Pioneer neurones use basal lamina as a substratum for outgrowth in the embryonic grasshopper limb

Development ◽  
1988 ◽  
Vol 104 (4) ◽  
pp. 601-608 ◽  
Author(s):  
H. Anderson ◽  
R.P. Tucker
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Basal Lamina ◽  
Growth Cone ◽  
Growth Cones ◽  
Axon Outgrowth ◽  
Stages Of Development

During axonogenesis, contacts made by the growth cone with its substratum are important in guiding the direction of neurone outgrowth. This study examines the contacts made by the growth cones of pioneer neurones in the embryonic grasshopper limb. Individual pioneer neurones at different stages of development were injected with horseradish peroxidase and the contacts made by the filopodia at the tip of their growth cones were examined by electron microscopy. Filopodia made few contacts with mesodermal cells, some contacts with ectodermal cells and very frequent contacts with basal lamina underlying the ectoderm. Components of the basal lamina may therefore play a role in guiding pioneer axon outgrowth.

Growth cones stall and collapse during axon outgrowth in Caenorhabditis elegans

Development ◽  
1999 ◽  
Vol 126 (20) ◽  
pp. 4489-4498 ◽  
Author(s):  
K.M. Knobel ◽  
E.M. Jorgensen ◽  
M.J. Bastiani
Keyword(s):  
Caenorhabditis Elegans ◽  
Growth Cone ◽  
Nerve Cord ◽  
Body Wall ◽  
Growth Cones ◽  
The Body ◽  
Axon Outgrowth ◽  
Attachment Structures ◽  
Dorsal Nerve

During nervous system development, neurons form synaptic contacts with distant target cells. These connections are formed by the extension of axonal processes along predetermined pathways. Axon outgrowth is directed by growth cones located at the tips of these neuronal processes. Although the behavior of growth cones has been well-characterized in vitro, it is difficult to observe growth cones in vivo. We have observed motor neuron growth cones migrating in living Caenorhabditis elegans larvae using time-lapse confocal microscopy. Specifically, we observed the VD motor neurons extend axons from the ventral to dorsal nerve cord during the L2 stage. The growth cones of these neurons are round and migrate rapidly across the epidermis if they are unobstructed. When they contact axons of the lateral nerve fascicles, growth cones stall and spread out along the fascicle to form anvil-shaped structures. After pausing for a few minutes, they extend lamellipodia beyond the fascicle and resume migration toward the dorsal nerve cord. Growth cones stall again when they contact the body wall muscles. These muscles are tightly attached to the epidermis by narrowly spaced circumferential attachment structures. Stalled growth cones extend fingers dorsally between these hypodermal attachment structures. When a single finger has projected through the body wall muscle quadrant, the growth cone located on the ventral side of the muscle collapses and a new growth cone forms at the dorsal tip of the predominating finger. Thus, we observe that complete growth cone collapse occurs in vivo and not just in culture assays. In contrast to studies indicating that collapse occurs upon contact with repulsive substrata, collapse of the VD growth cones may result from an intrinsic signal that serves to maintain growth cone primacy and conserve cellular material.

A scanning electron microscope study of the development of a peripheral sensory neurite network

Development ◽  
1982 ◽  
Vol 69 (1) ◽  
pp. 237-250
Author(s):  
Alan Roberts ◽  
J. S. H. Taylor
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Basal Lamina ◽  
Growth Cone ◽  
Growth Cones ◽  
Sensory Innervation ◽  
Scanning Electron Microscope Study ◽  
Scanning Electron

The formation of the sensory neurite plexus on the basal lamina of trunk skin in Xenopus embryos has been examined using the scanning electron microscope. It is formed by Rohon-Beard and extramedullary neurons which provide the first sensory innervation of the skin. By observing the distribution of growth cones on the inside surface of the skin of embryos at different ages, the development of the plexus has been followed and related to the development of sensitivity to sensory stimulation. The general features of the plexus are illustrated using a photomontage taken at × 1100. Measurements on neurites from this, and of growth cone orientations demonstrate a general ventral growth pattern with some small regional variations. Interactions of neurites within the plexus are examined. Neurites meeting at shallow angles tend to fasciculate, whilethose meeting at close to 90° tend to cross each other. Angles of incidence and separation of neurites show few angles less than 30°, which suggests that active adjustments occur after a growth cone meets or leaves another neurite. The observations allow comparison of behaviour of growing neurites in vivo and in vitro. Our evidence suggests that adhesion between growth cones and neurites is stronger than that between growth cones and the basal lamina of the skin.

scholarly journals A pre-embedding immunolabeling technique for basal lamina and extracellular matrix molecules.

