Formation of the peripheral nervous system during tail regeneration in urodele amphibians: ultrastructural and immunohistochemical studies of the origin of the cells.

In the regenerating newt tail, epimorphic regeneration--which recapitulates morphologically normal embryonic development--proceeds along a rostrocaudal differentiation gradient. Innervation of the new myomeres results from the spinal roots of segments rostral to the amputation plane and from ventral roots emerging from the lateroventral region of the regenerating spinal cord, in which motor neurons are differentiating. Electron microscopy and an indirect immunofluorescence study with anti-glial fibrillary acid protein (GFAP) confirm that the ventrolateral part of the regenerated ependymal tube gives rise to cells of the ventral root sheath and the spinal ganglia. Anti-GFAP and anti-neurofilament antibodies showed that ependymoglial cells and Schwann cells may play a role in neuronal pathfinding by helping guide and stabilize pioneering axons as they extend toward the myomeres. The carbohydrate epitope NC-1 is expressed in the spinal cord, in sheath cells of the spinal ganglia and in the non-myelin-forming Schwann cells of the peripheral nervous system. L1, a Ca++ independent neural cell adhesion molecule, was detected in the axonal compartments of the regenerating spinal cord, on immature and/or non-myelin-forming Schwann cells within the peripheral nervous system (PNS), and on nerve fibers within the regenerate. These immunohistochemical observations collectively support the hypothesis that Schwann cells already present in the blastema could be involved in organizing neural pathways.     

Embryology of the thoracic and abdominal ganglia of the large milkweed bug, Oncopeltus fasciatus (Dallas), (Hemiptera, Lygaeidae)

In this study, the condensation of the three thoracic and 11 abdominal segmental ganglia to form a prothoracic and central nerve mass during embryogenesis is described. During katatrepsis, many changes occur in the organization of these ganglia; this study suggests that some of these changes are caused by mechanical forces acting on the ventral nerve cord at this time. The ventral nerve cord begins its anterior migration and coalescence ten hours after katatrepsis and is completed 63 hours later. The central ganglion is made up of the meso- and metathoracic ganglia and seven abdominal ganglia. Intrasegmental median cord nuclei are shown to form glial elements in the median sagittal plane of the neuropile and in the longitudinal connectives. Intersegmental median cord neuroblasts migrate into the posterior gangliomeres but, apparently, degenerate soon after katatrepsis. Lateral cord cells bordering on the neuropile form a glial investment that surrounds this fiber tract region. Peripheral lateral cord cells are shown to form the cells of the outer ganglionic sheath, the perineurium.     

Fine structure of a lepidopteran nervous system and its accessibility to peroxidase and lanthanum

Summary The avascular ventral nerve cord of the moth, Manduca sexta, possesses an extensive dorsal mass of connective tissue in which lie fibroblasts that produce a collagen-like protein. The lateral and ventral surfaces of the nerve cord are ensheathed by an acellular neural lamella. Beneath this lies a layer of microtubule-laden perineurial cells which are separated from one another at their peripheral borders by lacunae containing electron-opaque material to which the cells are attached by hemi-desmosomes. Beyond these spaces, narrow intercellular clefts occur between the interdigitating perineurial plasma membranes; these are then connected by both gap and tight junctions. The axons beneath are surrounded by glia which also contain many microtubules and which are linked to one another by desmosomes and tight junctions.When intact nerve cords are incubated in horseradish peroxidase, reaction product is subsequently found within the neural lamella as well as in the lacunae and clefts between perineurial cells, but not beyond this level. Desheathed preparations, however, contain peroxidase within the cytoplasm of the exposed glial cells. Lanthanum penetrates the neural lamella and the lacunae, clefts and gap junctions between adjacent perineurial cells, but no further. It therefore appears that the tight junctions in the perineurium may be the site of restriction to the entry of ions and molecules, the existence of which has been suggested previously by electrophysiological investigations.I am grateful to Miss Yvonne R. Carter for her invaluable technical assistance and to Dr. J.E. Treherne and Dr. D.B. Sattelle for helpful discussions.     

