Posterior Lateral Line Nerve Research Articles

Macrophages enhance regeneration of lateral line neuromast derived from interneuromast cells through TGF-β in zebrafish.

Macrophages play a pivotal role in the response to injury, contributing significantly to the repair and regrowth of damaged tissues. The external lateral line system in aquatic organisms offers a practical model for studying regeneration, featuring interneuromast cells connecting sensory neuromasts. Under normal conditions, these cells remain dormant, but their transformation into neuromasts occurs when overcoming inhibitory signals from Schwann cells and posterior lateral line nerves. The mechanism enabling interneuromast cells to evade inhibition by Schwann cells remains unclear. Previous observations suggest that macrophages physically interact with neuromasts, nerves, and Schwann cells during regeneration. This interaction leads to the regeneration of neuromasts in a subset of zebrafish with ablated neuromasts. To explore whether macrophages achieve this effect through secreted cytokines, we conducted experiments involving tail amputation in zebrafish larvae and tested the impact of cytokine inhibitors on neuromast regeneration. Most injured larvae remarkably regenerated a neuromast within 4 days post-amputation. Intriguingly, removal of macrophages and inhibition of the anti-inflammatory cytokine transforming growth factor-beta (TGF-β) significantly delayed neuromast regeneration. Conversely, inhibition of the pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) had minor effects on the regeneration process. This study provides insights into how macrophages activate interneuromast cells, elucidating the pathways underlying neuromast regeneration.

Timely Schwann cell division drives peripheral myelination in vivo via the laminin/cAMP pathway.

Schwann cells (SCs) migrate along peripheral axons and divide intensively to generate the right number of cells prior to axonal ensheathment; however, little is known regarding the temporal and molecular control of their division and its impact on myelination. We report that Sil, a spindle pole protein associated with autosomal recessive primary microcephaly, is required for temporal mitotic exit of SCs. In sil-deficient cassiopeia (csp-/-) mutants, SCs fail to radially sort and myelinate peripheral axons. Elevation of cAMP, but not Rac1 activity, in csp-/- restores myelin ensheathment. Most importantly, we show a significant decrease in laminin expression within csp-/- posterior lateral line nerve and that forcing Laminin 2 expression in csp-/- fully restores the ability of SCs to myelinate. Thus, we demonstrate an essential role for timely SC division in mediating laminin expression to orchestrate radial sorting and peripheral myelination in vivo.

The Lateral Line System in the Nurseryfish Kurtus gulliveri (Percomorpha: Kurtidae): A Distribution and Innervation of Superficial Neuromasts Unique within Percomorphs

The lateral line system and its innervation were examined in the Nurseryfish Kurtus gulliveri (family Kurtidae). The system is characterized by ca. 373,000 superficial neuromasts (SNs; at 152 mm standard length) occurring over one side of the entire body surface, including the dorsal, anal, and caudal fins. A fine-grid pattern of SNs is spread regularly over the surface, comprising many longitudinal and transverse SN rows, with the axis of best physiological sensibility perpendicular to the long axis of the neuromasts. On the head, rami of the anterior lateral line nerve are similar to those in typical percomorphs, each innervating numerous SNs by extensive ramification. On the trunk, 22 elongated dorsal ramules of the lateral ramus (of the posterior lateral line nerve) supply SNs on the dorsal half of the trunk and dorsal fin, and the dorsal longitudinal collector nerve innervates 22 canal neuromasts along the short (incomplete) trunk lateral line canal. Sixteen ventral ramules arise from the lateral ramus to supply SNs on the ventral half of the trunk and anal fin, which is similar to the pattern of the dorsal ramules. Innervation of the trunk and fins is distinctive, differing from all percomorphs known thus far.

Current Advances in Comprehending Dynamics of Regenerating Axons and Axon-Glia Interactions after Peripheral Nerve Injury in Zebrafish.