1988 ◽  
Vol 36 (4) ◽  
pp. 453-458 ◽  
Author(s):  
M M Martins-Green ◽  
K T Tokuyasu
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Extracellular Matrix ◽  
Optical Microscopy ◽  
Basal Lamina ◽  
Serial Sections ◽  
Stages Of Development ◽  
Additional Advantage ◽  
Transmission Electron ◽  
Extracellular Matrix Molecules

We have developed a pre-embedding immunolabeling technique to identify basal lamina and extracellular matrix molecules in embryos at various stages of development. The technique works for both fluorescence optical microscopy (1-2.5-micron sections) and for transmission electron microscopy, and enables straigthforward correlation between the two. An additional advantage is the easy preparation of well-oriented serial sections, facilitating detailed studies of development.

scholarly journals The C. elegans gene vab-8 guides posteriorly directed axon outgrowth and cell migration

Development ◽  
1996 ◽  
Vol 122 (2) ◽  
pp. 671-682 ◽  
Author(s):  
B. Wightman ◽  
S.G. Clark ◽  
A.M. Taskar ◽  
W.C. Forrester ◽  
A.V. Maricq ◽  
...  
Keyword(s):  
Nervous System ◽  
Cell Migration ◽  
Growth Cone ◽  
Growth Cones ◽  
Body Axis ◽  
Axon Outgrowth ◽  
Common Mechanism ◽  
C Elegans

The assembly of the nervous system in the nematode C. elegans requires the directed migrations of cells and growth comes along the anteroposterior and dorsoventral body axis. We show here that the gene vab-8 is essential for most posteriorly directed migrations of cells and growth cones. Mutations in vab-8 disrupt fourteen of seventeen posteriorly directed migrations, but only two of seventeen anteriorly directed and dorsoventral migrations. For two types of neurons that extend axons both anteriorly and posteriorly, vab-8 mutations disrupt only the growth of the posteriorly directed axon. vab-8 encodes two genetic activities that function in the guidance of different migrations. Our results suggest that most posteriorly directed cell and growth cone migrations are guided by a common mechanism involving the vab-8 gene.

scholarly journals A specific Ret receptor isoform is required for pioneer axon outgrowth and growth cone dynamics

2018 ◽  
Author(s):  
Adam M. Tuttle ◽  
Catherine M. Drerup ◽  
Molly H. Marra ◽  
Alex V. Nechiporuk
Keyword(s):  
Growth Cone ◽  
Axon Growth ◽  
Retrograde Transport ◽  
Growth Cones ◽  
Axon Outgrowth ◽  
Loss Of Function ◽  
Anterograde Transport ◽  
Interacting Protein ◽  
Pioneer Neurons

AbstractIn many cases, axon growth and guidance are driven by pioneer axons, the first axons to grow in a particular region. Despite their dynamic pathfinding capabilities and developmental importance, there are very few pioneer neuron specific markers and thus their in vivo identification and functional interrogation have been difficult. We found that a Ret receptor isoform, Ret51, is highly enriched in peripheral sensory pioneer neurons and is required for pioneer axon outgrowth. Ret null mutant pioneer neurons differentiate normally; however, they displayed defects in growth cone morphology and formation of filopodia before pioneer axon extension prematurely halts. We also demonstrate loss-of-function of a retrograde cargo adaptor, JNK-interacting protein 3 (Jip3), phenocopied many of these axonal defects. We further found that loss of Jip3 led to accumulation of activated Ret receptor in pioneer growth cones, indicating a failure in the clearance of activated Ret from growth cones. Using an axon sever approach as well as in vivo analysis of axonal transport, we showed Jip3 specifically mediates retrograde, but not anterograde, transport of activated Ret51. Finally, live imaging revealed that Jip3 and Ret51 were retrogradely co-transported in pioneer axons, suggesting Jip3 functions as an adapter for retrograde transport of Ret51. Taken together, these results identify Ret51 as a molecular marker of pioneer neurons and elucidate an important isoform-specific role for Ret51 in axon growth and growth cone dynamics during development.

Patterns of peanut agglutinin binding within the developing grasshopper central nervous system

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 49-56
Author(s):  
Richard Burt ◽  
Hilary Anderson
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Horseradish Peroxidase ◽  
Neurite Outgrowth ◽  
Binding Sites ◽  
Axon Outgrowth ◽  
Peanut Agglutinin ◽  
Stages Of Development ◽  
Early Stages ◽  
Neuronal Processes

The location of peanut agglutinin (PNA) binding was investigated in the segmental ganglia of the developing grasshopper embryo. Neuronal processes were stained but cell bodies were not. The first appearance of PNA binding in development was associated with the first neurons to initiate axon outgrowth, the progeny of the MP2 cells. In the early stages of development the location of PNA binding was congruent with that of antibodies against horseradish peroxidase (HRP), which bind to neurons. In more advanced ganglia only a subpopulation of those neuronal processes that bound auti-HRP also bound PNA. The results suggest that PNA-binding sites are present only on those neuronal processes which are still developing and raise the possibility that these molecules may play a rôle in neurite outgrowth and navigation.