The neural tube origin of ventral root sheath cells in the chick embryo

The embryonic origin of peripheral nerve Schwann/sheath cells is still uncertain. Although the neural crest is known to be an important source, it is not clear whether the ventral neural tube also contributes a progenitor population for motor axons. We have used the techniques of immunohistochemistry, electron microscopy and quail-chick grafting to examine this problem. Immunohistochemistry with monoclonal antibody HNK-1 identified a cluster of immunoreactive cells in the sclerotome, at the site of the future ventral root. With the electron microscope, nucleated cells could not be seen breaching the basal lamina of the neural tube, exclusively in the region of the ventral root and preceding axon outgrowth. After grafting a length of crest-ablated quail neural tube in place of host chick neural tube, a population of quail cells was found localized to the ventral root exit zone, associated with the ventral root axons. Taken together, these observations support the possibility of a neural tube origin for ventral root sheath cells, although we found no evidence for a more extensive migration of these cells. The ventral root cells share certain phenotypic traits, such as HNK-1 immunoreactivity, with neural-crest-derived Schwann cells, but are not necessarily identical to them. We argue that while they may help motor axons to exit the neural tube at the correct position, they are unlikely to guide axons beyond the immediate vicinity of the neural tube.     

Ultrastructure studies of the extracellular filaments in the ventral nerve of Panulirus argus

The ventral nerve cord of the spiny lobster, Panulirus argus was examined by transmission and scanning electron microscopy. Tannic acid mordant stain was used to enhance extracellular filaments. The ventral nerve cord is surrounded by an unusual perineurial sheath composed primarily of interwoven extracellular filaments. Gap junctions were found associated with the glial cells making up the perineurium. The axo-glial wrappings also contained extracellular filaments associated in bundles rather than uniformly around the axons. The extracellular filaments of the perineurium and axo-glial wrappings appeared to be morphologically identical with diameters ranging from 10-15 nm.     

Immunohistochemical localization of gastrin-cholecystokinin-like material in the central nervous system of the migratory locust

Summary Brain, corpora cardiaca (CC)-corpora allata (CA) complex, suboesophageal ganglion, thoracic and abdominal ganglia of adults, larvae and embryos of Locusta migratoria have been immunohistochemically screened for gastrin cholecystokinin (CCK-8(s))-like material. In adult, numerous immunoreactive neurons and nerve fibres are located, with a marked symmetry, in various parts of the brain and throughout the ventral nerve cord. In the median part of the brain, cell bodies belonging neither to cellular type A1 nor A2 (following Victoria blue-paraldehyde fuchsin staining) are immunopositive; their processes terminate in the upper protocerebral neuropile. In lateral parts of the brain, external cell bodies send axons into CC and some up to CA, other internal have processes which terminate in the neuropile of the brain. Two of these latter cells react also with methionine-enkephalin antiserum. In the ventral nerve cord, in addition to numerous perikarya, immunore-active arborizations terminate in the neuropile or in close association with the sheath, at the dorsal part of all ganglia.This CCK-8(s) distribution pattern is observed only at the two last larval instars, but is precociously detected in the abdominal nerve cord of embryos, one day before hatching.     

The midline glial cells are required for regionalization of commissural axons in the embryonic CNS of Drosophila

 The ventral nerve cord of arthropods is characterised by the organisation of major axon tracts in a ladder-like pattern. The individual neuromeres are connected by longitudinal connectives whereas the contra-lateral connections are brought about through segmental commissures. In each neuromere of the embryonic central nervous system (CNS) of Drosophila an anterior and a posterior commissure is found. The development of these commissures requires a set of neurone-glia interactions at the midline. Here we show that both the anterior as well as the posterior commissures are subdivided into three axon-containing regions. Electron microscopy of the ventral nerve cord of mutations affecting CNS midline cells indicates that the midline glial cells are required for this subdivision. In addition the midline glial cells appear required for a crossing of commissural growth cones perpendicular to the longitudinal tracts, since in mutants with defective midline glial cells commissural axons frequently cross the midline at aberrant angles. Received: 6 July 1997 / Accepted: 27 August 1997     

Ultrastructural distribution of cholinesterase activity in the ventral nerve cord of the earthworm Lumbricus terrestris

Summary The fine structure and distribution of cholinesterase (ChE) activity in the ventral nerve cord of the earthworm (Lumbricus terrestris) was studied, using acetyl- and butyrylthiocholine iodides as substrates and iso-OMPA, 284C51 and eserine as inhibitors to discriminate between acetylcholinesterase (AChE) and other cholinesterase (ns.ChE) activities.The earthworm ventral nerve cord exhibits intense ChE activity. Both AChE and ns.ChE were present and they had identical distribution, being located mainly in the supportive glial cells. Most neurones of the ventral nerve cord contained no histochemically demonstrable activity. The ventral giant nerve cells were observed with the electron microscope to exhibit AChE activity. The enzyme was situated in the membranes of the rough-surfaced endoplasmic reticulum and in peculiar lamellated bodies but not in the membranes of the Golgi complex.     

