Development of neurons synthesizing noradrenaline and acetylcholine in the superior cervical ganglion of the rat in vivo and in vitro.

The development of neurons synthesizing noradrenaline and acetylcholine in the superior cervical ganglion of the rat has been examined after transection of the preganglionic nerve trunk in vivo and by maintenance of whole ganglia in modified Rose chambers in vitro. Temporal changes in the activities of two neurotransmitter enzymes, tyrosine hydroxylase and choline acetyltransferase, were measured as markers for neurons synthesizing noradrenaline and acetylcholine, respectively.In vivo, there was a small increase in choline acetyltransferase activity between birth and adulthood and this was comparable to the increase seen in ganglia from 2-day-old rats maintained in vitro for 14 days in the absence of nerve growth factor. Both in vivo and in vitro, high doses of nerve growth factor (10 μg/g and 1 μg/ml, respectively) resulted in increases in both tyrosine hydroxylase activity and choline acetyltransferase activity, the effect on choline acetyltransferase activity being greater in vitro than in vivo. Similar concentrations of nerve growth factor had no effect on the choline acetyltransferase activity in cultured ganglia taken from 3-week-old rats. It is concluded that increases in choline acetyltransferase activity occur only in ganglia from newborn rats because they contain neurons that still retain flexibility of development and can synthesize acetylcholine due to the influence of factors within the ganglion or in the culture medium.In vitro, following the elimination of endogenous nerve growth factor by the addition of anti-nerve growth factor, tyrosine hydroxylase activity decreased but choline acetyltransferase activity increased in cultured ganglia. High concentrations of guanethidine (100 μg/ml) caused decreases in both tyrosine hydroxylase and choline acetyltransferase activities. Following removal of guanethidine from the medium, choline acetyltransferase activity increased but tyrosine hydroxylase activity was unchanged. When guanethidine was replaced with nerve growth factor, tyrosine hydroxylase activity increased to a greater extent than choline acetyltransferase activity.Our results are discussed in terms of the presence in the ganglion of a multipotential cell type which can differentiate into either an adrenergic or a cholinergic neuron. Differentiation along cholinergic lines is accompanied by a loss of the ability to take up catecholamines and a release from dependence on nerve growth factor.     

Biochemical and morphological characterization of sympathetic neurons grown in a chemically-defined medium

Previous studies have demonstrated that individual neurons from neonatal rat superior cervical ganglion express a mixed adrenergic-cholinergic phenotype when grown under certain tissue culture conditions.^{9,14,15,29,30} The expression of this phenotype is critically influenced by a number of undefined components present in the culture medium.^18,23,33 In the present study, we have examined whether superior cervical ganglion neurons grown on a chemically defined serum-free medium similarly develop dual transmitter expression, or if under these conditions, neurons express only those properties characteristic of their adrenergic heritage. To address this issue, we established that superior cervical ganglion neurons could be maintained in culture for extended periods on the defined medium described by Bottenstein & Sato⁴ in the absence of supporting cells. We then studied the biochemical, immunocytochemical and ultrastructural characteristics of these neurons. We found that in defined medium, superior cervical ganglion neurons continued to express, in a modified form, certain of their expected adrenergic properties, including the development of tyrosine hydroxylase and dopamine-β-hydroxylase activities, stores of endogenous norepinephrine, synaptic vesicles with dense cores and tyrosine hydroxy lase-immunoreactive staining properties. Superior cervical ganglion neurons grown on a defined medium did not, however, acquire cholinergic traits in culture. In this paper we show that choline acetyltransferase activity did not reach detectable levels; the companion paper¹³ documents that cholinergic synapses were not formed.We conclude that superior cervical ganglion neurons, grown under serum-free culture conditions, develop certain properties characteristic of adrenergic neurons and do not express a mixed adrenergic cholinergic phenotype. A companion paper¹³ describes the electrophysiological properties of these neurons and demonstrates the frequent occurrence of electrotonic synapses in these cultures.     

