Fine structure of normal and degenerating primary afferent boutons associated with characterized spinocervical tract neurons in the cat

Spinocervical tract neurons in the dorsal horn of the cat spinal cord were intracellularly stained with horseradish peroxidase. The neurons came from one intact animal and from animals with dorsal rhizotomies (L3-S2) 3, 5, 10, 28 and 42 days previously. The morphology of terminals associated with spinocervical tract neurons was examined in a combined light and electron microscopical study. Some terminals containing agranular, circular vesicles degenerated as a result of deafferentation; these are therefore the terminals forming monosynaptic inputs to the neurons from primary afferent fibres. Other terminals containing agranular circular vesicles and terminals containing ovoid agranular vesicles survived deafferentation; these boutons therefore do not originate from primary afferent fibres.     

Calcitonin gene-related peptide-positive neurons and fibers in the cat dorsal column nuclei

An antiserum raised against the C-terminal region of rat alpha-calcitonin gene-related peptide has been used to investigate the morphology and topographical distribution of neurons and terminals containing calcitonin gene-related peptide in the cat dorsal column nuclear complex. Calcitonin gene-related peptide-positive fibers and axon terminals were denser in the cuneate nucleus than in the other dorsal column nuclei subdivisions and were observed throughout all rostrocaudal levels. They were densest in the dorsal and ventrolateral portions of the middle cuneate. Immunoreactive neurons were observed only in animals pre-treated with colchicine. In these cases, some calcitonin gene-related peptide-positive neurons were present in the cuneate and in the external cuneate. In double-labeling experiments, visualization of calcitonin gene-related peptide immunoreactivity in dorsal root ganglia neurons was combined with the retrograde transport of colloidal gold-labeled wheat germ agglutinin conjugated to inactive horseradish peroxidase injected in the cuneate nucleus. These experiments show that calcitonin gene-related peptide-positive fibers in the cuneate nucleus originate mostly from C3-C6 medium sized dorsal root ganglia neurons but also from some small and large neurons. These results suggest that calcitonin gene-related peptide-positive fibers may convey sensory information from a wide range of peripheral receptors.     

Immunocytochemical evidence for a serotoninergic innervation of dorsal column postsynaptic neurons in cat and monkey: Light- and electron-microscopic observations

Dorsal column postsynaptic neurons in the lumbosacral enlargements of cats and a monkey were retrogradely labeled by placing horseradish peroxidase on their severed axons in the thoracic dorsal columns. After visualizing the retrogradely-labeled neurons, the tissue was immunocytochemically stained with an antiserum directed against serotonin. Immunoreactive axonal varicosities contacted the perikarya and proximal dendrites of every retrogradely-labeled neuron examined in cat (mean 61 contacts/cell) and nearly every neuron in the monkey (mean 18 contacts/cell). Electron microscopy showed that the immunoreactive axonal varicosities contained pleomorphic (round to oval) agranular vesicles and formed symmetrical synapses on retrogradely-labeled neurons.

It is concluded that dorsal column postsynaptic neurons are innervated directly by the brain stem's descending, serotoninergic system(s).     

内脏伤害性刺激在脊髓突触后背柱神经元的信号转导机制

内脏痛是急慢性胃肠道、盆腔、泌尿道和其它实体器官疾病的最常见症状之一。许多疾病伴发的内脏性疼痛十分顽固,如肠道激惹综合征、间质性膀胱炎、胰腺炎、子宫内膜异位症和癌性内脏痛等,给临床医师带来了极大的挑战。内脏疼痛机制十分复杂。近年来,一系列的临床和基础研究发现起源于     

Postsynaptic dorsal column neurons in the cat: a study with retrograde transport of horseradish peroxidase

Summary In situ hybridization histochemistry and RNA blots were used to study the expression of glutamic acid decarboxylase (GAD) mRNA in rats with or without a unilateral lesion of midbrain dopamine neurons. Two populations of GAD mRNA positive neurons were found in the intact caudate-putamen, substantia nigra and fronto-parietal cortex. In caudate-putamen, only one out of ten of the GAD mRNA positive neurons expressed high levels, while in substantia nigra every second of the positive neurons expressed high levels of GAD mRNA. Relatively few, but intensively labelled neurons were found in the intact fronto-parietal cerebral cortex. In addition, one out of six of the GAD mRNA positive neurons in the fronto-parietal cortex showed a low labeling. On the ipsilateral side, the forebrain dopamine deafferentation induced an increase in the number of neurons expressing high levels of GAD mRNA in caudateputamen, and a decrease in fronto-parietal cortex. A smaller decrease was also seen in substantia nigra. However, the total number of GAD mRNA positive neurons were not significantly changed in any of these brain regions. The changes in the levels of GAD mRNA after the dopamine lesion were confirmed by RNA blot analysis. Hence, midbrain dopamine neurons appear to control neuronal expression of GAD mRNA by a tonic down-regulation in a fraction of GAD mRNA positive neurons in caudate-putamen, and a tonic up-regulation in a fraction of GAD mRNA positive neurons in fronto-parietal cortex and substantia nigra.     