Following an injury, axons of both the central nervous system (CNS) and peripheral nervous system (PNS) degenerate through a coordinated and genetically conserved mechanism known as Wallerian degeneration (WD). Unlike central axons, severed peripheral axons have a higher capacity to regenerate and reinnervate their original targets, mainly because of the favorable environment that they inhabit and the presence of different cell types. Even though many aspects of regeneration in peripheral nerves have been studied, there is still a lack of understanding regarding the dynamics of axonal degeneration and regeneration, mostly due to the inherent limitations of most animal models. In this scenario, the use of zebrafish (Danio rerio) larvae combined with time-lapse microscopy currently offers a unique experimental opportunity to monitor the dynamics of the regenerative process in the PNS in vivo. This review summarizes the current knowledge and advances made in understanding the dynamics of the regenerative process of PNS axons. By using different tools available in zebrafish such as electroablation of the posterior lateral line nerve (pLLn), and laser-mediated transection of motor and sensory axons followed by time-lapse microscopy, researchers are beginning to unravel the complexity of the spatiotemporal interactions among different cell types during the regenerative process. Thus, understanding the cellular and molecular mechanisms underlying the degeneration and regeneration of peripheral nerves will open new avenues in the treatment of acute nerve trauma or chronic conditions such as neurodegenerative diseases.

Oligodendrocyte Neurofascin Independently Regulates Both Myelin Targeting and Sheath Growth in the CNS.

SummarySelection of the correct targets for myelination and regulation of myelin sheath growth are essential for central nervous system (CNS) formation and function. Through a genetic screen in zebrafish and complementary analyses in mice, we find that loss of oligodendrocyte Neurofascin leads to mistargeting of myelin to cell bodies, without affecting targeting to axons. In addition, loss of Neurofascin reduces CNS myelination by impairing myelin sheath growth. Time-lapse imaging reveals that the distinct myelinating processes of individual oligodendrocytes can engage in target selection and sheath growth at the same time and that Neurofascin concomitantly regulates targeting and growth. Disruption to Caspr, the neuronal binding partner of oligodendrocyte Neurofascin, also impairs myelin sheath growth, likely reflecting its association in an adhesion complex at the axon-glial interface with Neurofascin. Caspr does not, however, affect myelin targeting, further indicating that Neurofascin independently regulates distinct aspects of CNS myelination by individual oligodendrocytes in vivo.

Recurrent de novo MAPK8IP3 variants cause neurological phenotypes.

c-Jun-amino-terminal kinase-interacting protein 3 (JIP3), encoded by MAPK8IP3, is an adaptor protein of the kinesin-1 complex and essential for axonal transport in neurons. However, an association between MAPK8IP3 variants and human disease has not been established. We identified 5 individuals from four families with recurrent de novo variants c.1732C>T (p.Arg578Cys) and c.3436C>T (p.Arg1146Cys) in MAPK8IP3. The core phenotype includes spastic diplegia, intellectual disability, cerebral atrophy, and corpus callosum hypoplasia. Zebrafish embryos overexpressing human mutant JIP3 showed axon varicosities of the posterior lateral line nerve, suggesting an adverse effect on the developing axons. Our results suggest that MAPK8IP3 variants cause a neurodevelopmental disease. ANN NEUROL 2019;85:927-933.

Transmission Electron Microscopy for Zebrafish Larvae and Adult Lateral Line Nerve.

Transmission electron microscopy (TEM) enables visualization of the ultrastructure of the myelin sheath. Schwann cells on the posterior lateral line nerves and motor nerves can be imaged by TEM. Here, we detail the multiday processing of larval trunks and dissected posterior lateral line for TEM, as well as how to trim embedded samples, section, and stain grids for imaging.

Mechanosensory organ regeneration in zebrafish depends on a population of multipotent progenitor cells kept latent by Schwann cells.

BackgroundRegenerating damaged tissue is a complex process, requiring progenitor cells that must be stimulated to undergo proliferation, differentiation and, often, migratory behaviors and morphological changes. Multiple cell types, both resident within the damaged tissue and recruited to the lesion site, have been shown to participate. However, the cellular and molecular mechanisms involved in the activation of progenitor cell proliferation and differentiation after injury, and their regulation by different cells types, are not fully understood. The zebrafish lateral line is a suitable system to study regeneration because most of its components are fully restored after damage. The posterior lateral line (PLL) is a mechanosensory system that develops embryonically and is initially composed of seven to eight neuromasts distributed along the trunk and tail, connected by a continuous stripe of interneuromastic cells (INCs). The INCs remain in a quiescent state owing to the presence of underlying Schwann cells. They become activated during development to form intercalary neuromasts. However, no studies have described if INCs can participate in a regenerative event, for example, after the total loss of a neuromast.ResultsWe used electroablation in transgenic larvae expressing fluorescent proteins in PLL components to completely ablate single neuromasts in larvae and adult fish. This injury results in discontinuity of the INCs, Schwann cells, and the PLL nerve. In vivo imaging showed that the INCs fill the gap left after the injury and can regenerate a new neuromast in the injury zone. Further, a single INC is able to divide and form all cell types in a regenerated neuromast and, during this process, it transiently expresses the sox2 gene, a neural progenitor cell marker. We demonstrate a critical role for Schwann cells as negative regulators of INC proliferation and neuromast regeneration, and that this inhibitory property is completely dependent on active ErbB signaling.ConclusionsThe potential to regenerate a neuromast after damage requires that progenitor cells (INCs) be temporarily released from an inhibitory signal produced by nearby Schwann cells. This simple yet highly effective two-component niche offers the animal robust mechanisms for organ growth and regeneration, which can be sustained throughout life.Electronic supplementary materialThe online version of this article (doi:10.1186/s12915-016-0249-2) contains supplementary material, which is available to authorized users.