Differences in sensitivity to nerve growth factor of axon formation and tyrosine hydroxylase induction in cultured sympathetic neurons

Superior cervical ganglia from 2-day-old and 3-week-old rats were maintained in vitro for up to 2 weeks in the presence of a range of concentrations of nerve growth factor up to 100 micrograms/ml. Nerve fibre length and density were measured and tyrosine hydroxylase activity of these cultures assayed after various times. Ganglia were also examined for catecholamines and neuronal numbers using fluorescence histochemistry and histology respectively. In cultures maintained without nerve growth factor, or in those containing low concentrations of nerve growth factor (3 ng/ml), tyrosine hydroxylase decreased to 5-10% of the initial levels by 14 days in vitro. The presence of the high concentration of 1 microgram/ml nerve growth factor in the culture medium or the addition of such a concentration during the culture period did not prevent an initial decrease in tyrosine hydroxylase but subsequently increased the enzyme activity. The maximal effect of nerve growth factor on nerve fibre density was at low concentrations whereas its maximal effect on neuronal survival, tyrosine hydroxylase activity or nerve fibre elongation was at high concentrations. After 2 days in culture, maximum neurite production occurred in cultures containing 10 ng/ml, while maximum nerve fibre elongation and tyrosine hydroxylase activity occurred in cultures containing 100 micrograms/ml nerve growth factor. We conclude that low concentrations of nerve growth factor, as occur in plasma, cause maximum axon formation while high concentrations of nerve growth factor, as occur in effector organs, induce maximum tyrosine hydroxylase activity and cell survival. The former process may be mediated via cell surface receptors and the latter via retrograde axonal transport of nerve growth factor to the cell body, following uptake by the terminal regions of the axons.     

Development of cholinergic neurones in cultures of rat superior cervical ganglia. Role of calcium and macromolecules

Superior cervical ganglia from 3-day-old rats were grown in vitro for 14 days. Addition of an extract of rat heart to the culture medium led to no change in the activity of tyrosine hydroxylase but a doubling of the activity of choline acetyltransferase in the ganglia.To see whether this effect was due to a change in the concentration of unbound Ca²⁺, the calcium-binding capacity of extracts of adult rat heart was determined in a dialysis-binding study. Both cardiac extracts and serum lowered the levels of free calcium in the medium, but by relatively small amounts. When ganglia were grown in media containing 0, 2.5 and 6 mm Ca²⁺, there was no change in the ratio of choline acetyltransferase activity to tyrosine hydroxylase activity, although the activities of both enzymes increased at lower Ca²⁺ concentrations.When superior cervical ganglia were grown in Rose chambers under cellophane strips, the choline acetyltransferase/tyrosine hydroxylase ratio was higher than when superior cervical ganglia were loose in the medium. When a larger number of ganglia was grown under cellophane this further increased the ratio, while positioning of the ganglia so that substances could diffuse out from under the cellophane decreased the choline acetyltransferase/tyrosine hydroxylase ratio. When ganglia from 3-day-old rats were grown side by side with ganglia from 3-week-old rats, there was an even greater effect on the ratio of the two enzyme activities.We conclude that the ability of cardiac extracts to increase the ratio of choline acetyltransferase activity to tyrposine hydroxylase activity in ganglia is not due to changes in the Ca²⁺ concentration in the medium, but to the presence of a non-dialysable macromolecule. A similar factor also seems to be secreted by the ganglion itself.     

