Sensory pathways in amphioxus larvae I. Constituent fibres of the rostral and anterodorsal nerves, their targets and evolutionary significance

Serial and interval electron micrograph series were used to examine the rostral and anterodorsal nerves of 12.5‐day‐old amphioxus larvae and trace selected fibres to their targets in the nerve cord. The nerves contain a variety of fibre types, including axons from at least two types of epithelial sensory cells and neurites derived from dorsal (Retzius) bipolar cells located within the cord. The rostral epithelial cells form basal synapses with a population of peripheral neurites that probably derive from the dorsal bipolar cells, though other sources are possible. Varicosities containing dense‐core vesicles occur at the tip of the rostrum, indicating the presence of efferent innervation at this site. Within the cord, some peripherally derived rostral afferents terminate at the level of the anterior cerebral vesicle, others synapse directly with both motoneurones and the notochord, but those in the largest bundle target the dendrites of the large paired neurones (LPNs) located in the primary motor centre. LPN dendrites also receive synapses from sensory fibres arriving via the anterodorsal nerves, from the anterior‐most of the dorsal bipolar cells, referred to here as tectal cells, and from a single fibre derived from the frontal eye. This convergence of multiple inputs accords with other evidence that the LPNs are key intermediaries in the sensorimotor pathway that activates the larval escape response. The rostral nerves are much larger at metamorphosis, but the ventral tracts that derive from them are still comparatively small. This is because the majority of rostral fibres are diverted into a late‐developing dorsal tract that travels within the cord to the front end of the dorsolateral neuropile, where most of its fibres disperse and form synapses. The positioning of the dorsal and ventral tracts strongly suggests homology with vertebrate olfactory and terminal nerves, respectively. This, and the question of whether the amphioxus central nervous system has anything comparable to the olfactory bulb, a telencephalic structure, is discussed.     

Distribution Pattern of γ-amino Butyric Acid Immunoreactive Neural Structures in the Central and Peripheral Nervous System of the Tubificid Worm, Limnodrilus hoffmeisteri

By means of whole-mount immunohistochemistry, putative inhibitory (GABAergic) neural structures were identified in the central and peripheral nervous system of the tubificid worm, Limnodrilus hoffmeisteri. In the supraoesophageal ganglion (brain) only few strongly labelled cells were observed. However, in its commissural part a high number of stained nerve fibres, arising mainly from the ventral nerve cord and prostomium, occurred. Except for the suboesophageal ganglion the arrangement of γ-amino butyric acid-immunoreactive (GABA-IR) structures proved to be identical in each VNC ganglion. Behind the first segmental nerves three pairs of heavily stained neurones were located. Their processes (both ipsi- and contralateral) form four bundles of fine-fibred polysegmental interneuronal tracts that run close to the dorsal giant axons from the terminal ganglion to the suboesophageal one without interruption. A few small motoneurons and a pair of large ones with contralateral processes were also identified. A bipolar (presumably sensory) neuron was located at the root of each second segmental nerve. GABA-IR neurons were also found in the stomatogastric ganglia and pharyngeal wall; however, the latter structure had a well-developed fibre network, as well. Present results suggest that GABA acts as a common neurotransmitter in sensory, interneuronal and motor system of L. hoffmeisteri. The possible functional role of the identified GABA-IR neural structures in locomotion, escape and withdrawal reflexes in tubificid worms is discussed.     

The central nervous system, its cellular organisation and development, in the tadpole larva of the ascidian Ciona intestinalis

