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Ultrastructure of an identified array of growth cones and possible substrates for guidance in the embryonic medicinal leech, Hirudo medicinalis

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Abstract

The oblique muscle organizer (Comb- or C-cell) in the embryonic medicinal leech, Hirudo medicinalis, provides an amenable situation to examine growth cone navigation in vivo. Each of the segmentally iterated C-cells extends an array of growth cones through the body wall along oblique trajectories. C-cell growth cones undergo an early, relatively slow period of extension followed by later, protracted and rapid directed outgrowth. During such transitions in extension, guidance might be mediated by a number of factors, including intrinsic constraints on polarity, spatially and temporally regulated cell and matrix interactions, physical constraints imposed by the environment, or guidance along particular cells in advance of the growth cones. Growth cones and their environment were examined by transmission electron microscopy to define those factors that might play a significant role in migration and guidance in this system. The ultrastructural examination has made the possibility very unlikely that simple, physical constraints play a prominent role in guiding C-cell growth cones. No anatomically defined paths or obliquely aligned channels were found in advance of these growth cones, and there were no identifiable physical boundaries, which might constrain young growth cones to a particular location in the body wall before rapid extension. There were diverse associations with many matrices and basement membranes located above, below, and within the layer in which growth cones appear to extend at the light level. Additionally, a preliminary examination of myocyte assembly upon processes proximal to the growth cones further implicates a role for matrix-associated interactions in muscle histogenesis as well as process outgrowth during embryonic development.

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References

  • Acklin SE, Nicholls JG (1990) Intrinsic and extrinsic factors influencing properties and growth patterns of identified leech neurons in culture. J Neurosci 10:1082–1090

    Google Scholar 

  • Anderson H, Tucker RP (1988) Pioneer neurones use basal lamina as a substratum for outgrowth in the embryonic grasshopper limb. Development 104:601–608

    Google Scholar 

  • Argiro V, Bunge MB, Johnson MI (1984) Correlation between growth form and movement and their dependence on neuronal age. J Neurosci 4:3051–3062

    Google Scholar 

  • Ball EE, Ho RK, Goodman CS (1985) Development of neuromuscular specificity in the grasshopper embryo: guidance of motoneuron growth cones by muscle pioneers. J Neurosci 5:1808–1819

    Google Scholar 

  • Bastiani MJ, Goodman CS (1984) Neuronal growth cones: specific interactions mediated by filopodial insertion and induction of coated vesicles. Proc Natl Acad Sci USA 81:1849–1853

    Google Scholar 

  • Bentley D, O'Connor TP (1992) Guidance and steering of peripheral pioneer growth cones in grasshopper embryos. In: PC Letourneau, SB Kater, ER Macagno (eds) The nerve growth cone. Raven Press, New York, pp 265–282

    Google Scholar 

  • Bovolenta P, Dodd J (1990) Guidance of commissural growth cones at the floor plate in embryonic rat spinal cord. Development 109:435–447

    Google Scholar 

  • Bovolenta P, Mason C (1987) Growth cone morphology varies with position in the developing mouse visual pathway from retina to first targets. J Neurosci 7:1447–1460

    Google Scholar 

  • Bray DF, Wagenaar EB (1978) A double staining technique for improved contrast of thin sections from Spurr-embedded tissue. Can J Bot 56:129–132

    Google Scholar 

  • Bunge MB (1973) Fine structure of nerve fibers and growth cones of isolated sympathetic neurons in culture. J Cell Biol 56:713–735

    Google Scholar 

  • Byers HR, Fujiwara K (1982) Stress fibers in cells in situ: immunofluorescence visualization with anti-actin, anti-myosin and anti-alpha-actinin. J Cell Biol 93:808–811

    Google Scholar 

  • Caudy M, Bentley D (1986 a) Pioneer growth cone morphologies reveal proximal increases in substrate affinity within leg segments of grasshopper embryos. J Neurosci 6:364–379

    Google Scholar 

  • Caudy M, Bentley D (1986 b) Pioneer growth cone steering along a series of neuronal and non-neuronal cues of different affinities. J Neurosci 6:1781–1795

    Google Scholar 

  • Chiquet M, Acklin SE (1986) Attachment to Con A or extracellular matrix initiates rapid sprouting by cultured leech neurons. Proc Natl Acad Sci USA 83:6188–6192

