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Vol. 10, Issue 9, 2919-2931, September 1999
*San Raffaele Scientific Institute, Consiglio Nazionale
delle Richerche Center for Cellular and Molecular Pharmacology
and B. Ceccarelli Center for Neurobiology, University of Milan, Milan,
Italy; We have investigated the process leading to differentiation of PC12
cells. This process is known to include extension of neurites and
changes in the expression of subsets of proteins involved in
cytoskeletal rearrangements or in neurosecretion. To this aim, we have
studied a PC12 clone (trk-PC12) stably transfected with the nerve
growth factor receptor TrkA. These cells are able to undergo both
spontaneous and neurotrophin-induced morphological differentiation. However, both undifferentiated and nerve growth factor-differentiated trk-PC12 cells appear to be completely defective in the expression of proteins of the secretory apparatus, including proteins of synaptic vesicles and large dense-core granules,
neurotransmitter transporters, and neurotransmitter-synthesizing
enzymes. These results indicate that neurite extension can occur
independently of the presence of the neurosecretory machinery,
including the proteins that constitute the fusion machine, suggesting
the existence of differential activation pathways for the two processes
during neuronal differentiation. These findings have been confirmed in independent clones obtained from PC12-27, a previously characterized PC12 variant clone globally incompetent for regulated secretion. In
contrast, the integrity of the Rab cycle appears to be necessary for
neurite extension, because antisense oligonucleotides against the
neurospecific isoform of Rab-guanosine diphosphate-dissociation inhibitor significantly interfere with process formation.
Department of Neuroscience, University of Rome Tor
Vergata, Roma, Italy; and Consiglio Nazionale delle
Richerche Institute of Genetics, Biochemistry and Evolution, Pavia,
Italy
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