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The development of a long, coiled, optic nerve in the stalk-eyed fly Cyrtodiopsis whitei


In the stalk-eyed fly Cyrtodiopsis whitei (Diopsidae; Diptera), the relatively long optic nerve develops within the tight lumen of a very short eyestalk. Axonal growth is generally considered in terms of path finding, selective fasciculation, and towing. Physical forces that are necessary for axon lengthening are generated either by the growth cone or by the growth of surrounding tissues. Therefore, it is surprising to encounter a loosely coiled nerve apparently lacking any attachments that could allow for pull, or towing, of the nerve. In this study, we used histological sections and whole-mount preparations to confirm that the optic nerve of the stalk-eyed fly indeed elongates without the external application of tension to the nerve. Secondly, we examined the distribution of cytoskeletal elements and selected proteins that may be involved in axon extension. Staining against the vesicle fusion proteins SNAP-24 and SNAP-25 consistently results in stronger staining in the rapidly extending optic nerve than in a control nerve, suggesting a possible role of these proteins in the extension process. On a gross morphological level, SNAP-24/25 as well as the cytoskeletal elements actin and tubulin are uniformly distributed throughout the lengths of the growing nerve, suggesting that nerve elongation is distributed rather than localized. Finally, we identified glia as a possible source for tension within the nerve bundle. Glia proliferate rapidly in the optic nerve but not in the control nerve. Much work continues to focus on the growth of axons in culture, but this study is one of the few that considers the dynamics of nerve bundle extension as a whole.

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  1. Auld V (1999) Glia as mediators of growth cone guidance: studies from insect nervous systems. Cell Mol Life Sci 55:1377–1385

  2. Bray D (1970) Surface movements during growth of single explanted neurons. Proc Natl Acad Sci U S A 65:905–910

  3. Bray D (1984) Axonal growth in response to experimentally applied mechanical tension. Dev Biol 102:379–389

  4. Buschbeck EK, Hoy RR (1998) Visual system of the stalk-eyed fly, Cyrtodiopsis quinqueguttata (Diopsidae, Diptera): an anatomical investigation of unusual eyes. J Neurobiol 37:449–468

  5. Buschbeck EK, Roosevelt JL, Hoy RR (2001) Eye stalks or no eye stalks: a structural comparison of pupal development in the stalk-eyed fly Cyrtodiopsis and in Drosophila. J Comp Neurol 433:486–498

  6. Craig AM, Wyborski RJ, Banker G (1995) Preferential addition of newly synthesized membrane-protein at axonal growth cones. Nature 375:592–594

  7. Dai J, Sheetz MP, Wan X, Morris CE (1998) Membrane tension in swelling and shrinking molluscan neurons. J Neurosci 18:6681–6692

  8. Dehmelt L, Halpain S (2004) Actin and microtubules in neurite initiation: are MAPs the missing link? J Neurobiol 58:18–33

  9. Dennerll TJ, Lamoureux P, Buxbaum RE, Heidemann SR (1989) The cytomechanics of axonal elongation and retraction. J Cell Biol 109:3073–3083

  10. Dent EW, Kalil K (2001) Axon branching requires interactions between dynamic microtubules and actin filaments. J Neurosci 21:9757–9769

  11. Ferreira A, Niclas J, Vale RD, Banker G, Kosik KS (1992) Suppression of kinesin expression in cultured hippocampal neurons using antisense oligonucleotides. J Cell Biol 117:595–606

  12. Futerman AH, Banker GA (1996) The economics of neurite outgrowth–the addition of new membrane to growing axons. Trends Neurosci 19:144–149

  13. Gallo G (1998) Involvement of microtubules in the regulation of neuronal growth cone morphologic remodeling. J Neurobiol 35:121–140

  14. Gordon-Weeks PR (2004) Microtubules and growth cone function. J Neurobiol 58:70–83

  15. Griffin JW, Price DL, Drachman DB, Morris J (1981) Incorporation of axonally transported glycoproteins into axolemma during nerve regeneration. J Cell Biol 88:205–214