On the neurosecretory system of Dendrobaena atheca Cernosvitov

Four kinds of neurosecretory cells A, B, U and C are distinguished in the central nervous system of Dendrobaena atheca Cernosvitov. A cells, which show different morphological characteristics under different physiological states and during their cyclic changes, are the most active neurosecretory cells. They form the outer layer of the cortical cell zone in the cerebral ganglion. B cells are large and medium sized and are distributed in all parts of the central nervous system. U cells are found only in the sub-pharyngeal ganglion while C cells are distributed in the sub-pharyngeal as well as in the ventral nerve cord ganglion. The number and secretory activity of C cells decrease in caudal direction. Further, Gomori-positive cells are also observed in the ganglia of the vegetative nervous system. A rudimentary neurohaemal organ, the storage zone, has been observed in the cerebral ganglion and there appears to be another neurohaemal area in the ventral nerve cord ganglion. The storage zone is formed by the terminal ends of the axons of A cells. The chrome alum haematoxylin phloxin (CHP) and aldehyde fuchsin (AF) positive substances in the form of granules are found in this area. The cerebral ganglion is richly supplied by blood capillaries. The distal end of the axons of B cells are swollen like a bulb while in some cases the axons are united to form an axonal tract. Extra-cellular material is abundant in different parts of the nervous system. In all cell types, the perinuclear zone is the first to show activity in the secretory cycle. It appears that the nucleus may be involved in the elaboration of the neurosecretory material in the cells.     

The nervous system ofNectonema munidae andGordius aquaticus,with implications for the ground pattern of the Nematomorpha

 The nervous system of Nectonema munida is shown to be composed of a brain, a ventral nerve cord with an anterior and a posterior enlargement, a dorsal nerve cord and a plexus-like basiepidermal nervous system. The ultrastructure of these parts is given. Additionally, the ventral nerve cord of Gordius aquaticus is ultrastructurally described. The results are compared with the literature to work out the ground pattern of the Nematomorpha according to the nervous system. This contains a circumpharyngeal brain with a main subpharyngeal portion and a weak suprapharyngeal portion, a ventral and dorsal intraepidermal nerve cord and a peripheral nervous system. The ground pattern of the nervous system of Nematomorpha is then compared to that of other Nemathelminthes. The form of the brain and the distribution of perikarya are derived characters of the Nematomorpha. The existence of an unpaired ventral and an unpaired dorsal nerve cord and the position of these two cords in epidermal cords are synapomorphies of the Nematomorpha and the Nematoda. Accepted: 7 July 1996     

Ultrastructural and immunocytochemical observations of the nervous systems of three macrodasyidan gastrotrichs

The nervous systems of three macrodasyidan gastrotrichs, Dactylopodola baltica, Macrodasys caudatus and Dolichodasys elongatus, were investigated using immunocytochemistry and electron microscopy. Labelling of neural structures against serotonin revealed the presence of two pairs of cerebral cells, a dorsal cerebral connective, and paired ventral nerve cords in D. baltica. In M. caudatus and D. elongatus serotonin immunoreactivity was present in a single pair of dorsal cerebral cells and the ventral nerve cords; the dorsal connective of D. elongatus was also immunoreactive to serotonin and acetylated α‐tubulin. The presence of paired, serotonin‐like immunoreactive cells in D. baltica and other species may represent the plesiomorphic condition in Macrodasyida. The fine structure of the photoreceptors in D. baltica was also investigated to explore the potential ground pattern for eyes in the Macrodasyida. The pigmented photoreceptors of D. baltica contain a unicellular pigment cup, sheath cell and sensory receptor. The pigment cup contains numerous osmiophilic granules that presumably function to shield the eyes from downwelling light in the red part of the spectrum. Projecting into the pigment cup and sheath cell are numerous microvilli from a bipolar sensory cell. A single sensory cell may represent the plesiomorphic condition in Macrodasyida, with multiplication of sensory cells representative of more derived taxa.     

