Arrangement of radial actin bundles in the growth cone of Aplysia bag cell neurons shows the immediate past history of filopodial behavior.

Filopodia that protrude forward from the lamellipodium, located at the leading edge of a neuronal growth cone, are needed to guide the extension of a nerve cell. At the core of each filopodium an actin bundle forms and grows into the lamellipodium. By using kymographs of time-lapse polarized light images we examined the relationship between the behavior of the filopodia, the actin bundles immediately proximal to the filopodia, and the shapes and composition of actin bundles in the whole lamellipodium. We find that the shapes of actin bundles, such as tilt, fork, and fused zones, originate at the leading edge and are surprisingly well preserved during retrograde transport of the actin cytoskeleton in the whole lamellipodium. The number of filaments that make up the radial actin bundles, as displayed by their birefringence retardation, also is preserved during retrograde flow over a distance of 4-8 microm from the leading edge into the lamellipodium. Thus, the disposition of the actin bundles in the lamellipodium frozen at any time point preserves and portrays a history of the past behavior of actin bundles proximal to the filopodia and the behavior of the filopodia themselves. These findings suggest that the arrangement of actin bundles in static image records, such as electron or fluorescence micrographs of fixed and stained specimens, can in fact reveal the sequence of the past history of filopodial behavior and the generation, density, fusion, etc. of the filaments in the actin bundles.

[1]  J. White,et al.  Cortical flow in animal cells. , 1988, Science.

[2]  Y. Wang,et al.  Exchange of actin subunits at the leading edge of living fibroblasts: possible role of treadmilling , 1985, The Journal of cell biology.

[3]  P. Forscher,et al.  An Aplysia cell adhesion molecule associated with site-directed actin filament assembly in neuronal growth cones. , 1996, Journal of cell science.

[4]  P. Bridgman,et al.  Nerve growth cone lamellipodia contain two populations of actin filaments that differ in organization and polarity , 1992, The Journal of cell biology.

[5]  N. K. Wessells,et al.  ULTRASTRUCTURE AND FUNCTION OF GROWTH CONES AND AXONS OF CULTURED NERVE CELLS , 1971, The Journal of cell biology.

[6]  S. J. Smith,et al.  Neuronal cytomechanics: the actin-based motility of growth cones. , 1988, Science.

[7]  C. S. Izzard,et al.  A precursor of the focal contact in cultured fibroblasts. , 1988, Cell motility and the cytoskeleton.

[8]  C. Cohan,et al.  Actin dynamics and organization during growth cone morphogenesis in Helisoma neurons. , 1997, Cell motility and the cytoskeleton.

[9]  P. Forscher,et al.  Myosin Drives Retrograde F-Actin Flow in Neuronal Growth Cones , 1996, Neuron.

[10]  T. Mitchison,et al.  Actin-Based Cell Motility and Cell Locomotion , 1996, Cell.

[11]  N. Hirokawa,et al.  Actin dynamics in growth cones , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[12]  L. Kaczmarek,et al.  Recruitment of Ca2+ channels by protein kinase C during rapid formation of putative neuropeptide release sites in isolated Aplysia neurons , 1992, Neuron.

[13]  E. Salmon,et al.  Quantifying single and bundled microtubules with the polarized light microscope. , 1995, The Biological bulletin.

[14]  P. Forscher,et al.  Cytoskeletal remodeling during growth cone-target interactions , 1993, The Journal of cell biology.

[15]  E D Salmon,et al.  Birefringence of single and bundled microtubules. , 1998, Biophysical journal.

[16]  R. Oldenbourg,et al.  New polarized light microscope with precision universal compensator , 1995, Journal of microscopy.

[17]  N. K. Wessells,et al.  Neuronal motility: the ultrastructure of veils and microspikes correlates with their motile activities. , 1983, Journal of cell science.

[18]  Mnh,et al.  Histologie du Système Nerveux de Lʼhomme et des Vertébrés , 1998 .

[19]  T. Mitchison,et al.  Actin dynamics in vivo. , 1997, Current opinion in cell biology.

[20]  M. Dailey,et al.  The organization of myosin and actin in rapid frozen nerve growth cones , 1989, The Journal of cell biology.

[21]  L. Kaczmarek,et al.  The morphology and coupling of Aplysia bag cells within the abdominal ganglion and in cell culture. , 1979, Journal of neurobiology.

[22]  P. J. Smith,et al.  Birefringence imaging directly reveals architectural dynamics of filamentous actin in living growth cones. , 1999, Molecular biology of the cell.

[23]  P. Forscher,et al.  Growth cone advance is inversely proportional to retrograde F-actin flow , 1995, Neuron.

[24]  A. Pestronk Histology of the Nervous System of Man and Vertebrates , 1997, Neurology.

[25]  R. Oldenbourg A new view on polarization microscopy , 1996, Nature.

[26]  P. Forscher,et al.  In vitro motility of immunoadsorbed brain myosin-V using a Limulus acrosomal process and optical tweezer-based assay. , 1995, Journal of cell science.

[27]  Julie A. Theriot,et al.  Actin microfilament dynamics in locomoting cells , 1991, Nature.

[28]  S. J. Smith,et al.  Actions of cytochalasins on the organization of actin filaments and microtubules in a neuronal growth cone , 1988, The Journal of cell biology.

[29]  P. Forscher,et al.  Novel form of growth cone motility involving site-directed actin filament assembly , 1992, Nature.