Differential sensitivity of cultured tanycytes and astrocytes to hydrogen peroxide toxicity

Tanycytes present in the mediobasal hypothalamus are able to support axonal regeneration and neuron survival. Pilot experiments of transplantation of these cells into various lesioned areas of the central nervous system (CNS) were thus performed to determine whether these cells could support the regeneration of the lesioned axons. These pilot experiments, however, demonstrated that the grafted tanycytes failed to survive in the lesioned sites. The present study was designed to determine which of the compounds released at the lesion would be toxic for tanycytes. Tanycyte cultures obtained from the median eminence of 10-day-old rats and astrocyte cultures obtained from the cortex of 10-day-old rats or E-14 embryos were incubated with two types of toxic molecules, including excitatory amino acids (EAA) and hydrogen peroxide (H2O2). The effect of these substances on cell death was estimated by measuring the lactate deshydrogenase (LDH) released and the surface occupied by immunostained glial structures after each treatment. The results indicated that the viability of both the tanycytes and the astrocytes was not affected by incubation for 24 h with 1 mM glutamate or 1 mM kainate. In contrast, increasing concentrations of H2O2 induced concentration-dependent cell death of tanycytes and immature astrocytes, without affecting the mature astrocytes. The use of antioxidant molecules such as catalase, tempol, or vitamin C effectively protected cultured tanycytes from H2O2 toxicity. These data indicate that (1) both mature astrocytes and tanycytes are resistant to EAA and (2) contrary to mature astrocytes, immature astrocytes and tanycytes are sensitive to the free radicals generated by H2O2. This suggest that oxidative stress is at least partly responsible for the death of tanycytes grafted into the lesioned CNS.