Two regions in the isolated brainstem of the frog that modulate respiratory-related activity

Using microinjection techniques, we have explored the isolated, complete midline sectioned brainstem of the frog (Rana catesbeiana) to identify regions that influence the endogenous respiratory-related motor activity. Ten-nanoliter injections of lidocaine (1%), GABA (100 mM) and glutamate (10 and 100 mM) into discrete regions of the rostral and the caudal brainstem produced different effects on the phasic neural discharge. In the rostral site lidocaine, GABA and glutamate injections altered neural burst frequency with little or no effect on burst amplitude. In the caudal site, responses to lidocaine and GABA injections consisted primarily of decreases in neural burst amplitude, often, but not always associated with minor decreases in burst frequency. In this same region, the response to glutamate was characterized by a temporary interruption of the rhythmic neural burst activity. The largest responses to substance injection in both regions were obtained at sites ranging between 200 and 500 m from the ventral surface, in the ventral medullary reticular formation. The results reveal the existence of two areas in the frog brainstem that influence respiratory motor output, one related to the respiratory burst frequency and the other related to the amplitude of the motor output.Abbreviations V trigeminal nerve - VI abducens nerve - VII facial nerve - VIII auditory nerve - X vagal nerve - H hypoglossal nerve - VRG ventral respiratory group - NTS nucleus of the solitary tract     

Competence of blastomeres for the expression of molecular tissue markers is acquired by diverse mechanisms in the embryo of Platynereis (Annelida)

This paper is devoted to the role of cell divisions for the establishment of histospecificity in the embryo of the spiralian, Platynereis dumerilii (Annelida). We have incubated successive cleavage stages in cytochalasin B (CCB) and observed whether the cells thereafter were able to acquire the competence for expressing histospecific antigens of larval gland cells (labelled by the monoclonal antibody OI64) and of neural components of the ventral nerve cord (labelled by mAb OI7 or by testing acety1cholinesterase activity), respectively. Incubation in CCB results in permanent cleavage arrest, but does not necessarily interfere with biochemical differentiation of such markers. Synthesis of the differentiation marker specific for larval gland cells does not require any cleavages but this capacity becomes restricted to the 1a and 1b cell lines if cleavages are allowed to occur. In contrast, the progenitors of neural cells need at least 6.5 h of normal development before they acquire the competence to synthesize two neural differentiation markers, which can be demonstrated after at least two more days of development. Thus, prespecifying and diversifying cleavages are a prerequisite for neurogenesis and production of the investigated neural markers. Competence for the expression of histospecific antigens may also depend on cell-cell interactions. If 20–24 h old embryos are treated with puromycin, pioneering fibres fail to grow out from a pair of posterior nerve cell progenitors as they would have done normally 24–48 h after fertilization. Concomitantly, a number of potential nerve cells which now do not come into contact with pioneering fibres do not express the neural antigen. This suggests that a local inductive stimulus from the pioneering fibres normally imprints cell fate onto ventral plate cells and turns them into neuroblasts. Correspondence to: A.W.C. Dorresteijn     

The distribution of pheromone-biosynthesis-activating neuropeptide (PBAN) immunoreactivity in the central nervous system of the corn earworm moth,Helicoverpa zea

Summary Production of sex pheromone in several species of moths has been shown to be under the control of a neuropeptide termed pheromone-biosynthesis-activating neuropeptide (PBAN). We have produced an antiserum to PBAN from Helicoverpa zea (Lepidoptera: Noctuidae) and used it to investigate the distribution of immunoreactive peptide in the brain-suboesophageal ganglion complex and its associated neurohemal structures, and the segmental ganglia of the ventral nerve cord. Immunocytochemical methods reveal three clusters of cells along the ventral midline in the suboesophageal ganglion (SOG), one cluster each in the presumptive mandibular (4 cells), maxillary (12–14 cells), and labial neuromeres (4 cells). The proximal neurites of these cells are similar in their dorsal and lateral patterns of projection, indicating a serial homology among the three clusters. Members of the mandibular and maxillary clusters have axons projecting into the maxillary nerve, while two additional pairs of axons from the maxillary cluster project into the ventral nerve cord. Members of the labial cluster project to the retrocerebral complex (corpora cardiaca and cephalic aorta) via the nervus corpus cardiaci III (NCC III). The axons projecting into the ventral nerve cord appear to arborize principally in the dorsolateral region of each segmental ganglion; the terminal abdominal ganglion is distinct in containing an additional ventromedial arborization in the posterior third of the ganglion. Quantification of the extractable immunoreactive peptide in the retrocerebral complex by ELISA indicates that PBAN is gradually depleted during the scotophase, then restored to maximal levels in the photophase. Taken together, our findings provide anatomical evidence for both neurohormonal release of PBAN as well as axonal transport via the ventral nerve cord to release sites within the segmental ganglia.Abbreviations A aorta - Br-SOG brain-suboesophageal ganglion complex - CC corpus cardiacum - PBS phosphate-buffered saline - PLI PBAN-like immunoreactivity - TAG terminal abdominal ganglion - VNC ventral nerve cord     

