Raf-1 and B-Raf proteins have similar regional distributions but differential subcellular localization in adult rat brain

The Raf kinases play an important and specific role in the activation of ex tracellular signal-regulated kinases (ERK) cascade. Beside its role in the control of proliferation and differentiation, the ERK cascade has also been implicated in neuron-specific functions. In order to gain clues on the fun ction of Raf kinases in the adult central nervous system (CNS), we performe d a comparative analysis of the distribution and subcellular localization o f the different Raf kinases in rat brain with antibodies specific for the d ifferent Raf kinases. We show that B-Raf and Raf-l proteins are present in most brain areas, whereas A-Raf is not detected. Interestingly, the two Raf proteins have an approximately similar pattern of distribution with a rost ro-caudal decreasing gradient of expression. These two kinases are colocali zed in neurons but they are differentially located in subcellular compartme nts. Raf-l is localized mainly in the cytosolic fraction around the nucleus , whereas B-Raf is widely distributed in the cell bodies and in the neuriti c processes. In addition, we demonstrated that numerous B-Raf isoforms are present in the brain. These isoforms have a differential pattern of distrib ution, some of them being ubiquitously expressed whereas others are localiz ed to specific brain areas. These isoforms also have a clear differential s ubcellular localization, specially in Triton-insoluble fractions, but also in synaptosomal, membrane and cytosolic compartments. Altogether these resu lts suggest that each Raf protein could have a distinct signalling regulato ry function in the brain with regard to its subcellular localization.