Comparative study of viscerosomatic input onto postsynaptic dorsal column and spinothalamic tract neurons in the primate.

The purpose of the present investigation was to examine, in the primate, the role of the postsynaptic dorsal column (PSDC) system and that of the spinothalamic tract (STT) in viscerosensory processing by comparing the responses of neurons in these pathways to colorectal distension (CRD). Experiments were done on four anesthetized male monkeys (Macaca fascicularis). Extracellular recordings were made from a total of 100 neurons randomly located in the L(6)-S(1) segments of the spinal cord. Most of these neurons had cutaneous receptive fields in the perineal area, on the hind limbs or on the rump. Forty-eight percent were PSDC neurons activated antidromically from the upper cervical dorsal column or the nucleus gracilis, 17% were STT neurons activated antidromically from the thalamus, and 35% were unidentified. Twenty-one PSDC neurons, located mostly near the central canal, were excited by CRD and three were inhibited. Twenty-four PSDC neurons, mostly located in the nucleus proprius, did not respond to CRD. Of the 17 STT neurons, 7 neurons were excited by CRD, 4 neurons were inhibited, and 6 neurons did not respond to CRD. Of the unidentified neurons, 23 were excited by CRD, 7 were inhibited, and 5 did not respond. The average responses of STT and PSDC neurons excited by CRD were comparable in magnitude and duration. These results suggest that the major role of the PSDC pathway in viscerosensory processing may be due to a quantitative rather than a qualitative neuronal dominance over the STT.     

Interactions between spinocervicolemniscal and dorsal column pathways in thalamic ventrobasal neurons in the cat.

Ventrobasal (VB) thalamic cells responding to electrical stimulation of the dorsal column (DC) and of the dorsolateral funiculus DLF in cats with DC intact or cut at C4 and isolated, were studied. Among the cells observed, about 30% showed convergence to the two pathways. Different patterns of interaction between the two inputs were found, but the most frequent effect observed was an inhibition exerted by dorsal column (conditioning) stimulation on the activity evoked by dorsolateral funiculi (test) stimulation. From these results a dominance of the DC over the spinocervico-thalamic (SCT) inputs was confirmed.     

Possible neurotransmitters of the dorsal column afferents: effects of dorsal column transection in the cat

We investigated quantitative changes in markers of possible neurotransmitters in the dorsal column nuclei following transection of the dorsal column in the cat. Seven days after unilateral transection of the dorsal column at the upper cervical level, choline acetyltransferase activity and concentrations of glutamate, aspartate, gamma-aminobutyrate and substance P were measured throughout the longitudinal axis of the dorsal column nuclei. In addition, high-affinity uptake of choline, D-aspartate and gamma-aminobutyrate into the synaptosomal fraction of the dorsal column nuclei were also measured. Choline acetyltransferase activity and high-affinity choline uptake were reduced by approx. 30% on the caudal to the obex. Reduction of high-affinity uptake of D-aspartate by approx. 30% was observed on the operated side in the central part of these nuclei, although the decrease in glutamate and aspartate was not significant in the nuclei on the operated side compared with that on the intact side. No significant changes were found in the high-affinity uptake of gamma-aminobutyrate or the contents of gamma-aminobutyrate and substance P in any areas of the dorsal column nuclei. These results suggest that not only glutamate and/or aspartate but also acetylcholine may be neurotransmitter candidates for the ascending fibres terminating in the dorsal column nuclei, whereas there may be few fibres containing substance P or gamma-aminobutyrate in the dorsal column.     