Galleria mellonella nerve cords in vitro: stage-specific survival and differential responsiveness to beta-ecdysone

The survival of ventral nerve cord segments of Galleria mellonella in tissue culture medium, ascertained by histological and biochemical criteria, ability to shorten when transplanted, and responsiveness to β-ecdysone, is correlated with the stage of development of the donor, and with the presence of a connective tissue sheath, the neural lamella, about the segment. After only 18 hr in vitro connectives which have lost the neural lamella during metamorphosis no longer have the capacity to shorten when transplanted or when exposed to β-ecdysone in the culture medium. By contrast, after 7 days in vitro connectives with a neural lamella shorten appreciably when β-ecdysone is added, or when they are exposed to the humoral milieu of a host undergoing metamorphosis. β-Ecdysone stimulates the incorporation of uridine-5-³H in segments both with and without the neural lamella, but only in segments which have previously begun their metamorphosis. Since shortening in response to β-ecdysone occurs only in connectives which have already begun to shorten, β-ecdysone appears to accelerate physiological processes underway before it is added to the medium rather than initiate metamorphosis in cultured nerve cords.     

Peripheral nerve connections influence the appearance of neurosecretary material in neural sheath of ventral ganglion of the fly Sarcophaga bullata: an immunocytochemical study

This study examined the role of the brain and peripheral connections with the target organs in the appearance of neurosecretary material within the dorsal neural sheath of the ventral ganglion of the fly S. bullata. Specifically, the accumulation of the neuropeptide FMRFamide (the neurosecretary material) was examined by immunocytochemistry. Immunoreactions were performed on: (1) a normal intact ventral ganglion, (2) an isolated ventral ganglion that was cultured in vivo, and (3) a ventral ganglion that was isolated by transection from the brain, but retained its peripheral nerve connections. The results demonstrate that (a) the neurons of the ganglia survive and exhibit FMRFamide immune reaction independent of their peripheral connections, and (b) the accumulation of neuropeptide in the dorsal neural sheath is controlled by intact peripheral nerve connections with the ganglion. It is suggested that in the absence of their peripheral connections, the axons of FMRFamide immunoreactive neurons fail to invade the neural sheath resulting in the accumulation of neurosecretary material.     

The relationship between the ventral nerve cord,body size and phylogeny in ground beetles (Coleoptera: Carabidae)

The ventral nerve cord in the family Carabidae (considered in the widest sense! exhibits variations in the degree of fusion of thoracic and abdominal ganglia. There are usually three discrete thoracic ganglia and between one and seven discrete abdominal ganglia, the number differing between tribes. Of the 44 tribes and 177 species examined, 38 tribes contained species showing no differences in the degree of ventral nerve cord consolidation. However, the remaining six tribes showed variations in the degree of ventral nerve cord consolidation between genera (Lebiini, Cychrini, Nebriini, Scaritini, Licinini and Brachinini), whilst one genus showed variations between species (Leistus, Nebriini). No variation in ventral nerve cord consolidation was observed in conspecifics. The degree of ventral nerve cord consolidation is inversely proportional to overall body length. With respect to phylogeny, the degree of consolidation of the nerve cord docs not consistently support the traditional Carabinae-Harpalinae subfamily division. However, the Harpalinae always have four or less discrete abdominal ganglia (with the sole exception of the Broscinij, whilst the Carabinae exhibit almost the whole range of variations. Thus the Harpalinae (or the major pari of it) may be a monophyletic group, but this is not true of the Carabinae. Trends in the degree of ventral nerve cord consolidation for the various tribes were noted, and phylogenetic implications were evaluated wherever possible.     