    Google Scholar 

  • Chiquet M, Masuda-Nakagawa L, Beck K (1988) Attachment to an endogenous laminin-like protein initiates sprouting by leech neurons. J Cell Biol 107:1189–1198

    Google Scholar 

  • Condic ML, Bentley D (1989 a) Removal of the basal lamina in vivo reveals growth cone-basal lamina adhesive interactions and axonal tension in grasshopper embryos. J Neurosci 9:2678–2686

    Google Scholar 

  • Condic ML, Bentley D (1989 b) Pioneer growth cone adhesion in vivo to boundary cells and neurons after enzymatic removal of basal lamina in grasshopper embryos. J Neurosci 9:2687–2696

    Google Scholar 

  • Dotti CG, Sullivan CA, Banker GA (1988) The establishment of polarity by hippocampal neurons in culture. J Neurosci 8:1454–1468

    Google Scholar 

  • Easter Jr. SS, Bratton B, Scherer SS (1984) Growth-related order of the retinal fiber layer in goldfish. J Neurosci 4:2173–2190

    Google Scholar 

  • Egar M, Singer M (1972) The role of ependyma in spinal cord regeneration in the urodele, Triturus. Exp Neurol 37:422–430

    Google Scholar 

  • Egar M, Singer M (1977) Interependymal channels and cell death in normal development of chick hindbrain. Anat Rec 187:573–574

    Google Scholar 

  • Egar M, Simpson SB, Singer M (1970) The growth and differentiation of the regenerating spinal cord of the lizard. J Morphol 131:131–152

    Google Scholar 

  • Fawcett DW (1966) The cell: an atlas of fine structure. Saunders, Philadelphia

    Google Scholar 

  • Fernández J, Stent GS (1982) Embryonic development of the hirudinid leech Hirudo medicinalis: structure, development and segmentation of the germinal plate. J Embryol Exp Morphol 72:71–96

    Google Scholar 

  • Gillon JW, Wallace BG (1984) Segmental variation in the arborization of identified neurons in the leech central neuvous system. J Comp Neurol 228:142–148

    Google Scholar 

  • Grumbacher-Reinert S (1989) Local influence of substrate molecules in determining distinctive growth patterns of identified neurons in culture. Proc Natl Acad Sci USA 86:7270–7274

    Google Scholar 

  • Halfter W (1989) Antisera to basal lamina and glial endfeet disturb the normal extension of axons on retina and pigment epithelium basal laminae. Development 107:281–297

    Google Scholar 

  • Halfter W, Diamantis I, Monard D (1988) Migratory behavior of cells on embryonic retina basal lamina. Dev Biol 130:259–275

    Google Scholar 

  • Harris WA, Holt CE, Bonhoeffer F (1987) Retinal axons with and without their somata, growing to and arborizing in the tectum of Xenopus embryos: a time-lapse video study of single fibers in vivo. Development 101:123–133

    Google Scholar 

  • Harrison RG (1935) On the origin and development of the nervous system studied by the methods of experimental embryology. Proc R Soc Lond [Biol] 118:155–196

    Google Scholar 

  • Heuser JE, Kirschner M (1980) Filament organization revealed in platinum replicas of freeze-dried cytoskeletons. J Cell Biol 86:212–234

    Google Scholar 

  • Ho RK, Ball EE, Goodman CS (1983) Muscle pioneers: large mesodermal cells that erect a scaffold for developing muscles and motoneurones in grasshopper embryos. Nature 301:66–69

    Google Scholar 

  • Holt CE (1989) A single-cell analysis of early retinal ganglion cell differentiation in Xenopus: from soma to axon tip. J Neurosci 9:3123–3145

    Google Scholar 

  • Jellies J (1990 a) Muscle assembly in simple systems. Trends Neurosci 13:126–131

    Google Scholar 

  • Jellies J (1990 b) Spatial and temporal regulation of sialated ganglioside (GQ)-like immunoreactivity on a stereotyped growth cone array (abstract). Soc Neurosci 16:152

    Google Scholar 

  • Jellies J, Kristan WB Jr (1988 a) Embryonic assembly of a complex muscle is directed by a single identified cell in the medicinal leech. J Neurosci 8:3317–3326

    Google Scholar 

  • Jellies J, Kristan WB Jr (1988 b) Directional cues influence, but do not determine stereotyped navigation of a growth cone array in the embryonic leech (abstract). Soc Neurosci 14:451

    Google Scholar 

  • Jellies J, Kristan WB Jr (1991) The oblique muscle organizer in Hirudo medicinalis, an identified embryonic cell projecting multiple parallel growth cones in an orderly array. Dev Biol 148:334–354