Spinocervical neurons and dorsal horn neurons projecting to the dorsal column nuclei through the dorsolateral fascicle: a retrograde HRP study in the cat

Summary Spinocervical cells were identified by retrograde labelling from implants of HRP in the dorsolateral fascicle after destruction of the dorsal columns. They lay in laminae III and IV throughout the cord in estimated numbers of 700, 450 and 1100 in lumbosacral enlargement, upper lumbar and thoracic cord, and brachial enlargement respectively. In the cord enlargements dendritic trees were mainly or exclusively developed dorsally, with rostrocaudal exceeding mediolateral spread, and a gradient across the dorsal horn, lateral cells showing this contrast most strongly. Dendritic spread was limited at the II/III laminar boundary. Transition occurred at the edge of the enlargements to a shape with extreme rostrocaudal elongation of perikarya and of dendritic trees in upper lumbar and thoracic segments. Axons of Spinocervical cells ascended in the most dorsal part of the fascicle, distinguishable from the larger spinocerebellar bundle lying adjacent and ventral. The initial axonal course was tortuous, with local collateral branching, the axon sometimes travelling briefly in the dorsal column. In other experiments implants were made ipsilaterally in the dorsal column nuclei after destruction of the dorsal columns. Cells were few and relatively poorly labelled, for which the reasons are discussed. Some such cells, lying in lamina IV, were similar to spinocervical tract cells and may have projected to both lateral cervical and dorsal column nuclei. Others, at the extreme lateral edge of the mid-dorsal horn, were quite different, with dendrites greatly extended rostrocaudally and primary and higher order dendrites projecting ventrally from the perikaryon.     

Regulation of morphological postsynaptic silent synapses in developing hippocampal neurons

Many excitatory synapses are thought to be postsynaptically 'silent', possessing functional NMDA but lacking functional AMPA glutamate receptors. The acquisition of AMPA receptors at silent synapses may be important in synaptic plasticity and neuronal development. Here we characterize a possible morphological correlate of silent synapses in cultured hippocampal neurons. Initially, most excitatory synapses contained NMDA receptors, but only a few contained detectable AMPA receptors. Synapses progressively acquired AMPA receptors as the cultures matured. AMPA receptor blockade increased the number, size and fluorescent intensity of AMPA receptor clusters and rapidly induced the appearance of AMPA receptors at 'silent' synapses. In contrast, NMDA receptor blockade increased the size, intensity and number of NMDA receptor clusters and decreased the number of AMPA receptor clusters, resulting in an increase in the proportion of 'silent' synapses. These results suggest that the number of silent synapses is regulated during development and by changes in synaptic activity.     

Peripheral vibration causes an adenosine-mediated postsynaptic inhibitory potential in dorsal horn neurons of the cat spinal cord.

We have previously reported a vibration-induced, adenosine-mediated inhibition of nociceptive dorsal horn neurons in the cat spinal cord. The present study was conducted to investigate the mechanisms of this inhibition. In vivo intracellular recording was obtained from dorsal horn neurons in the lower lumbar segments of the anaesthetized cat. Vibration (80-250 Hz for 2-3 s every 15-20 s) was applied to the glabrous skin of the toes of the hind foot using a feedback-controlled mechanical stimulator. In 32 of 43 neurons tested, vibration produced a pronounced hyperpolarization of the membrane potential. This hyperpolarization peaked at -10 mV and decayed throughout the period of the application of vibration. It was associated with a decrease in membrane resistance, had a reversal potential negative to the resting membrane potential and was Cl(-)-independent, suggesting that it was due to an increase in a K+ conductance, properties typical of the response to adenosine. This inhibitory postsynaptic potential was unaffected by intravenous administration of bicuculline, strychnine and naloxone but was blocked by iontophoretic administration of 8-sulphophenyltheophylline, a P1-purinergic receptor antagonist. These results confirm our previous finding that vibration-induced inhibition of nociceptive dorsal horn neurons is mediated via the release of an endogenous purine compound and further suggests that this inhibition involves a postsynaptic inhibitory mechanism.     

Processing of vibrotactile inputs from hairy skin by neurons of the dorsal column nuclei in the cat

The capacity of single neurons of the dorsal column nuclei (DCN) for coding vibrotactile information from the hairy skin has been investigated in anesthetized cats to permit quantitative comparison first with the capacities of DCN neurons responding to glabrous skin vibrotactile inputs and second with those of spinocervical tract neurons responding to vibrotactile inputs from hairy skin. Dynamically sensitive tactile neurons of the DCN the input of which came from hairy skin could be divided into two classes, one associated with hair follicle afferent (HFA) input, the other with Pacinian corpuscle (PC) input. The HFA-related class was most sensitive to low-frequency (<50 Hz) vibration and had a graded response output as a function of vibrotactile intensity changes. PC-related neurons had a broader vibrotactile sensitivity, extending to > or =300 Hz and appeared to derive their input from the margins of hairy skin, near the footpads, or from deeper PC sources such as the interosseous membranes or joints. HFA-related neurons had phaselocked responses to vibration frequencies up to approximately 75 Hz, whereas PC neurons retained this capacity up to frequencies of approximately 300 Hz with tightest phaselocking between 50 and 200 Hz. Quantitative measures of phaselocking revealed that the HFA-related neurons provide the better signal of vibrotactile frequency up to approximately 50 Hz with a switch-over to the PC-related neurons above that value. In conclusion, the functional capacities of these two classes of cuneate neuron appear to account for behavioral vibrotactile frequency discriminative performance in hairy skin, in contrast to the limited capacities of vibrotactile-sensitive neurons within the spinocervical tract system.     

