.