Deficiency in the mRNA export mediator Gle1 impairs Schwann cell development in the zebrafish embryo

GLE1 mutations cause lethal congenital contracture syndrome 1 (LCCS1), a severe autosomal recessive fetal motor neuron disease, and more recently have been associated with amyotrophic lateral sclerosis (ALS). The gene encodes a highly conserved protein with an essential role in mRNA export. The mechanism linking Gle1 function to motor neuron degeneration in humans has not been elucidated, but increasing evidence implicates abnormal RNA processing as a key event in the pathogenesis of several motor neuron diseases. Homozygous gle1−/− mutant zebrafish display various aspects of LCCS, showing severe developmental abnormalities including motor neuron arborization defects and embryonic lethality. A previous gene expression study on spinal cord from LCCS fetuses indicated that oligodendrocyte dysfunction may be an important factor in LCCS. We therefore set out to investigate the development of myelinating glia in gle1−/− mutant zebrafish embryos. While expression of myelin basic protein (mbp) in hindbrain oligodendrocytes appeared relatively normal, our studies revealed a prominent defect in Schwann cell precursor proliferation and differentiation in the posterior lateral line nerve. Other genes mutated in LCCS have important roles in Schwann cell development, thereby suggesting that Schwann cell deficits may be a common factor in LCCS pathogenesis. These findings illustrate the potential importance of glial cells such as myelinating Schwann cells in motor neuron diseases linked to RNA processing defects.

Fighting cichlids size up opposition with lateral line

When sizing each other up before a confrontation, most males depend on all of their senses to learn more about their opponent – listening, watching and sniffing out strengths and weaknesses. However, Karen Maruska and Julie Butler, from Louisiana State University, USA, explain that fish also explore their surroundings with an additional sense: the lateral line system responds to the ebb and flow of water surrounding the fish's body. Could fish also use this ‘touch at a distance’ system to learn more about a foe? Maruska and Butler explain that fish adversaries often waft water around while posturing in preparation for an encounter and add, ‘The lateral line is poised to provide crucial information for opponent assessment’. However, no one had ever tested whether fish resort to the specialised sense in preparation for combat. ‘Astatotilapia burtoni males use a variety of aggressive behaviors, such as lateral displays, border fights, biting and mouth fights to defend their spawning territory,’ says Butler, so the scientists selected this aggressive species to test their theory.First, the duo investigated the structure of the feisty males’ lateral line system and clearly saw the sensory units that detect disturbances in water distributed on the fish's head and along the side of the body. Then they selected dominant males that were keen to defend their territory and deactivated the animals’ lateral line systems by immersing them in a solution of cobalt chloride and severing the posterior lateral line nerve. Next, they paired combinations of males, with and without active lateral line systems, in a duel over new territory and filmed the encounters.Butler explains that for each contest there were three possible outcomes: the fish did not fight; one fish capitulated to the other before full hostilities began; or the fish battled for supremacy. Analysing the outcomes of each confrontation, the duo quickly saw that the lateral line system was an essential component of a successful non-confrontational outcome.Almost 75% of the pairs where both fish had an intact lateral line system battled. However, when a dueller with a lateral line system was pitched against an opponent that had lost the sense, the duo saw a large increase in the proportion of fish that backed down before a confrontation – up from 5% in two males with intact lateral line systems to 30% where one combatant had lost the use of the lateral line. ‘This decreased motivation was likely due to an impaired ability to adequately assess the opponent’, say Butler and Maruska, adding that the disabled fish spent more time in close proximity to their opponents than the intact fish and that fish that had lost the lateral line system failed to instigate combat with the fin-flaring display that intact fish perform before a skirmish. And when Butler and Maruska analysed the battle manoeuvres, they saw that the fish with a functional lateral line system avoided contact with their opponents, while the fish that had lost the system bit and rammed each other more.Butler and Maruska were impressed to find that the fish use the additional sense to avoid damaging confrontation and they say, ‘To our knowledge, this is the first study to implicate the lateral line system as a mode of social communication necessary for assessment during agonistic interactions’.