    Google Scholar 

  • Jellies J, Loer CM, Kristan WB Jr (1987) Morphological changes in leech Retzius neurons after target contact during embryogenesis. J Neurosci 7:2618–2629

    Google Scholar 

  • Jellies J, Kopp DM, Geisert EE Jr (1993) Developmental regulation of a glycolipid epitope on actively extending growth cones and central and peripheral projections in the medicinal leech. Dev Biol 159:691–705

    Google Scholar 

  • Kapfhammer JP, Raper JA (1987) Interactions between growth cones and neurites growing from different neural tissues in culture. J Neurosci 7:1595–1600

    Google Scholar 

  • Kater S, Letourneau P (1985) Biology of the nerve growth cone. Liss, New York

    Google Scholar 

  • Katz MJ, Lasek RJ (1981) Substrate pathways demonstrated by transplanted Mauthner axons. J Comp Neurol 195:627–641

    Google Scholar 

  • Katz MJ, Lasek RJ, Nauta HJ (1980) Ontogeny of substrate pathways and the origin of neural circuit pattern. Neuroscience 5:821–833

    Google Scholar 

  • Keynes R, Cook G (1990) Cell-cell repulsion: clues from the growth cone? Cell 62:609–610

    Google Scholar 

  • Kim GJ, Shatz CJ, McConnell SK (1991) Morphology of pioneer and follower growth cones in the developing cerebral cortex. J Neurobiol 22:629–642

    Google Scholar 

  • Kopp DM, Jellies J (1992) Multimorphic growth cones in the embryonic medicinal leech: relationship between shape changes and outgrowth transitions. J Comp Neurol 328:393–405

    Google Scholar 

  • Kopp DM, McCarthy D, Jellies J (1991) Ultrastructure of an identified growth cone array in embryonic leech reveals spatially diverse cell and matrix interactions during directed migration (abstract). Soc Neurosci 17:15

    Google Scholar 

  • Krayanek S, Goldberg S (1981) Oriented extracellular channels and axonal guidance in the embryonic chick retina. Dev Biol 84:41–50

    Google Scholar 

  • Letourneau PC (1979) Cell-substratum adhesion of neurite growth cones, and its role in neurite elongation. Exp Cell Res 124:127–138

    Google Scholar 

  • Letourneau PC, Condic ML, Snow DM (1992 a) Extracellular matrix and neurite outgrowth. Curr Opin Genet Dev 2:625–634

    Google Scholar 

  • Letourneau PC, Kater SB, Macagno ER (1992 b) The nerve growth cone. Raven Press, New York

    Google Scholar 

  • Masuda-Nakagawa LM, Nicholls JG (1991) Extracellular matrix molecules in development and regeneration of the leech CNS. Philos Trans R Soc Lond [Biol] 331:323–335

    Google Scholar 

  • Masuda-Nakagawa L, Beck K, Chiquet M (1988) Identification of molecules in leech extracellular matrix that promote neurite outgrowth. Proc R Soc Lond [Biol] 235:247–257

    Google Scholar 

  • Muller KJ, Carbonetto S (1979) The morphological and physiological properties of a regenerating synapse in the CNS of the leech. J Comp Neurol 185:485–516

    Google Scholar 

  • Muller KJ, Nicholls JG, Stent GS (1981) Neurobiology of the leech. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Nordlander RH (1987) Axonal growth cones in the developing amphibian spinal cord. J Comp Neurol 263:485–496

    Google Scholar 

  • Nordlander RH, Singer M (1978) The role of ependyma in regeneration of the spinal cord in the urodele amphibian tail. J Comp Neurol 180:349–374

    Google Scholar 

  • Nordlander RH, Singer M (1982 a) Spaces precede axons in Xenopus embryonic spinal cord. Exp Neurol 75:221–228

    Google Scholar 

  • Nordlander RH, Singer M (1982 b) Morphology and position of growth cones in the developing Xenopus spinal cord. Dev Brain Res 4:181–193

    Google Scholar 

  • O'Connor TP, Duerr JS, Bentley D (1990) Pioneer growth cone steering decisions mediated by single filopodial contacts in situ. J Neurosci 10:3935–3946

    Google Scholar 

  • Patterson PH (1988) On the importance of being inhibited, or saying no to growth cones. Neuron 1:263–267

