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Journal of Neuroscience, Vol 12, 2813-2818, Copyright © 1992 by Society for Neuroscience
A signal sequence mediates the retrograde transport of proteins from the axon periphery to the cell body and then into the nucleus
RT Ambron, R Schmied, CC Huang and M Smedman
Department of Anatomy and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
The presynaptic terminal and axon of neurons can undergo structural changes
in response to environmental signals. Since these changes require protein
synthesis in the cell body, the needs of the periphery must somehow be
communicated to the cell soma. To look for such a mechanism, we used
artificial protein constructs with properties expected of a signal that is
transported from the axon to the nucleus. One construct consisted of the
nuclear import signal peptide (sp) of the SV40 large T antigen, coupled to
human serum albumin (HSA) and rhodamine (r). When injected into the
axoplasm of Aplysia californica neurons in vitro, the rHSA-sp was
transported in the retrograde direction through the axon to the cell body
and then into the nucleus. Little, if any, moved in the anterograde
direction toward growth cones. The retrograde movement of injected rHSA-sp
was rapid (greater than 25 mm/d) and depended upon intact microtubules. The
sp portion of rHSA-sp provided access to both the retrograde transport
system and the nuclear import apparatus. Thus, rHSA was not transported at
all, but accumulated in organelles near the injection site. Also, rHSA-sp
containing an sp with a Lys to Thr substitution, which is known to reduce
nuclear import markedly, was transported only poorly. To look for
endogenous molecules that use this system, we affinity-purified a rabbit
polyclonal antibody to the signal sequence. The antibody recognized an 83
kDa polypeptide on Western blots of Aplysia nervous tissue. These data
indicate that Aplysia neurons contain the machinery to convey
macromolecules from the axon periphery to the nucleus.
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