From its numerical composition, the central nervous system (CNS) of the ascidian larva is one of the simplest known nervous systems having a chordate plan. Fewer than 350 cells together constitute a caudal nerve cord, an interposed visceral ganglion containing motor circuits for swimming and, rostrally, an expanded sensory vesicle containing major sensory and interneuron regions of the CNS. Some cells are ependymal, with ciliated surfaces lining the neural canal, while others are clearly either sensory receptors or motoneurons, but most are distinguishable only on cytological grounds. Although reassignments between categories are still being made, there is evidence for determinancy of total cell number. We have made three-dimensional cell maps either from serial semithin sections, or from confocal image stacks of whole-mounted embryos and larvae stained with nuclear markers. Comparisons between the maps of neural tubes in embryos of successive ages, that is, between cells in one map and their progeny in older maps, enable us to follow the line of mitotic descent through successive maps, at least for the caudal neural tube. Details are clear for the lateral cell rows in the neural tube, at least until the latter contains approximately 320 cells, and somewhat for the dorsal cell row, but the ventral row is more complex. In the hatched larva, serial-EM reconstructions of the visceral ganglion reveal two ventrolateral fibre bundles at the caudalmost end, each of 10-12 axons. These tracts include at least five pairs of presumed motor axons running into the caudal nerve cord. Two pairs of axons decussate. Complementing this vertebrate feature in the CNS of the larval form of Ciona, we confirm that synapses form upon the somata and dendrites of its neurons, and that its motor tracts are ventral.     

K252a Modulates the Expression of Nerve Growth Factor-Dependent Capsaicin Sensitivity and Substance P Levels in Cultured Adult Rat Dorsal Root Ganglion Neurones

Abstract: K252a, an inhibitor of trk phosphorylation and nerve growth factor signal transduction in PC12 cells, blocked nerve growth factor-induced responses in cultured adult rat dorsal root ganglion sensory neurones. The nerve growth factor-dependent appearance of capsaicin sensitivity and accumulation of the neuropeptide substance P were inhibited when dorsal root ganglion neurones were grown in the presence of low concentrations (100 n M ) of K252a. At higher concentrations (3 µ M ), however, K252a stimulated the development of capsaicin sensitivity and the accumulation of substance P even in the absence of nerve growth factor. By using a wide dose range, therefore, we showed that K252a could either inhibit or mimic nerve growth factor's actions on sensory neurones. These results may explain the apparent paradox in the literature that some groups show a blocking effect of K252a on nerve growth factor-dependent survival of dorsal root ganglion sensory neurones, whereas others report that K252a can substitute for nerve growth factor or other trophic factors and promote neuronal survival.     

Epidermal receptor development and sensory pathways in vitally stained amphioxus (Branchiostoma floridae)

Chloromethyl (CM) DiI was applied to the exterior of living embryos, larvae, and metamorphic juveniles of amphioxus. This fluorescent dye is taken up preferentially (but not highly selectively) by epidermal receptors and often stains sensory axons to their full extent. Type I primary receptors in the epidermis first become morphologically detectable along the rostrocaudal axis of the 2.5 day larva when their epidermal perikarya extend unbranched axons to the nerve cord. These axons run posteriorly or anteriorly within the nerve cord, depending on whether their perikarya are located, respectively, rostral or caudal to the most posterior pharyngeal slit. In later larvae, axons of type I receptors are organized into a dorsal and a subdorsal sensory tract on either side of the nerve cord. In the epidermis of metamorphic juveniles, CM-DiI also stains type II receptors (which are axonless, secondary receptors) and ventral pit cells (which may not be receptors). It is probable, but not yet conclusively demonstrated, that peripheral neurites from Retzius bipolar cells (primary intramedullary sensory neurones) synapse with type II secondary epidermal receptors or ramify freely among the other epidermal cells. The discussion considers homologies among epidermal sensory receptors in chordates.     

Primary and secondary somatosensory projections in direct-developing Plethodontid Salamanders

In three species of plethodontid salamanders (Plethodon jordani, Hydromantes italicus, and Bolitoglossa subpalmata), primary and secondary somatosensory pathways were investigated by means of tract-tracing in vivo and in vitro using biocytin, horseradish peroxidase, and neurobiotin. Afferent sensory fibers of cranial nerves V, VII, and X and the brachial nerve run in the dorsal funiculus of the medulla oblongata and spinal cord. Fibers ascend to the level of, but do not enter, the cerebellum. In the caudal medulla oblongata, sensory tracts of the cranial nerves descend in a dorsal and a dorsolateral bundle and reach the level of the fourth spinal nerve. Two bundles are likewise formed by spinal afferent fibers, which descend to the level of the seventh spinal nerve. Secondary somatosensory projections ascend in contralateral ventral, contralateral lateral, and ipsilateral lateral tracts, the latter two corresponding to the spinal lemniscal tracts of Herrick. These tracts reach the cerebellum, mesencephalic, and diencephalic targets (tegmentum, torus, tectum, tuberculum posterius, pretectum, and ventral thalamus) ipsi- and contra-laterally. The projection to the tectum is confined to fiber layer 4. Fibers of the ascending tracts cross in the cerebellar and tectal commissure. Our study demonstrates that the ascending secondary somatosensory pathways of plethodontid salamanders differ remarkably from those of other amphibians. J. Morphol. 238:307–326, 1998. © 1998 Wiley-Liss, Inc.     