    Google Scholar 

  • Raper JA, Bastiani M, Goodman CS (1983) Pathfinding by neuronal growth cones in grasshopper embryos. I. Divergent choices made by the growth cones of sibling neurons. J Neurosci 3:20–30

    Google Scholar 

  • Reichardt LF, Tomaselli KJ (1991) Extracellular matrix molecules and their receptors: functions in neural development. Annu Rev Neurosci 14:531–570

    Google Scholar 

  • Roberts A, Patton DT (1985) Growth cones and the formation of central and periperal neurites by sensory neurones in amphibian embryos. J Neurosci Res 13:23–28

    Google Scholar 

  • Silver J, Robb RM (1979) Studies on the development of the eye cup and optic nerve in normal mice and in mutants with congenital optic nerve aplasia. Dev Biol 68:175–190

    Google Scholar 

  • Silver J, Sidman RL (1980) A mechanism for the guidance and topographic patterning of retinal ganglion cell axons. J Comp Neurol 189:101–111

    Google Scholar 

  • Singer M, Nordlander RH, Egar M (1979) Axonal guidance during embryogenesis and regeneration in the spinal cord of the newt: the blueprint hypothesis of neuronal pathway patterning. J Comp Neurol 185:1–22

    Google Scholar 

  • Stuart DK, Thomson I, Weisblat DA, Kramer AP (1982) Antibody staining of embryonic leech muscle, blast cell migration and neuronal pathway formation (abstract). Soc Neurosci 8:15

    Google Scholar 

  • Taghert PH, Bastiani MJ, Ho RK, Goodman CS (1982) Guidance of pioneer growth cones: filopodial contacts and coupling revealed with an antibody to Lucifer yellow. Dev Biol 94:391–399

    Google Scholar 

  • Tennyson VM (1970) The fine structure of the axon and growth cone of the dorsal root neuroblast of the rabbit embryo. J Cell Biol 44:62–79

    Google Scholar 

  • Tomaselli KJ, Neugebauer KM (1991) 2001 interations? An extracellular space odyssey. Curr Opin Neurobiol 1:364–369

    Google Scholar 

  • Torrence SA, Stuart DK (1986) Gangliogenesis in leech embryos: migration of neuronal precursor cells. J Neurosci 6:2736–2746

    Google Scholar 

  • Tosney KW, Landmesser LT (1985) Growth cone morphology and trajectory in the lumbosacral region of the chick embryo. J Neurosci 5:2345–2358

    Google Scholar 

  • Tosney KW, Wessells NK (1983) Neuronal motility: the ultrastructure of veils and microspikes correlates with their motile activities. J Cell Sci 61:389–411

    Google Scholar 

  • Trinkaus JP (1985) Further thoughts on directional cell movement during morphogenesis. J Neurosci Res 13:1–19

    Google Scholar 

  • Turner JE, Singer M (1947 a) An electron microscopic study of the newt (Triturus viridescens) optic nerve. J Comp Neurol 156:1–18

    Google Scholar 

  • Turner JE, Singer M (1947 b) The ultrastructure of regeneration in the severed newt optic nerve. J Exp Zool 190:249–268

    Google Scholar 

  • Venable JM, Coggeshall R (1965) Simplified lead citrate stain for use in electron microscopy. J Cell Biol 25:407

    Google Scholar 

  • Weiss P (1955) Neuvous system (neurogenesis). In: BJ Willier, PA Weiss, Hamburger VS (eds): Analysis of development. Saunders, Philadelphia, pp 346–401

    Google Scholar 

  • Yaginuma H, Homma S, Kunzi R, Oppenheim RW (1991) Pathfinding by growth cones of commissural interneurons in the chick embryo spinal cord: a light and electron microscopic study. J Comp Neurol 304:78–102

    Google Scholar 

  • Yamada KM, Spooner BS, Wessells NK (1971) Ultrastructure and function of growth cones and axons of cultured nerve cells. J Cell Biol 49:614–635

    Google Scholar 

  • Zipser B, McKay R (1981) Monoclonal antibodies distinguish identifiable neurones in the leech. Nature 28:549–554

    Google Scholar 

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Kopp, D.M., Jellies, J. Ultrastructure of an identified array of growth cones and possible substrates for guidance in the embryonic medicinal leech, Hirudo medicinalis . Cell Tissue Res 276, 281–293 (1994). https://doi.org/10.1007/BF00306114

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  • DOI: https://doi.org/10.1007/BF00306114

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