FREEZE-FRACTURING OF NERVE GROWTH CONES AND YOUNG FIBERS A Study of Developing Plasma Membr.pdf

FREEZE-FRACTURING OF NERVE GROWTH CONES AND YOUNG FIBERS A Study of Developing Plasma Membrane KARL H. PFENNINGER and RICHARD P. BUNGE From the Department of Anatomy, Washington University School of Medicine, St. Louis, Missouri 63 i 10. Dr. Pfenningers present address is Section of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510. ABSTRACT Neural and non-neural cellular processes have been studied in organotypic cultures of spinal cord and olfactory bulb by means of the freeze-fracturing technique. Identification of specific cellular elements in replicas has been achieved by comparison with thin-sectioned material in which differences in shape and contents are evident. Freeze-fracturing reveals that neural growth cones may be distin- guished from glial pseudopodia by the low number of intramembranous particles within their plasma membrane; the counts of particles within the growth cone membrane average 85/~tm 2 (for the inner leaflet) as opposed to hundreds per square micrometer in glial pseudopodia. Whereas the intramembranous particle number in glial pseudopodia is only slightly lower than in their perikaryal plasmalemma, the number of particles in outgrowing axons increases about eightfold from the periphery towards the perikaryon. Furthermore, with prolonged time of growth in culture, the particle density in the young nerve fibers increases by about the same factor. The same phenomenon, i.e. a low intramembranous particle level at earlier stages and an increase in numbers as the nerve fiber matures, is observed in fetal nerve tissue in vivo. These findings suggest that the plasmalemma of the outgrowing nerve, and especially of the growth cone, is immature and that maturation is accompanied by the insertion of intramembranous particles. Furthermore, these data indicate that the chemistry of the growth cone membrane is distinct from that of the neuron soma which may be significant for the mechanisms of gui