Ventral neurons in the anterior nerve cord of amphioxus larvae. I. An inventory of cell types and synaptic patterns

Serial sections were used to map the ventrally positioned neurons of the anterior nerve cord of a 12.5-day amphioxus larva from the infundibular region to the end of somite 2. Synaptic patterns reveal five categories of descending pathways, four of which are associated with the ventral compartment (VC) motoneurons responsible for escape swimming. 1) Pre-, para-, and postinfundibular (tegmental) neurons with large varicosities and mixed vesicle populations provide both synaptic and paracrine input to various components of the tegmental neuropile and primary motor center. Four categories of these neurons are distinguished on the basis of their vesicles. 2) Multiple anterior sensory pathways converge on the large paired neurons (LPNs) located near the junction of somites 1 and 2. LPN synaptic output is almost exclusively contralateral. This, together with the evidence for cross-innervation between the third pair of LPNs, is consistent with the latter acting as locomotory pacemakers. 3) Axons from several classes of tegmental neurons converge in the paraxial region on each side of the cord where they form distinct tracts, the upper paraxial bundles. The right bundle is larger than the left, which suggests a role during early development when myotome contractions are biased to one side. 4) Fibers in the ventral tracts from ipsilateral projection neurons, sensory neurons, and additional ascending fibers synapse repeatedly with VC motoneurons. This may be how the overall level of excitation of the latter is controlled so as to modulate their response to pacemaker input. The fifth pathway consists of fibers involved in controlling the dorsal compartment (DC) motoneurons responsible for slow swimming, which are largely isolated from inputs to the VC locomotory system. The ventral neurons of the primary motor center form a more or less continuous file on either side of the floor plate, with certain cell types showing a tendency to cluster. There are, however, few obvious patterns of the kind expected if development were controlled by a rigid, lineage-based mechanism. The evolutionary implications of the involvement of a midbrain-level pacemaker in controlling larval swimming in amphioxus is discussed.     

Localization of nerve growth factor-like immunoreactivity in rat nervous tissue

The use of immunofluorescence with affinity-purified antibodies enabled cytological localization of nerve growth factor-like material in the rat. Immunoreactivity was observed along various nerve tracts of the foetal rat brain and spinal cord at day 15 of gestation. Longitudinal pathways in ventral and dorsal spinal cord, ventral lower brain stem, posterior commissure, retroflex fascicle and in the olfactory bulb were all positive. A weaker and more widely spread immunostaining was visible in many areas in the central nervous system. Cranial nerves were strongly immunoreactive. Neuronal perikarya in the retina and the olfactory mucosa as well as filae olfactoriae and the olfactory nerve all the way to the olfactory bulb were also positive. In sensory ganglia and peripheral nerves most immunoreactivity was confined to supporting tissues, probably including Schwann cells. In irides, the pattern of immunoreactivity was similar to that of the sensory and autonomic innervation. More intensively fluorescent material was found in regrowing nerve fibres in iris transplants. Our histochemical results suggest that nerve growth factor and/or a related protein is present in large amounts along nerve pathways in supportive tissues of the peripheral nervous system as well as in the central nervous system during early development.     