Zebrafish prion protein PrP2 controls collective migration process during lateral line sensory system development.

Prion protein is involved in severe neurodegenerative disorders but its physiological role is still in debate due to an absence of major developmental defects in knockout mice. Previous reports in zebrafish indicate that the two prion genes, PrP1 and PrP2, are both involved in several steps of embryonic development thus providing a unique route to discover prion protein function. Here we investigate the role of PrP2 during development of a mechano-sensory system, the posterior lateral line, using morpholino knockdown and PrP2 targeted inactivation. We confirm the efficiency of the translation blocking morpholino at the protein level. Development of the posterior lateral line is altered in PrP2 morphants, including nerve axonal outgrowth and primordium migration defects. Reduced neuromast deposition was observed in PrP2 morphants as well as in PrP2−/− mutants. Rosette formation defects were observed in PrP2 morphants, strongly suggesting an abnormal primordium organization and reflecting loss of cell cohesion during migration of the primordium. In addition, the adherens junction proteins, E-cadherin and ß-catenin, were mis-localized after reduction of PrP2 expression and thus contribute to the primordium disorganization. Consequently, hair cell differentiation and number were affected and this resulted in reduced functional neuromasts. At later developmental stages, myelination of the posterior lateral line nerve was altered. Altogether, our study reports an essential role of PrP2 in collective migration process of the primordium and in neuromast formation, further implicating a role for prion protein in cell adhesion.

The lateral line system and its innervation in Zenarchopterus dunckeri (Beloniformes: Exocoetoidei: Zenarchopteridae): an example of adaptation to surface feeding in fishes

Examination of the lateral line system and its innervation in a freshwater halfbeak, Zenarchopterus dunckeri (Beloniformes: Exocoetoidei: Zenarchopteridae), revealed adaptations of a surface insect feeder. Superficial neuromasts (SNs) occurred on the head (ventral surface of lower jaw, snout, cheek, supratemporal region and opercle), trunk (forming four longitudinal rows: middorsal, dorsolateral, lateral and ventrolateral rows) and caudal fin (two rows). The middorsal row was broad, involving three longitudinal scale series (middorsal and left and right adjacent series) anteriorly on the trunk and including 809 SNs (counted on the left side); up to 108 SNs formed a more or less crescentic patch on each scale. The supratemporal ramus of the posterior lateral line nerve (PLLN) extended strongly posterior to innervate the middorsal row. The lateral ramus (of PLLN) gave off an anterior dorsal ramule innervating the dorsolateral row, four short ventral ramules innervating the lateral row, and 15 long ventral ramules innervating SNs of the ventrolateral row and canal neuromasts of the trunk lateral line.

Axon-Schwann cell interactions during peripheral nerve regeneration in zebrafish larvae.

BackgroundPeripheral nerve injuries can severely affect the way that animals perceive signals from the surrounding environment. While damage to peripheral axons generally has a better outcome than injuries to central nervous system axons, it is currently unknown how neurons re-establish their target innervations to recover function after injury, and how accessory cells contribute to this task. Here we use a simple technique to create reproducible and localized injury in the posterior lateral line (pLL) nerve of zebrafish and follow the fate of both neurons and Schwann cells.ResultsUsing pLL single axon labeling by transient transgene expression, as well as transplantation of glial precursor cells in zebrafish larvae, we individualize different components in this system and characterize their cellular behaviors during the regenerative process. Neurectomy is followed by loss of Schwann cell differentiation markers that is reverted after nerve regrowth. We show that reinnervation of lateral line hair cells in neuromasts during pLL nerve regeneration is a highly dynamic process with promiscuous yet non-random target recognition. Furthermore, Schwann cells are required for directional extension and fasciculation of the regenerating nerve. We provide evidence that these cells and regrowing axons are mutually dependant during early stages of nerve regeneration in the pLL. The role of ErbB signaling in this context is also explored.ConclusionThe accessibility of the pLL nerve and the availability of transgenic lines that label this structure and their synaptic targets provides an outstanding in vivo model to study the different events associated with axonal extension, target reinnervation, and the complex cellular interactions between glial cells and injured axons during nerve regeneration.