Somatic noxious mechanical stimulation induces Fos expression in the postsynaptic dorsal column neurons in laminae III and IV of the rat spinal dorsal horn

This study was conducted to ascertain the possible expression of Fos-like immunoreactivity (Fos-LI) in the postsynaptic dorsal column (PSDC) neurons in response to noxious mechanical stimulation of the forepaw glabrous area of normal rats. For this purpose, Fos immunohistochemistry along with Fluoro-Gold (FG) retrograde tracing was utilized. After repeated noxious pinching of the forepaw glabrous area, there was a marked increase in number of Fos-LI neurons in the dorsal horn, including Rexed's laminae III and IV, at C5-T1 spinal cord segments ipsilateral to the stimulation. Between segments C5 and T1, about 40% of the Fos-LI neurons in laminae III and IV were distributed at segment C7. In the rats subjected to the noxious pinch coupled with FG injection into the right cuneate nucleus, PSDC neurons double labeled with Fos and FG were localized in the ipsilateral laminae III and IV extending from segment C5 to T1, with about 70% of them distributed at segments C6 and C7. At segment C6 or C7, double-labeled neurons made up about 10% of the PSDC neurons that projected their axons to the cuneate nucleus. Most of the double-labeled neurons appeared fusiform with their primary dendrites projected dorso-ventrally. The present results suggest that the morphologically distinct, subclasses of PSDC neurons in spinal laminae III and IV may contribute to the central transmission of mechanical nociceptive information through the dorsal column into the cuneate nucleus.     

Doubly projecting neurons of the dorsal column nuclei

In a sample of 1700 neurons recorded from the dorsal column nuclei of the cat, 44 (2.6%) were found to send an axon down the dorsal spinal cord. Fully 70% of these caudally projecting neurons also sent an axon to the ventral thalamus. Nearly all had small cutaneous receptive fields distally on the forelimb and displayed response properties similar to other neurons of the dorsal column nuclei. Most were isolated along the lateral and medial margins of the cuneate nucleus rostral to the obex, and many were excited or inhibited by pericruciate cerebral stimulation. A few clearly were excited monosynaptically from the contralateral cerebral cortex at a latency that required the largest pyramidal tract fibers. These neurons probably comprise an important subset that regulates the flow of sensory information in spinal and brainstem somatic sensory pathways.     

Cells of origin of avian postsynaptic dorsal column pathways.

Whereas in the cervical spinal cord of pigeons lamina IV and medial lamina V neurons are at the origin of postsynaptic pathways to the dorsal column nuclei, lumbar lamina IV neurons do not project substantially beyond the cervical enlargement. There is, however a distinct group of medially located lumbar lamina V neurons which projects ipsilaterally to the dorsal column nuclei.     

Pericruciate cortical neurons projecting to brain stem reticular formation,dorsal column nuclei and spinal cord in the cat

Cells of origin of the pericruciate cortical fibers to the bulbar medial reticular formation, the dorsal column nuclei and the pinal cord in the cat were identified by means of the retrograde axonal transport of horseradish peroxidase. After injection of the enzyme in the dorsal column nuclei or the spinal cord many layer V pyramidal neurons were labeled retrogradely in areas 2, 3 and 4 (see ref. 6), but area 4 giant Betz cells were only labeled after spinal cord injections. Medial bulbar reticular formation injections resulted in the labeling of pyramidal neurons mainly in area 6.     

The fine structure of synapses in relation to the large spherical neurons in the anterior ventral cochlear of the cat

Summary The fine structure of synapses in relation to the large spherical neurons, the principal neurons in the anterior ventral cochlear nucleus (AVCN) of the cat, was studied. Axo-somatic synapses were classified into three types based on their size and shape and the distribution of synaptic vesicles. Axo-dendritic synapses were also classified into three types with respect to synaptic contact, cytoplasmic densities associated with the pre- and postsynaptic membranes and the shape of the synaptic vesicles. Mossy fibre-like endings and preterminal axons containing dense-core vesicles were observed in this study. Serial or triadic synapses were also found in cat AVCN. We occasionally found a nematosome in the cytoplasm of the principal neuron.     