Prolonged primary afferent induced alterations in dorsal horn neurones, an intracellular analysis in vivo and in vitro

1.) Peripheral tissues injury produces long lasting sensory and motor disturbances in man that present as the post-injury hypersensitivity syndrome with a reduction in the threshold required to elicit either pain or the flexion withdrawal reflex and an exaggeration of the normal response to suprathreshold stimuli. 2.) Two mechanisms contribute to these changes; sensitization of the peripheral terminals of high threshold primary afferents and an increase in the excitability of the spinal cord; a phenomenon known as central sensitization. 3.) Central sensitization has previously been shown by our laboratory to be the consequence of activity in unmyelinated primary afferents. Brief (20 s) C-fibre strength conditioning stimuli have the capacity to produce both a prolonged heterosynaptic facilitation of the flexion reflex and an alteration in the response properties of dorsal horn neurones, that long outlast the conditioning stimulus. 4.) In the adult decerebrate-spinal rat preparation we have, using intracellular recordings of dorsal horn neurones, examined the time course of the central effects of different types of orthodromic inputs. The hemisected spinal cord preparation isolated from 12-14 day rat pups has been used to see whether prolonged alterations in dorsal horn properties induced by orthodromic inputs can be studied in vitro. 5.) Single stimuli applied to a cutaneous nerve at graded strengths to successively recruit A beta, A delta and C-afferents produce, in the majority of neurones recorded in the deep dorsal horn in vivo, a series of post synaptic potentials that last from between ten and several hundred milliseconds. 6.) Repeated low frequency stimulation of C but not A-afferent fibres results in a pattern of progressive response increment or windup in a proportion of dorsal horn neurones. In some of the neurones the windup is associated with a depolarization that outlasts the stimulus period for tens of seconds. 7.) Application of the chemical irritant mustard oil to the skin activates chemosensitive C-afferent fibres for 1-3 minutes. Such a conditioning stimulus results however in an expansion in the size and an alteration in the response properties of the receptive fields of dorsal horn neurones that lasts for tens of minutes. 8.) In dorsal horn neurones recorded intracellularly in the isolated hemisected spinal cord, both intrinsic membrane properties and the orthodromic responses to primary afferent input can be studied. Repeated stimulation of a dorsal root produces in some neurones a prolonged heterosynaptic facilitation with both an augmentation of the response to the conditioning root (homosynaptic potentiation) and to adjacent test roots (heterosynaptic potentiation).(ABSTRACT TRUNCATED AT 400 WORDS)     

The structure of the ventral nerve cord of Caenorhabditis elegans.

The nervous system of Caenorhabditis elegans is arranged as a series of fibre bundles which run along internal hypodermal ridges. Most of the sensory integration takes place in a ring of nerve fibres which is wrapped round the pharynx in the head. The body muscles in the head are innervated by motor neurones in this nerve ring while those in the lower part of the body are innervated by a set of motor neurones in a longitudinal fibre bundle which joins the nerve ring, the ventral cord. These motor neurones can be put into five classes on the basis of their morphology and synaptic input. At any one point along the cord only one member from each class has neuromuscular junctions. Members of a given class are arranged in a regular linear sequence in the cord and have non-overlapping fields of motor synaptic activity, the transition between fields of adjacent neurones being sharp and well defined. Members of a given class form gap junctions with neighbouring members of the same class but never to motor neurones of another class. Three of the motor neurone classes receive their synaptic input from a set of interneurones coming from the nerve ring. These interneurones can in turn be grouped into four classes and each of three motor neurone classes receives its synaptic input from a unique combination of interneurone classes. The possible developmental and functional significance of these observations is discussed.     

The central projection of cephalic mechanosensory axons in the haematophagous bug Triatoma infestans

The projections of mechanosensory hairs located on the dorsal and lateral head of the adult haematophagous bug Triatoma infestans were analyzed by means of cobalt filling. Axons run into the anterior and posterior tegumentary nerve and project through the brain to the ventral nerve cord. The fibres are small in diameter and run as a fascicle. Some branches run into suboesophageal and prothoracic centres; others run as far as to the mesothoracic ganglion. These sensory projections resemble that of wind-sensitive head hairs of the locust. The functional role of this sensory system in this species is discussed.     