Acetylcholine metabolism by cultured neurons from rat nodose ganglia: regulation by a macromolecule from muscle-conditioned medium

Acetylcholine metabolism has been studied in primary cultures of neurons dissociated from newborn rat nodose ganglia. Neither nerve growth factor nor muscle-conditioned medium had any detectable effect on the long-term survival of these neurons, which appeared well differentiated upon phase-contrast and electron microscopic examination. [3H]Acetylcholine synthesis and accumulation by 2-3-week old nodose cultures and choline acetyltransferase activity were increased (2.0 +/- 0.15)-fold and (2.0 +/- 0.48)-fold, respectively, upon growth with muscle-conditioned medium, whereas acetylcholinesterase was decreased (1.5 +/- 0.1)-fold (means +/- SEM, n = 5-9). The same effects were observed when comparing nodose cultures grown in the presence of proteins purified from conditioned medium in four fractionation steps. This purified material had no effects on the protein content of the cultures, or on lactate dehydrogenase activity, and thus did not affect the overall growth of the cultures. We demonstrate that this factor copurifies with a factor(s) involved in the regulation of choline acetyltransferase and acetylcholinesterase in neuron cultures from newborn rat superior cervical ganglia [Swerts, Le Van Thai, Vigny and Weber (1983) Devl Biol. 100, 1-11] and from embryonic day 13 rat embryo spinal cord [Giess and Weber (1984) J. Neurosci. 4, 1442-1452]. Although the cholinergic factor from muscle-conditioned medium has not been purified to homogeneity, the data suggest that the same extracellular, macromolecular factor may be involved in the regulation of acetylcholine metabolism in derivatives from the neural crest, the neural tube and the epibranchial placodes.     

Localization and axonal transport of immunoreactive cholinergic organelles in rat motor neurons—An immunofluorescent study

Antisera produced in rabbits against pure fractions of cholinergic vesicles from Narcine brasiliensis were used to study cholinergic organelles in rat motor neurons. The indirect immunofluorescence method was used on perfusion-fixed material. The rats were surgically sympathectomized to remove sympathetic adrenergic and cholinergic nerves from the sciatic nerve. In the intact animal immunoreactive material, likely to represent cholinergic vesicles, was observed in motor endplates, identified by labelling with rhodamine-conjugated α-bungarotoxin or with subsequent acetylcholinesterase staining. The motor perikarya contained very little immunoreactive material. Non-terminal axons were virtually devoid of immunofluorescence in the intact animal. After crushing the sciatic nerve, immunoreactive material (likely to represent axonal cholinergic organelles) accumulated rapidly on both sides of the crush, indicating a rapid bidirectional transport. The transport was sensitive to local application of mitotic inhibitors.The axons which accumulated immunoreactive organelles were motor axons, as demonstrated by various procedures: (1) Cutting of ventral roots prevented accumulation of immunoreactive material in the nerve. (2) Deafferentation did not notably influence accumulations of immunoreactive material. (3) Ligated axons with immunoreactive material were acetylcholinesterase positive when identification was made on the same section; the intra-axonal distribution of immunoreactive material and acetylcholinesterase was not identical, however, and the Narcine antisera did not cross-react with bovine acetylcholinesterase in a solid phase immunoassay. (4) Most axons in ventral roots, but not in dorsal roots, accumulated strongly fluorescent immunoreactive material, while axons in dorsal roots contained weakly fluorescent material. On the other hand, substance P-like immune reactivity was present in many dorsal root axons, but only very rarely in ventral roots.It is suggested that the antisera against Narcine cholinergic vesicles can be used as a marker for cholinergic organelles in the motor neuron, and may be an important tool for studying the axonal cholinergic vesicles. It cannot, however, be used to identify cholinergic structures in unknown locations because it recognizes common antigenic determinants in transmitter organelles of other nerves e.g. adrenergic nerves. The axonal cholinergic organelles may carry important molecules, other than acetylcholine to the nerve endings.     

