Selective expression of JNK isoforms and stress-specific JNK activity in different neural cell lines

Abstract

The function of c-Jun N-terminal kinases (JNKs) in the nervous system is poorly understood and the majority of the data has been gained in neuronal and non-neuronal cell lines. Thus, it is not clear to which extent the expression pattern and the degree of activation of the three JNK isoforms in different cell lines are representative for their activation in the adult brain. In the present study, the expression of JNK isoforms and the activity of JNK1 were determined following UV irradiation and exposure to H 2O 2 and TNFα in three neural cell lines, rat PC12, murine Neuro2A and human SHSY5Y. These cell lines differ in their expression of JNK isoforms: PC12 cells express JNK1 and JNK2, whereas Neuro2A and SHSY5Y cells displays the expression of JNK1, JNK2 and JNK3. JNK3 was not inducible following stress and differentiation in PC12 cells. The stimulation paradigms evoked different degree of cell death: UV irradiation resulted in death of around 50% in all three cell lines; exposure to 200 μM H 2O 2 for 6 h resulted in the death of 43% Neuro2A cells and 31% PC12 cells, SHSY5Y cells are less sensitive to H 2O 2 since only 5 mM H 2O 2 killed 59% of SHSY5Y cells after 6 h. Exposure to 50 ng/ml TNFα did not induce cell death in SHSY5Y, Neuro2A and naive PC12 cells. Although differentiated PC12 cells exhibit a similar activation of JNK1 compared to naive PC12 cells after exposure to TNFα, 42% of differentiated PC12 cells died after 24 h. H 2O 2 that evoked only a moderate JNK1 activity in Neuro2A and PC12 cells induced only a moderate cell death. In contrast, SHSY5Y cells exhibit a much stronger JNK1 activation accompanied with a higher degree in cell death after exposure to H 2O 2. JNK1 activity induced by UV irradiation, however, could not be correlated with the extend of cell death. These data clearly demonstrate that expression and activation of JNK depends on the neuronal cell type and the applied stress paradigms, and that JNK activity is not simply linked to cell death.