A novel method for primary neuronal culture and characterization under different high temperature

Heatstroke is a big threat to human health; however, the characteristic of pathological changes of neurons during heatstroke development remains unclear. Here, using an in vitro model of primary cultured neurons from newborn Wistar rats, we investigated the effects of the different combinations of high temperature (37, 39, 41, 43, 45, and 47°C) and exposure time (45 min and 1 h) on the neurons. We found that, under the treatment of 45 min-heat, the neurons could resist high temperature up to 45°C, and under the treatment of 1 h-heat, the mortality of neurons increased as the temperature rises. After heating for 1 h, only a small minority of the neurons died under 41 and 43°C, which primarily occurred in the form of apoptosis. Up to 45°C for 1 h, most neurons occurred to necrosis. Meaningfully, some necrotic neurons expressed specific fried egg-like morphology. Our findings suggest that different high temperatures and exposure times were two key factors influencing the death of neurons. Under the high temperature (below 43°C for 1 h) similar to heatstroke, it just led a small percentage of neurons to apoptosis, and anti-apoptosis controls for preventing and treating heatstroke are promising.