The immunofluorescent localization of regulatory and catalytic subunits of cyclic AMP-dependent protein kinase in neuronal and glial cell types of the central nervous system

The cellular localization of the regulatory (RI and RII) and catalytic components of the cyclic AMP-dependent protein kinase in selected areas of the central nervous system was determined using a selective and sensitive immunofluorescent technique. The neuronal and glial distribution was similar for the three subunits, however, each unit exhibited differences in the pattern of subcellular localization. Whereas the catalytic unit was confined to intranuclear loci, RI and RII were identified within both the cytoplasm and nucleus, with RII in addition showing distinct staining of the nuclear membrane. A granular distribution of RI in the cell soma further distinguished the cytological localization of these two regulatory components in this cellular compartment.The contrasting distribution of the subunits suggests possible functional differences between the regulatory and catalytic components of cyclic AMP-dependent protein kinase, which are discussed with regard to the role of this enzyme in mediating the physiological effects of cyclic AMP in the central nervous system.     

Axotomy induced changes in neuronal plasticity of sympathetic chain ganglia (SChG) neurons supplying descending colon in the pig

Sympathetic neurons are capable of extensive regeneration following axonal injury. To investigate the response to axotomy of colon-projecting neurons (CPN) localized in the porcine sympathetic chain ganglia (SChG), the retrograde Fast Blue (FB) tracer, axonal transection and double immunohistochemistry methods were applied. The CPN were localized exclusively in the lumbar SChG and displayed a predominantly catecholaminergic [i.e. Tyrosine Hydroxylase (TH)/Dopamine β Hydroxylase (DβH)] and Neuropeptide Y (NPY) positive phenotype under physiological conditions. Axotomy led to a significant decrease in TH/DβH production and a simultaneous increase in the neuropeptides Galanin (GAL) and Somatostatin (SOM), but not NPY or Vasoactive Intestinal Peptide (VIP) expression in retrogradely traced perikarya. Furthermore, the decrease in density of TH-/DβH-, VIP-, Leu⁵-Enkephalin (LENK)-, Choline Acetyltransferase (ChAT)-immunoreactive (-IR) nerve fibers occurred after axotomy. These data suggest a species-specific response to axonal damage of the CPN localized in porcine SChG. Since the SChG neurons supervise the vasculature of gut both in physiological and pathological conditions, and since pig is a more accurate animal model of human gut than a rodent (Swindle et al., 1992), these data may contribute to the understanding of the pathology of several gut illnesses, like Crohn Disease and Irritable Bowel Syndrome which commonly affect western populations.     

The simultaneous presence of neuroepithelial cells and neuroepithelial bodies in the respiratory gas bladder of the longnose gar, Lepisosteus osseus, and the spotted gar, L. oculatus

Anatomical and functional studies on the autonomic innervation as well as the location of airway receptors in the air-bladder of lepisosteids are very fragmentary. These water-breathing fishes share in common with the bichirs the presence of a glottis (not a ductus pneumaticus) opening into the esophagus. In contrast to a high concentration of neuroepithelial cells (NECs) contained in the furrowed epithelium in the lung of Polypterus, these cells are scattered as solitary cells in the glottal epithelium, and grouped to form neuroepithelial bodies (NEBs) in the mucociliated epithelium investing the main trabeculae in the air-bladder of Lepisosteus osseus and L. oculatus. The present immunohistochemical studies also demonstrated the presence of nerve fibers in the trabecular striated musculature and a possible relation to NEBs in these species, and identified immunoreactive elements of this innervation. Tyrosine hydroxylase (TH), choline acetyltransferase (ChAT), 5-HT and neuropeptide immunoreactivities were detected in the intramural nerve fibers. 5-HT and VIP immunopositive nerve fibers are apparently associated with NEBs. TH, VIP and SP immunoreactivities are also present in nerve fibers coursing in the radially arranged striated muscle surrounding the glottis and its submucosa. 5-HT positive neurons are also found in submucosal and the muscle layers of the glottis. The physiological function of the adrenergic and inhibitory innervation of the striated muscle as well as the neurochemical coding and morphology of the innervation of the NEBs are not known. Future studies are needed to provide evidence for these receptors with the capacity of chemoreceptors and/or mechanoreceptors.     