低温保存许旺细胞对周围神经再生的作用

目的：比较原代培养许旺细胞（Schwann cells,SCs)和冷冻保存的SCs移植对损伤后坐骨神经再生的作用。方法：原代培养和液氮保存的SCs分别移植到桥接缺损坐骨神经的硅胶管内。在移植后不同时间（第6和8周末）,硅胶管远端神经干内注射HRP,逆行追踪背根神经节和脊髓前角的标记神经元数量;测量再生神经纤维的复合动作电位传导速度;电镜观察再生神经纤维的髓鞘形成。结果：原代培养和冷冻保存SCs在移植后不同时间其背根神经节和脊髓前角神经元HRP标记细胞数量、再生神经纤维的复合动作电位传导速度基本一致,再生神经纤维髓鞘的形成未见明显差别。结论：冷冻保存的SCs仍具有促进损伤后周围神经再生的能力。     

Electrophysiological responses from the peripheral olfactory system of the American eel,Anguilla rostrata

Summary A method is described for recording with microelectrodes from central neurones in locusts,Schistocerca gregaria americana, that are free to perform a large fraction of their behavioural repertoire. This tethered preparation has been used to examine the individual responses of large neurones in the neck connectives to a range of sensory stimuli.From differences in the responses of the units examined and from their positions in the connective, as determined by dye iontophoresis, 31 separate neurones have been identified. The axons of these cells had relatively constant diameters and cord positions in different animals and appeared in both right and left connectives but with their positions mirror reversed. The majority of these 31 cells carried descending information from the head ganglia and under our experimental conditions, 7 were found to have wind stimulation as their strongest sensory input, 17 had visual stimulation, 4 had sound stimulation and 3 had proprioceptive input.Abbreviations DCMD descending contralateral movement detector (neurone) - DIMD descending ipsilateral movement detector (neurone)     

Changes in NADPH-diaphorase activity in the rat dorsal horn following an acute experimental myositis

Previous neurophysiological experiments have shown that in rats with an acute myositis of the gastrocnemius-soleus muscle, dorsal horn neurones exhibit an increase in responsiveness to peripheral stimulation and in background activity. The present study investigated the possible correlation between changes in NADPH-diaphorase activity and neurophysiological alterations. In the animals used for the electrophysiological experiments the diaphorase activity in sections of the lumbar spinal cord was determined with the NADPH-nitroblue tetrazolium reaction. The main finding was a massive reduction in the number of diaphorase-positive cells in the superficial dorsal horn in animals with a myositis. The staining intensity in the remaining neurones was unchanged. The results are interpreted as indicating that the myositis in addition to the surgical operations represents a supramaximal input to the dorsal horn causing neurotoxic effects in diaphorase- positive neurones.     

PKCγ免疫反应定位小鼠皮质脊髓束的实验研究

目的探讨显示皮质脊髓束在成年小鼠脑和脊髓中定位分布的简便有效方法。方法运用蛋白激酶Cγ（PKCγ）免疫组织化学染色法,观察成年ICR小鼠脑和脊髓中皮质脊髓束的定位和分布情况。结果PKCγ免疫阳性产物分布于大脑运动皮层第V层锥体细胞胞体和轴突中,锥体细胞的阳性纤维经内囊、中脑大脑脚底、脑桥基底部、下行至延髓锥体中。在延髓下段,PKCγ阳性纤维经锥体交叉后进入对侧脊髓灰质后联合背侧,形成背侧皮质脊髓束,在脊髓白质的后索腹侧深层下行,至骶髓3-4节段以下逐渐消失。在整个脊髓前索和外侧索中未见有PKCγ阳性纤维。结论PKCγ特异地表达于脊髓后索皮质脊髓束中,提示PKCγ免疫组织化学法是一种显示和观察皮质脊髓束精确定位的有效方法。     

Distribution of binding sites for the plant lectin Ulex europaeus agglutinin I on primary sensory neurones in seven different mammalian species

There is an increasing body of evidence to suggest that different functional classes of neurones express characteristic cell-surface carbohydrates. Previous studies have shown that the plant lectin Ulex europaeus agglutinin-I (UEA) binds to a population of small to medium diameter primary sensory neurones in rabbits and humans. This suggests that a fucose-containing glycoconjugate may be expressed by nociceptive primary sensory neurones. In order to determine the extent to which this glycoconjugate is expressed by other species, in the current study, we have examined the distribution of UEA-binding sites on primary sensory neurones in seven different mammals. Binding sites for UEA were associated with the plasma membrane and cytoplasmic granules of small to medium dorsal root ganglion cells and their axon terminals in laminae I–III of the grey matter of the spinal cord, in the rabbit, cat and marmoset monkey. However, no binding was observed in either the dorsal root ganglia or spinal cord in the mouse, rat, guinea pig or flying fox. These results indicate an inter-species variation in the expression of cell-surface glycoconjugates on mammalian primary sensory neurones.     