  16. Grimaldi D, Fenster G (1989) Evolution of extreme sexual dimorphism: structural and behavioral convergence among broad-headed Drosophilidae (Diptera). Am Mus Novit 2939:1–25

  17. Heidemann SR (1996) Cytoplasmic mechanisms of axonal and dendritic growth in neurons. Int Rev Cyt 165:235–296

  18. Heidemann SR, Kaech S, Buxbaum RE, Matus A (1999) Direct observations of the mechanical behaviors of the cytoskeleton in living fibroblasts. J Cell Biol 145:109–122

  19. Herring TL, Cohan CS, Welnhofer EA, Mills LR, Morris CE (1999) F-actin at newly invaginated membrane in neurons: implications for surface area regulation. J Membr Biol 171:151–169

  20. Lamoureux P, Buxbaum RE, Heidemann SR (1989) Direct evidence that growth cones pull. Nature 340:156–162

  21. Letourneau PC, Shattuck TA, Ressler AH (1987) “Pull” and “push” in neurite elongation: observations on the effects of different concentrations of cytochalasin B and taxol. Cell Motil Cytoskelet 8:193–209

  22. Littleton JT, Bellen HJ, Perin MS (1993) Expression of synaptotagmin in Drosophila reveals transport and localization of synaptic vesicles to the synapse. Development 118:1077–1088

  23. Morris CE, Homann U (2001) Cell surface area regulation and membrane tension. J Membr Biol 179:79–102

  24. Osen-Sand A, Catsicas M, Staple JK, Jones KA, Ayala G, Knowles J, Grenningloh G, Catsicas S (1993) Inhibition of axonal growth by SNAP-25 antisense oligonucleotides in vitro and in vivo. Nature 364:445–448

  25. Pedersen SF, Hoffmann EK, Mills JW (2001) The cytoskeleton and cell volume regulation. Comp Biochem Physiol Part A Mol Integr Physiol 130:385–399

  26. Pfenninger KH, Johnson MP (1983) Membrane biogenesis in the sprouting neuron. I. Selective transfer of newly synthesized phospholipid into the growing neurite. J Cell Biol 97:1038–1042

  27. Pfenninger KH, Maylie-Pfenninger MF (1981a) Lectin labeling of sprouting neurons. I. Regional distribution of surface glycoconjugates. J Cell Biol 89:536–546

  28. Pfenninger KH, Maylie-Pfenninger MF (1981b) Lectin labeling of sprouting neurons. II. Relative movement and appearance of glycoconjugates during plasmalemmal expansion. J Cell Biol 89:547–559

  29. Pfenninger KH, de la Hossaye BA, Frame L, Helmke S, Lockerbie RO, Lohse K, Miller V, Negre-Aminou P, Wood MR (1992) Biochemical dissection of plasmalemmal expansion at the growth cone. In: Letourneau PC, Kater SB, Macagno ER, Letourneau PC, Kater SB, Macagno E (eds) The nerve growth cone. Raven Press, New York, pp 111–123

  30. Pfister BJ, Iwata A, Meaney DF, Smith DH (2004) Extreme stretch growth of integrated axons. J Neurosci 24:7978–7983

  31. Pipa RL, Woolever PS (1965) Insect neurometamorphosis. II. The fine structure of perineurial connective tissue, adipohemocytes and the shortening ventral nerve cord of a moth, Galleria mellonella. Z Zellfforsch 68:80–101

  32. Popov S (2001) Axonal transport and dynamics of microtubules. Biol Membr 18:446–452

  33. Popov S, Brown A, Poo MM (1993) Forward plasma membrane flow in growing nerve processes. Science 259:244–246

  34. Rossler W, Oland LA, Higgins MR, Hildebrand JG, Tolbert LP (1999) Development of a glia-rich axon-sorting zone in the olfactory pathway of the moth Manduca sexta. J Neurosci 19:9865–9877