Asynchrony in the expression of guanosine 3':5'-phosphate-dependent protein kinase by clusters of Purkinje cells during the perinatal development of rat cerebellum

The early maturation of Purkinje cells was studied by immunocytochemistry in the rat cerebellum. The antiserum against guanosine 3':5'-phosphate-dependent protein kinase used in this study has been shown previously to label specifically all Purkinje cells in the adult rat. Immunoreactive Purkinje cells are first observed at embryonic day 17, 2 days after the end of proliferation of this neuronal population. At this time, most of the labeled cells are situated in the subventricular zone, although some immunoreactive Purkinje cells have already reached the cortex. Between embryonic day 17 and birth, four clusters of immunoreactive Purkinje cells appear in each hemicerebellum. Their time course and their pathways of migration to the cortex were followed. The immunoreactive clusters are tailed by a fibre-like immunostained material. The pattern of the migrating clusters at embryonic day 19 is very similar to the pattern of the corticonuclear projection observed at birth. From comparison between sections of embryos processed either for immunocytochemistry or Cresyl Violet staining, it appears that all the Purkinje cells are not immunoreactive. Positive and negative clusters of Purkinje cells are sharply delineated, their cells never mix. Immunopositive and negative clusters of Purkinje cells coexist until postnatal day 3. However, from birth onwards, negative clusters begin progressively in a caudorostral sequence to express guanosine 3':5'-phosphate-dependent protein kinase and rapidly attain the same level of immunoreactivity as previously labeled clusters. From postnatal day 5 all the Purkinje cells are immunoreactive. It is concluded that a compartmentalization of the cerebellar cortex is present very early and is evidenced by differences in the biochemical maturation of Purkinje cells. The axons of Purkinje cells reach the deep nuclei, following the same pathways as the clusters of Purkinje cells migrating to the cortex. Therefore, the mechanisms regulating the selection of the migratory routes followed by each Purkinje cell cluster are essential for the achievement of the topography of the corticonuclear projection. The level of protein kinase immunoreactivity cannot be taken as an index of the overall maturation of Purkinje cells, because it does not always coincide with the expression of other makers of biochemical and morphological differentiation of these neurons. During the early establishment of the cerebellar maps, an asynchrony in the expression of parts of the same genotype in the Purkinje cells may help in the establishment of ordered connections.     

Morphology and innervation of the teleost physostome swim bladders and their functional evolution in non-teleostean lineages

Swim bladders and lungs are homologous structures. Phylogenetically ancient actinopterygian fish such as Cladistians (Polypteriformes), Ginglymods (Lepisosteids) and lungfish have primitive lungs that have evolved in the Paleozoic freshwater earliest gnathostomes as an adaptation to hypoxic stress. Here we investigated the structure and the role of autonomic nerves in the physostome swim bladder of the cyprinid goldfish (Carassius auratus) and the respiratory bladder of lepisosteids: the longnose gar and the spotted gar (Lepisosteus osseus and L. oculatus) to demonstrate that these organs have different innervation patterns that are responsible for controlling different functional aspects. The goldfish swim bladder is a richly innervated organ mainly controlled by cholinergic and adrenergic innervation also involving the presence of non-adrenergic non-cholinergic (NANC) neurotransmitters (nNOS, VIP, 5-HT and SP), suggesting a simple model for the regulation of the swim bladder system. The pattern of the autonomic innervation of the trabecular muscle of the Lepisosteus respiratory bladder is basically similar to that of the tetrapod lung with overlapping of both muscle architecture and control nerve patterns. These autonomic control elements do not exist in the bladders of the two species studied since they have very different physiological roles. The ontogenetic origin of the pulmonoid swim bladder (PSB) of garfishes may help understand how the expression of these autonomic control substances in the trabecular muscle is regulated including their interaction with the corpuscular cells in the respiratory epithelium of this bimodal air-breathing fish.     

