Association of microtubule-associated protein (MAP1B) with growing axons in cultured hippocampal neurons.
Microtubule-associated protein 1B (MAP1B) is a major constituent of the neuronal cytoskeleton early in development. This protein is present in embryonic brain and is composed of two isoforms that are the result of differential phosphorylation. We examined the distribution of MAP1B during the differentiation of cultured hippocampal neurons and compared it to that of MAP2 and tubulin. We demonstrated by immunofluorescent doublestaining that MAP1B and MAP2 are colocalized in cell bodies and the minor processes of hippocampal neurons during the early stages of development, before the establishment of neuronal polarity. Later, when neurons acquire axonal and dendritic characteristics, MAP1B is sorted into growing axons, including the growth cone, whereas MAP2 is restricted to dendrites and cell bodies. Unlike tubulin, the localization of MAP1B in growing axons is not uniform. Rather, the protein is found concentrated in the distal portion. During later stages of development, the neurons extend a network of fasciculating axonal and dendritic neurites in which the segregation of MAP1B and MAP2 is maintained. However, the staining of MAP1B in mature neuronal cultures decreases in a pattern that resembles the decline of this protein during brain development. These results support the association of MAP1B with growing axons and its correct developmental regulation in the hippocampal culture system.
Fischer
I
I
Department of Biochemistry, E. K. Shriver Center, Waltham, Massachusetts 02254, USA; Department of Neurology (Neuroscience), Harvard Medical School, Boston, Massachusetts 02115, USA.
Romano-Clarke
G
G