Effects of Q-switched Nd:YAG laser irradiation on neural impulse propagation: I. Spinal cord.

We studied the effect of irradiation with Q-switched Nd:YAG laser light (1064 nm) on spinal cord dorsal column and dorsolateral white matter in anesthetized rats. To evoke a synchronous sensory input, the sciatic nerve was stimulated electrically and the resulting evoked spinal cord potential (SCP) recorded from the dorsal columns of the ipsilateral side. The waveshape of the SCP showed three components: an early positive peak (P1), representing the responses of the most rapidly conducting fibers, followed by two negative peaks (N1 and N2), which are mainly due to synaptic effects of the volley on dorsal horn cells located in dorsal grey matter. Laser irradiation at 50 mJ/pulse and above resulted in severe reduction in the amplitudes of N1 and N2. In contrast, there was either no reduction at all or only a slight decrease in the amplitude of P1. The selective loss of the synaptic field might be mediated by impairment of synaptic transmission or by loss of high threshold fiber input to dorsal horn neurones. In either event it is likely that the mechanism of the differential effects of laser irradiation on the components of the electrically evoked SCP is at least in part photothermally mediated, since intracord temperatures during laser application greatly exceeded the physiological range.     

Nerve growth factor-induced stimulation of dorsal root ganglion/spinal cord co-grafts in oculo: enhanced survival and growth of CGRP-immunoreactive sensory neurons

Intraocular co-grafts of rat fetal spinal cord and dorsal root ganglia were used to examine the enhanced survival, growth, and differentiation of sensory neurons by nerve growth factor. E14 lumbar spinal segments were implanted into the anterior eye chamber of capsaicin-pretreated rats. Two weeks later, an E14 dorsal root ganglion was implanted beside the spinal cord graft. Nerve growth factor or vehicle was injected weekly for 4 weeks into the anterior eye chamber. Co-grafts were examined weekly and, at 6 weeks, processed for calcitonin gene-related peptide (CGRP) immunofluorescence. No differences in overall size were determined for the grafts. Co-grafts treated with nerve growth factor contained many more CGRP neurons (19.4 cells/20 microm) that were significantly larger (mean 764 microm2) than neurons from control co-grafts (8.6 cells/20 microm; mean 373 microm2). In co-grafts treated with nerve growth factor, CGRP-immunoreactive fibers were extensive in the dorsal root ganglion, adjacent iris, and spinal cord compared to control co-grafts. A few CGRP-positive motoneurons were observed in the spinal cord, but no differences in number or size of motoneurons were found. The current report demonstrates that spinal cord and dorsal root ganglia can be co-grafted in oculo for long periods of time. Many dorsal root ganglion neurons survive and send peripheral processes into the iris and central processes into the spinal cord under the influence of exogenous nerve growth factor. The intraocular graft paradigm can be of use to further examine the role of neurotrophic factors in regulating or modulating dorsal root ganglion and spinal cord neurons.     

Antennal movements induced by odour and central projection of the antennal neurones in the honey-bee

Movements of the antennae induced by odour were investigated. Odour presented to the antenna of one side induced both antennae to move to that side. The EMGs recorded from the flexor muscles of both scapes showed that the latency of the movement of the ipsilateral flagellum when induced by odour was about 71 msec shorter than that of the contralateral flagellum. Recording electrical activities from the antennal nerve showed that there are more than 14 neurones in the antenno-motor externus.The distribution of the antennal nerve in the brain was investigated histologically by the injection of fluorescent dye. Antennal sensory neurones terminated at the glomeruli in the antennal lobe, in the dorsal lobe, in the protocerebrum, and in the commissural part of the suboesophageal ganglion. Injection of the fluorescent dye into the antennal nerve after degeneration of the antennal sensory neurones showed that the antennal motoneurones run in the ventral side of the antennal and dorsal lobes, and terminate in the marginal region of the ipsilateral oesophageal connective.The difference in latency of odour-induced flagellar movements is discussed in relation to the histological results and the unitary responses in the brain reported previously.