Localization of neurotransmitters, peptides and nNOS in the pseudobranchial neurosecretory cell system and associated carotid labyrinth of the catfish, Clarias batrachus

The carotid labyrinth is an enigmatic endocrine structure of unknown chemosensory function lying in the gill region of the catfishes. The carotid body is found at the carotid bifurcation of amphibians and all mammalian vertebrates on the evolutionary tree. It is a vascular expansion comprised of a cluster of glomus cells with associated (afferent and efferent) innervations. In the catfish species studied (Clarias batrachus) a neurosecretory cell system consisting of pseudobranchial neurosecretory cells connect the carotid labyrinth or large vessels (both the efferent branchial artery and dorsal aorta), and is likely akin to the glomus cells, but comparing these structures in widely divergent vertebrate species, the conclusion is that the structural components are more elaborate than those of terrestrial vertebrates. However, these cells reveal both an endocrine phenotype (such as the association with capillaries and large vessels) and the presence of regulatory substances such as neurotransmitters and neuropeptides producing good evidence for high levels of conservation of these substances that are present in the glomus cells of mammalian vertebrates. VIP-immunopositive neuronal cell bodies are detected in the periphery of the carotid labyrinth. They are presumptive local neurons that differ from pseudobranchial neurosecretory cells, the latter failing to express VIP in their soma.     

A lectin horseradish peroxidase study of the origin of ascending fibers in the medial forebrain bundle of the rat. The lower brainstem

The origins of projections within the medial forebrain bundle from the lower brainstem were examined with the horseradish peroxidase technique. Labeled cells were found in at least 15 lower brainstem nuclei following injections of a conjugate or horseradish peroxidase and wheat germ agglutinin at various levels of the medial forebrain bundle. Dense labeling was observed in the following cell groups (from caudal to rostral): A1 (above the lateral reticular nucleus); A2 (mainly within the nucleus of the solitary tract); a distinct group of cell trailing ventrolaterally from the medial longitudinal fasciculus at the level of the rostral pole of the inferior olive; raphe magnus; nucleus incertus; dorsolateral tegmental nucleus (of Castaldi); locus coeruleus; nucleus subcoeruleus; caudal part of the dorsal (lateral) parabrachial nucleus; and raphe pontis. Distinct but light labeling was seen in raphe pallidus and obscurus, nucleus prepositus hypoglossi, nucleus gigantocellularis pars ventralis, and the ventral (medial) parabrachial nucleus. Sparse labeling was observed throughout the medullary and caudal pontine reticular formation. Several lower brainstem nuclei were found to send strong projections along the medial forebrain bundle to very anterior levels of the forebrain. They were: A1, A2, raphe magnus (rostral part), nucleus incertus, dorsolateral tegmental nucleus, raphe pontis and locus coeruleus. With the exception of the locus coeruleus, attention has only recently been directed to the ascending projections of most of the nuclei mentioned above. Evidence was reviewed indicating that fibers from lower brainstem nuclei with ascending medial forebrain bundle projections distribute to widespread regions of the forebrain.It is concluded from the present findings that several medullary cell groups are capable of exerting a direct effect on the forebrain and that the medial forebrain bundle is the major ascending link between the lower brainstem and the forebrain.     

Differential expression of NMDA receptors in serotonergic and/or GABAergic neurons in the midbrain periaqueductal gray of the mouse

