The topography and subcellular distribution ofmitogen-activated protein kinase kinase 1 (MEK1) inadult rat brain and differentiating PC12 cells

Abstract

Mitogen-activated protein kinase signal transduction pathway involved in the regulation of proliferation and differentiation of various mammalian cells consists of a sequential activation of three protein kinases, Raf, mitogen-activated protein kinase kinase, and mitogen-activated protein kinase. These kinases are highly expressed in brain and play an important role in neuronal signalling. In this study, to further characterize mitogen-activated protein kinase signalling pathway in brain, we have elucidated the topography and subcellular distribution of mitogen-activated protein kinase kinase1 in adult rat brain and differentiating PC12 cells. Our immunohistochemical data indicate that mitogen-activated protein kinase kinase1 is widely distributed throughout the brain and expressed prominently in cortex, hippocampus, brainstem, hypothalamus and cerebellum. In these areas of brain mitogen-activated protein kinase kinase1 is exclusively neuronal in origin and is localized within perikarya and dendrites. Confocal microscopy data has determined that a portion of mitogen-activated protein kinase kinase1 in rat brain is co-localized with microtubules. This co-localization was observed only within neuritic shaft and cilia of ventricular ependymal cells. In nerve growth factor-induced differentiating PC12 cells, mitogen-activated protein kinase kinase1 displays co-localization with microtubules within proximal regions of neuritic shafts and their junctions with the cell somas. From bovine brain extract, mitogen-activated protein kinase kinase1 co-purifies with microtubules. In vitro kinase assay detected mitogen-activated protein kinase kinase1 activity within purified microtubules. These observations indicate that mitogen-activated protein kinase kinase1 is associated with microtubules within some specialized compartments of the brain and microtubule-associated mitogen-activated protein kinase kinase1 is catalytically active.