Recovery of function is not associated with proliferation of retinogeniculate synapses after chronic deafferentation in the dorsal lateral geniculate nucleus of the adult cat

A restricted recovery of visual excitation occurs in the partially deafferented dorsal lateral geniculate nucleus (dLGN) of the cat after retinal lesions. In the absence of axonal growth an increase of retinogeniculate synapses at the peripheral dendrites of deafferented cells could be a possible underlying mechanism. We labeled optic tract terminals with horseradish peroxidase and [3H]proline. The size and density of labeled boutons in the vicinity of regions deafferented by chronic retinal lesions were not different from those in normal parts of the dLGN. There was no indication for synaptic proliferation as a response to partial visual deafferentiation in the adult cat dLGN.     

Physiological studies on neurons in the dorsal cochlear nucleus of cat

Results reported here support the conclusion that an individual neuron in the dorsal cochlear nucleus (DCN) can exhibit pauser, buildup, and chopper patterns in response to tone pips. Fusiform cells have been previously identified as the principal cell exhibiting these patterns. Fusiform cells can also exhibit an onset response followed by suppression of spontaneous activity at their characteristic frequency (CF). Off CF only suppression is seen. These neurons are characterized by a restricted excitatory region near threshold. All these cells can exhibit nonmonotonic rate curves, narrow excitatory regions, and inhibitory sidebands. Nonmonotonicity occurred in 34% of pausers, 52% of buildup, 89% of onsets with a graded response, and 50% overall in the DCN cells. Chopper units occur as often as the other types combined in the DCN. Only 14% show nonmonotonic rate curves. Those with high-spontaneous activity also show inhibitory sidebands. Cells with a predominant buildup pattern occur most frequently in the fusiform cell layer, whereas pausers occur throughout the DCN below the molecular layer. Intracellular potentials often reflect the average response pattern. Sharply delimited response areas indicate that these cells may be useful for performing a spectral analysis. These cells show almost no phase locking suggesting that temporal encoding is an unlikely function. It is suggested that the effects of anesthetic on the function of the DCN is not as marked as previously indicated.     

Organization of the serotonergic innervation of spinal neurons in rats—I. Neuropeptide coexistence in varicosities innervating some spinothalamic tract neurons but not in those innervating postsynaptic dorsal column neurons

Previous studies have suggested that peptides such as substance P and thyrotropin-releasing hormone coexist with serotonin in the same varicosities in the ventral horn and intermediate gray of the spinal cord in rat. However, coexistence of these peptides with serotonin is rare in fibers in the superficial dorsal horn. Since it has been proposed that serotonergic fibers in the superficial dorsal horn act to modulate nociception, it was hypothesized that the serotonergic neurons that contain neither substance P nor thyrotropin-releasing hormone might constitute a specifically antinociceptive subset of serotonergic neurons. This being the case, it would be expected that different types of serotonergic neurons innervate nociceptive and non-nociceptive spinal neurons. In order to test this hypothesis, a group of cells that include nociceptive neurons (spinothalamic tract neurons) and a group of predominantly non-nociceptive neurons (postsynaptic dorsal column neurons) in the spinal cord of rat were retrogradely labeled. Sections of the spinal cord containing retrogradely labeled spinothalamic tract or postsynaptic dorsal column neurons were stained for serotonin and either substance P or thyrotropin-releasing hormone using two-color immunohistochemistry. A retrogradely labeled cell was classified as “apposed” if there was no discernible distance between an immunohistochemically labeled varicosity and the cell. Eighty per cent of spinothalamic tract and 83% of postsynaptic dorsal column profiles were apposed by serotonin-immunoreactive varicosities in the spinal cord. Thirty-one per cent of the spinothalamic tract profiles that were apposed by serotonergic varicosities were apposed by serotonergic varicosities that were also stained for thyrotropin-releasing hormone. The distribution of the latter spinothalamic neurons was similar to that reported for spinothalamic tract neurons responsive to joint movement. In addition, at least 63% of the spinothalamic tract profiles which were apposed by serotonergic varicosities were apposed by “serotonin-only” varicosities, including most spinothalamic tract neurons in the marginal zone, suggesting that at least some “serotonin-only” neurons are antinociceptive. However, contrary to the hypothesis, at least 94% of the postsynaptic dorsal column profiles apposed by serotonergic varicosities were apposed by “serotonin-only” varicosities.

These findings suggest that there may be a relationship between the sensory modality to which a spinal neuron responds and the type of serotonergic innervation it receives. However, it appears that “serotonin-only” neurons may not constitute a specifically antinociceptive category of serotonergic neurons.