N-methyl-d-aspartate (NMDA) receptors expressed in the midbrain periaqueductal gray (PAG) exert various physiological functions. The PAG contains various neurotransmitter phenotypes, which include GABAergic neurons and serotonergic neurons. In the present experiments, we made tight-seal whole-cell recordings from GABAergic and/or serotonergic neurons in mouse PAG slices and analyzed NMDA and non-NMDA receptor-mediated excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation. The NMDA/non-NMDA ratio of EPSC amplitude was high and the decay time course of NMDA-EPSC was slow in non-serotonergic/GABAergic neurons. In contrast, serotonergic neurons exhibited a low NMDA/non-NMDA ratio and a fast decay time course of NMDA-EPSC. Peripheral nerve ligation-induced chronic pain was associated with an increased NMDA/non-NMDA ratio in serotonergic neurons. Additionally, single-cell real-time RT-PCR analysis showed that peripheral nerve ligation up-regulated NR2B subunit expression in non-serotonergic/non-GABAergic neurons. Such changes in NMDA receptor expression in the PAG result in an alteration of the descending modulation of nociception, which might be an underlying mechanism for peripheral nerve injury-evoked persistent pain. Finally, the expression of NMDA receptors seems differentially regulated among neurons of different neurotransmitter phenotypes in the PAG.     

Immunolocalization of neurotransmitter-synthesizing enzymes and neuropeptides with associated receptors in the photophores of the hatchetfish, Argyropelecus hemigymnus Cocco, 1829

Anatomical and functional studies of the autonomic innervation of the photophores of luminescent fishes are scarce. The present immunohistochemical study demonstrated the presence of nerve fibers in the luminous epithelium and lens epithelium of the photophores of the hatchet fish, Argyropelecus hemigymnus and identified the immunoreactive elements of this innervation. Phenylethanolanine N-methyltransferase (PNMT) and catecholamine (CA)-synthesizing enzymes were detected in nerve varicosities inside the two epithelia. Neuropeptides were localized in neuropeptide Y (NPY) and substance P (SP)- and its NK11 receptor-immunopositive nerves in the lens epithelium. Neuropeptides were also localized in non-neural cell types such as the lens cells, which displayed immunoreactivities for pituitary adenylate cyclase activating peptide (PACAP) and their receptors R-12 and 93093-3. This reflects the ability of the neuropeptide-containing nerves and lens cells to turn on and off the expression of selected messengers. It appears that the neuropeptide-containing nerves demonstrated in this study may be sensory. Furthermore, neuronal nitric oxide synthase-immunopositive axons associated with photocytes in the luminous epithelium have previously been described in this species. Whereas it is clear that the photophores receive efferent (motor) fibers of spinal sympathetic origin, the origin of the neuropeptide sensory innervation remains to be determined. The functional roles of the above neuropeptides or their effects on the bioluminescence or the chemical nature of the terminals, either sensory or postganglionic neurons innervating the photophores, are still not known.     

Specificity of peripheral nerve regeneration: interactions at the axon level

Peripheral nerves injuries result in paralysis, anesthesia and lack of autonomic control of the affected body areas. After injury, axons distal to the lesion are disconnected from the neuronal body and degenerate, leading to denervation of the peripheral organs. Wallerian degeneration creates a microenvironment distal to the injury site that supports axonal regrowth, while the neuron body changes in phenotype to promote axonal regeneration. The significance of axonal regeneration is to replace the degenerated distal nerve segment, and achieve reinnervation of target organs and restitution of their functions. However, axonal regeneration does not always allows for adequate functional recovery, so that after a peripheral nerve injury, patients do not recover normal motor control and fine sensibility. The lack of specificity of nerve regeneration, in terms of motor and sensory axons regrowth, pathfinding and target reinnervation, is one the main shortcomings for recovery. Key factors for successful axonal regeneration include the intrinsic changes that neurons suffer to switch their transmitter state to a pro-regenerative state and the environment that the axons find distal to the lesion site. The molecular mechanisms implicated in axonal regeneration and pathfinding after injury are complex, and take into account the cross-talk between axons and glial cells, neurotrophic factors, extracellular matrix molecules and their receptors. The aim of this review is to look at those interactions, trying to understand if some of these molecular factors are specific for motor and sensory neuron growth, and provide the basic knowledge for potential strategies to enhance and guide axonal regeneration and reinnervation of adequate target organs.