The trunk lateral line system and its innervation in Mugil cephalus (Mugilidae: Mugiliformes)

The trunk lateral line system and its innervation were examined in Mugil cephalus. Almost every scale had an open longitudinal groove that housed a superficial neuromast (SN), the total number of SNs being 550 (including 55 on the caudal fin). Grooved lateral line scales constituted 14 longitudinal rows (14 lateral lines), although each groove was independent, not forming a continuous common groove along adjacent scales. The lateral ramus of the posterior lateral line nerve gave off four characteristic ramules: the anteriormost two dorsal ramules extended strongly backward, innervating SNs on the dorsal surface of the trunk; the anteriormost ventral ramule was extensively ramified to innervate SNs on the anterior flank, passing ventrally in front of the pectoral fin base; and the fifth ventral ramule extended posteriorly close to the anal fin origin, innervating SNs on the ventral half of the middle flank. The remaining SNs were innervated by 29 dorsal, 33 lateral and 25 ventral ramules, excepting those on the caudal fin. The innervation pattern revealed that the sixth lateral line (from uppermost) was homologous with that typically found in teleosts, the others being considered as additional. The sixth lateral line was anteriorly regressive, suggesting that the high insertion of the pectoral fin on the trunk is a secondary condition.

Dynamics of degeneration and regeneration in developing zebrafish peripheral axons reveals a requirement for extrinsic cell types

BackgroundUnderstanding the cellular mechanisms regulating axon degeneration and regeneration is crucial for developing treatments for nerve injury and neurodegenerative disease. In neurons, axon degeneration is distinct from cell body death and often precedes or is associated with the onset of disease symptoms. In the peripheral nervous system of both vertebrates and invertebrates, after degeneration of detached fragments, axons can often regenerate to restore function. Many studies of axonal degeneration and regeneration have used in vitro approaches, but the influence of extrinsic cell types on these processes can only be fully addressed in live animals. Because of its simplicity and superficial location, the larval zebrafish posterior lateral line (pLL) nerve is an ideal model system for live studies of axon degeneration and regeneration.ResultsWe used laser axotomy and time-lapse imaging of pLL axons to characterize the roles of leukocytes, Schwann cells and target sensory hair cells in axon degeneration and regeneration in vivo. Immune cells were essential for efficient removal of axonal debris after axotomy. Schwann cells were required for proper fasciculation and pathfinding of regenerating axons to their target cells. Intact target hair cells were not themselves required for regeneration, but chemical ablation of neuromasts caused axons to transiently deviate from their normal paths.ConclusionsMacrophages, Schwann cells, and target sensory organs are required for distinct aspects of pLL axon degeneration or regeneration in the zebrafish larva. Our work introduces a powerful vertebrate model for analyzing axonal degeneration and regeneration in the living animal and elucidating the role of extrinsic cell types in these processes.

Morphological structure and peripheral innervation of the lateral line system in the Siberian sturgeon (Acipenser baerii)

Light and scanning electron microscopy (SEM) were used to study the epidermal lateral line system of the Siberian sturgeon (Acipenser baerii Brandt, 1869). This system consists of mechanoreceptive neuromasts, ampullae and the electroreceptive organ. The neuromasts are located in 5 pairs of cephalic and 1 pair of trunk canals and superficially in the middle and posterior pit lines that lie dorsomedially along the top of the skull immediately adjacent to the otic ampullae field. Both canal neuromasts and pit organ superficial neuromasts have opposite polarized hair cells that are parallel along the axis of the canal and pit line, respectively. However, they differ in both size and shape and in the density and length of the hair bundles. The ampullae are confined on the head, adjacent to the neuromast lines. The morphological structure of the ampullae in the Siberian sturgeon is similar to the ampullae in elasmobranchs and other primitive fish. Nevertheless, it has a relatively large mucus-filled ampulla, and a shorter and narrower canal leading to a small opening to the outer epidermal surface. We also present new information concerning the peripheral innervation of lateral line receptors in sturgeons. The receptors of the lateral line system are innervated by 2 pairs of cranial nerves: anterior and posterior lateral line nerves. The peripheral processes of the anterior lateral line nerve form superficial ophthalmic, buccal, otic and anteroventral rami. The peripheral processes of the posterior lateral line nerve form middle, supratemporal and lateral rami.