Multisegmental cobalt filling of the dorsal giant fibers in the nervous system of the earthworm,Lumbricus terrestris

Summary The anatomical organization of the two dorsal giant fiber systems of the earthworm Lumbricus terrestris is demonstrated in whole mounts and serial-section reconstructions based on backfillings of the ventral nerve cord with cobalt chloride. Both the medial and lateral fiber systems can be labeled selectively over more than ten body segments. They show a characteristic segmental pattern of collaterals with some modification in tail segments and of dorsal plasma protrusions in the unpaired medial giant fiber presumably representing openings in the myelin sheath. We found no multisegmental cobalt transport in other large neurons of the nerve cord. Cobalt passes through the segmentai septa between consecutive axonal elements of the metameric giant fibers and presumably also through commissural contacts between specific collaterals of the lateral giant fibers. Since these sites of contact are known to represent electrical synapses, cobalt coupling may, in L. terrestris, correlate with functional electrotonic coupling.Abbreviations CL collateral of lateral giant fiber - CM collateral of medial giant fiber - GIN giant interneuron - LGF lateral giant fiber - MGF medial giant fiber - SN segmental nerve     

The whole-body shortening reflex of the medicinal leech: motor pattern,sensory basis,and interneuronal pathways

The leech whole-body shortening reflex consists of a rapid contraction of the body elicited by a mechanical stimulus to the anterior of the animal. We used a variety of reduced preparations — semi-intact, body wall, and isolated nerve cord — to begin to elucidate the neural basis of this reflex in the medicinal leech Hirudo medicinalis. The motor pattern of the reflex involved an activation of excitatory motor neurons innervating dorsal and ventral longitudinal muscles (dorsal excitors and ventral excitors respectively), as well as the L cell, a motor neuron innervating both dorsal and ventral longitudinal muscles. The sensory input for the reflex was provided primarily by the T (touch) and P (pressure) types of identified mechanosensory neuron. The S cell network, a set of electrically-coupled interneurons which makes up a fast conducting pathway in the leech nerve cord, was active during shortening and accounted for the shortest-latency excitation of the L cells. Other, parallel, interneuronal pathways contributed to shortening as well. The whole-body shortening reflex was shown to be distinct from the previously described local shortening behavior of the leech in its sensory threshold, motor pattern, and (at least partially) in its interneuronal basis.Abbreviations conn connective - DE dorsal excitor motor neuron - DI dorsal inhibitor motor neuron - DP dorsal posterior nerve - DP:B1 dorsal posterior nerve branch 1 - DP:B2 dorsal posterior nerve branch 2 - MG midbody ganglion - VE ventral excitor motor neuron - VI ventral inhibitor motor neuron     

Conserved and novel functions for Netrin in the formation of the axonal scaffold and glial sheath cells in spiders

Netrins are well known for their function as long-range chemotropic guidance cues, in particular in the ventral midline of vertebrates and invertebrates. Over the past years, publications are accumulating that support an additional short-range function for Netrins in diverse developmental processes such as axonal pathfinding and cell adhesion. We describe here the formation of the axonal scaffold in the spiders Cupiennius salei and Achaearanea tepidariorum and show that axonal tract formation seems to follow the same sequence as in insects and crustaceans in both species. First, segmental neuropiles are established which then become connected by the longitudinal fascicles. Interestingly, the commissures are established at the same time as the longitudinal tracts despite the large gap between the corresponding hemi-neuromeres which results from the lateral movement of the germband halves during spider embryogenesis. We show that Netrin has a conserved function in the ventral midline in commissural axon guidance. This function is retained by an adaptation of the expression pattern to the specific morphology of the spider embryo. Furthermore, we demonstrate a novel function of netrin in the formation of glial sheath cells that has an impact on neural precursor differentiation. Loss of Netrin function leads to the absence of glial sheath cells which in turn results in premature segregation of neural precursors and overexpression of the early motor- and interneuronal marker islet. We suggest that Netrin is required in the differentiated sheath cells for establishing and maintaining the interaction between NPGs and sheath cells. This short-range adhesive interaction ensures that the neural precursors maintain their epithelial character and remain attached to the NPGs. Both the conserved and novel functions of Netrin seem to be required for the proper formation of the axonal scaffold.