scholarly journals Neuronal growth cones respond to laser-induced axonal damage

2011 ◽  
Vol 9 (68) ◽  
pp. 535-547 ◽  
Author(s):  
Tao Wu ◽  
Samarendra Mohanty ◽  
Veronica Gomez-Godinez ◽  
Linda Z. Shi ◽  
Lih-Huei Liaw ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Growth Cone ◽  
Axon Growth ◽  
Growth Cones ◽  
Damage Site ◽  
Transmission Electron ◽  
Fluorescence And Electron Microscopy ◽  
Cone Response ◽  
Laser Induced Damage

Although it is well known that damage to neurons results in release of substances that inhibit axonal growth, release of chemical signals from damaged axons that attract axon growth cones has not been observed. In this study, a 532 nm 12 ns laser was focused to a diffraction-limited spot to produce site-specific damage to single goldfish axons in vitro . The axons underwent a localized decrease in thickness (‘thinning’) within seconds. Analysis by fluorescence and transmission electron microscopy indicated that there was no gross rupture of the cell membrane. Mitochondrial transport along the axonal cytoskeleton immediately stopped at the damage site, but recovered over several minutes. Within seconds of damage nearby growth cones extended filopodia towards the injury and were often observed to contact the damaged site. Turning of the growth cone towards the injured axon also was observed. Repair of the laser-induced damage was evidenced by recovery of the axon thickness as well as restoration of mitochondrial movement. We describe a new process of growth cone response to damaged axons. This has been possible through the interface of optics (laser subcellular surgery), fluorescence and electron microscopy, and a goldfish retinal ganglion cell culture model.

Comparative strengths and weaknesses of peroxidase and colloidal gold in pre- and post-embedding immunolabeling techniques

1987 ◽  
Vol 45 ◽  
pp. 1002-1005
Author(s):  
D. R. Abrahamson ◽  
P. L. St.John ◽  
E. W. Perry
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Light Microscopy ◽  
Colloidal Gold ◽  
Light Microscope ◽  
Ultrastructural Localization ◽  
The Other ◽  
Quantitative Studies ◽  
Dense Suspension ◽  
Antigen Localization

Antibodies coupled to tracers for electron microscopy have been instrumental in the ultrastructural localization of antigens within cells and tissues. Among the most popular tracers are horseradish peroxidase (HRP), an enzyme that yields an osmiophilic reaction product, and colloidal gold, an electron dense suspension of particles. Some advantages of IgG-HRP conjugates are that they are readily synthesized, relatively small, and the immunolabeling obtained in a given experiment can be evaluated in the light microscope. In contrast, colloidal gold conjugates are available in different size ranges and multiple labeling as well as quantitative studies can therefore be undertaken through particle counting. On the other hand, gold conjugates are generally larger than those of HRP but usually can not be visualized with light microscopy. Concern has been raised, however, that HRP reaction product, which is exquisitely sensitive when generated properly, may in some cases distribute to sites distant from the original binding of the conjugate and therefore result in spurious antigen localization.

Ultrastructural study of the avian ventral lateral geniculate nucleus

1991 ◽  
Vol 6 (2) ◽  
pp. 119-134 ◽  
Author(s):  
Gloria D. Guiloff
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Lateral Geniculate Nucleus ◽  
Ultrastructural Study ◽  
Lateral Geniculate

AbstractThe ultrastructure of the pigeon and quail ventral lateral geniculate nucleus was analyzed with standard electron microscopy and horseradish peroxidase tracing of its retinal and tectal afferents. Six types of neurons were distinguished: two large, two medium-sized, and two small types.

scholarly journals Regulated Actin Cytoskeleton Assembly at Filopodium Tips Controls Their Extension and Retraction

1999 ◽  
Vol 146 (5) ◽  
pp. 1097-1106 ◽  
Author(s):  
Aneil Mallavarapu ◽  
Tim Mitchison
Keyword(s):  
Actin Cytoskeleton ◽  
Growth Cone ◽  
Actin Filaments ◽  
Neuroblastoma Cell ◽  
Initial Step ◽  
Growth Cones ◽  
Neuroblastoma Cell Line ◽  
Dominant Factor ◽  
Retrograde Flow ◽  
Assembly Rate

The extension and retraction of filopodia in response to extracellular cues is thought to be an important initial step that determines the direction of growth cone advance. We sought to understand how the dynamic behavior of the actin cytoskeleton is regulated to produce extension or retraction. By observing the movement of fiduciary marks on actin filaments in growth cones of a neuroblastoma cell line, we found that filopodium extension and retraction are governed by a balance between the rate of actin cytoskeleton assembly at the tip and retrograde flow. Both assembly and flow rate can vary with time in a single filopodium and between filopodia in a single growth cone. Regulation of assembly rate is the dominant factor in controlling filopodia behavior in our system.

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