Ambiguities in the identification of batoid lateral line systems clarified by innervation

Innervation of the lateral line canal system in seven batoid species (representing seven families in three orders) provided a reliable basis for the identification of each canal element. Previous topographic definitions of the canal elements have failed to recognize homologies within batoids for the scapular and hyomandibular canals. The former is innervated by the posterior lateral line nerve and the latter is innervated by an external mandibular branch of the anterior lateral line nerve. This indicates that the scapular canal is represented only by the scapular loop in Myliobatoidei.

Description and innervation of the lateral line system in two gobioids, Odontobutis obscura and Pterogobius elapoides (Teleostei: Perciformes)

Components of the lateral line system and their innervation were studied in Odontobutis obscura (Odontobutidae) and Pterogobius elapoides (Gobiidae), which are benthic and pelagic species, respectively. Innervation of the superficial neuromasts constituting the trunk lateral line system by way of three continuous longitudinal series (dorsal, middle, and ventral series: ld, lm, and lv series, respectively) became apparent for the first time. Innervation patterns indicated that the ld and lv series represented a mixture of displaced rows (from lm series) and new additional rows. In O. obscura, the ld and lv series were poorly developed, whereas both series were well developed in the pelagic P. elapoides, possibly as an adaptation to receive stimuli from above and below. Two extremely elongated nerve branches derived from the lateral ramus of the posterior lateral line nerve innervated the ld and lv series, respectively, in P. elapoides. Homologies of the neuromast rows on the head and body were discussed on the basis of their innervation patterns.

Schwann cells reposition a peripheral nerve to isolate it from postembryonic remodeling of its targets

Although much is known about the initial construction of the peripheral nervous system (PNS), less well understood are the processes that maintain the position and connections of nerves during postembryonic growth. Here, we show that the posterior lateral line nerve in zebrafish initially grows in the epidermis and then rapidly transitions across the epidermal basement membrane into the subepidermal space. Our experiments indicate that Schwann cells, which myelinate axons in the PNS, are required to reposition the nerve. In mutants lacking Schwann cells, the nerve is mislocalized and the axons remain in the epidermis. Transplanting wild-type Schwann cells into these mutants rescues the position of the nerve. Analysis of chimeric embryos suggests that the process of nerve relocalization involves two discrete steps - the degradation and recreation of the epidermal basement membrane. Although the outgrowth of axons is normal in mutants lacking Schwann cells, the nerve becomes severely disorganized at later stages. In wild-type embryos, exclusion of the nerve from the epidermis isolates axons from migration of their targets (sensory neuromasts) within the epidermis. Without Schwann cells, axons remain within the epidermis and are dragged along with the migrating neuromasts. Our analysis of the posterior lateral line system defines a new process in which Schwann cells relocate a nerve beneath the epidermal basement membrane to insulate axons from the postembryonic remodeling of their targets.

A vesicular glutamate transporter in lampreys: cDNA cloning and early expression in the nervous system

Vesicular glutamate transporters (VGLUTs) accumulate glutamate into synaptic vesicles of glutamatergic neurons, and thus are considered to define the phenotype of these neurons. Glutamate also appears to play a role in the development of the nervous system of vertebrates. Here we report the characterization of a vesicular glutamate transporter of lamprey (lVGluT), a novel member of the VGluT gene family. Phylogenetic analysis indicates that lVGLUT cannot be assigned to any of the three VGLUT isoforms characterized in teleosts and mammals, suggesting that these classes may have been fixed after the splitting between cyclostomes and gnathostomes. Expression pattern analysis during lamprey embryogenesis and prolarval stages shows that lVGluT expression is restricted to the nervous system. The first structure to express lVGluT was the olfactory epithelium of late embryos. In the brain of early prolarvae, lVGluT was expressed in most of the neuronal populations that generate the early axonal scaffold. lVGluT expression was also observed in neuronal populations of the rhombencephalon and spinal cord and in ganglia of the branchiomeric, octaval and posterior lateral line nerves. In the rhombencephalon, lVGluT expression appears to be spatially restricted in dorsal and ventral longitudinal domains. Comparison of the early expression of VGluT genes between the lamprey and some anamniotan gnathostomes (frog, zebrafish) reveals a conserved expression pattern, likely to reflect ancestral vertebrate characteristics.