  35. Shirasu M, Kimura K, Kataoka M, Takahashi M, Okajima S, Kawaguchi S, Hirasawa Y, Ide C, Mizoguchi A (2000) VAMP-2 promotes neurite elongation and SNAP-25A increases neurite sprouting in PC12 cells. Neurosci Res 37:265–275

  36. Siwicki KK, Ladewski L (2003) Associative learning and memory in Drosophila: beyond olfactory conditioning. Behav Processes 64:225–238

  37. Sollner TH (2003) Regulated exocytosis and SNARE function (Review). Mol Membr Biol 20:209–220

  38. Strausfeld NJ, Seyan HS (1985) Convergence of visual, haltere, and prosternal inputs at neck motor neurons of Calliphora erythrocephala. Cell Tissue Res 247:5–10

  39. Strausfeld NJ, Hansen L, Li YS, Gomez RS, Ito K (1998) Evolution, discovery, and interpretations of arthropod mushroom bodies. Learn Mem 5:11–37

  40. Strausfeld NJ, Sinakevitch I, Vilinsky I (2003) The mushroom bodies of Drosophila melanogaster: an immunocytological and Golgi study of Kenyon cell organization in the calyces and lobes. Microsc Res Tech 62:151–169

  41. Vance JE, Campenot RB, Vance DE (2000) The synthesis and transport of lipids for axonal growth and nerve regeneration. Bba-Mol Cell Biol L 1486:84–96

  42. Vilinsky I, Stewart BA, Drummond J, Robinson I, Deitcher DL (2002) A Drosophila SNAP-25 null mutant reveals context-dependent redundancy with SNAP-24 in neurotransmission. Genetics 162:259–271

  43. Vogt L, Giger RJ, Ziegler U, Kunz B, Buchstaller A, Hermens W, Kaplitt MG, Rosenfeld MR, Pfaff DW, Verhaagen J, Sonderegger P (1996) Continuous renewal of the axonal pathway sensor apparatus by insertion of new sensor molecules into the growth cone membrane. Curr Biol 6:1153–1158

  44. Weiss P (1941) Nerve patterns: the mechanics of nerve growth. Growth, Third Growth Symposium 5:163–203

  45. Wilkinson GS, Dodson GN (1997) Function and evolution of antlers and eye stalks in flies. In: Choe JB, Crespi B (ed) The evolution of mating systems in insects and arachnids. Chambridge University Press, Chambridge, pp 310–328

  46. Yoshihara M, Adolfsen B, Littleton JT (2003) Is synaptotagmin the calcium sensor? Curr Opin Neurobiol 13:315–323

  47. Zakharenko S, Popov S (1998) Dynamics of axonal microtubules regulate the topology of new membrane insertion into the growing neurites. J Cell Biol 143:1077–1086

  48. Zakharenko S, Popov S (2000) Plasma membrane recycling and flow in growing neurites. Neuroscience 97:185–194

  49. Zheng J, Lamoureux P, Santiago V, Dennerll T, Buxbaum RE, Heidemann SR (1991) Tensile regulation of axonal elongation and initiation. J Neurosci 11:1117–1125

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We would like to thank Dr. Gerald S. Wilkinson for generously providing flies and much help on rearing them. Drs. Hugo Bellen and David Deitcher kindly provided us with antibodies. Dr. David Deitcher provided much help with the Western blot. We thank Drs. Pat Rivlin, Ilya Vilinsky, and Heather Hoy for comments on the manuscript and Brandon Loveall for technical assistance.

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Correspondence to Elke K. Buschbeck.

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This research is supported by the National Science Foundation (IBN-9974512 and IBN-211770).

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Buschbeck, E.K., Hoy, R.R. The development of a long, coiled, optic nerve in the stalk-eyed fly Cyrtodiopsis whitei. Cell Tissue Res 321, 491–504 (2005). https://doi.org/10.1007/s00441-005-1142-4

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  • Nerve growth
  • Axon elongation
  • Eyestalk
  • Glia
  • Cyrtodiopsis whitei (Insecta)