Cervical Nucleus Research Articles

Glutamate and AMPA receptor immunoreactivity in Ia synapses with motoneurons and neurons of the central cervical nucleus.

Axonal tracing and high resolution immunocytochemistry were used to identify transmitter content and postsynaptic receptors in synapses between Ia primary afferents and motoneurons and in neurons of the central cervical nucleus (CCN), respectively, in the rat. The terminals, as well as the target neurons, were identified by postembedding immunogold detection of transganglionically or retrogradely, respectively, transported cholera toxin B subunit (CTB), and in adjacent sections postembedding immunogold was employed to demonstrate glutamate and AMPA receptors in the same synapses. A total of 390 CTB-labelled Ia boutons in apposition to CTB-labelled motoneurons, CCN neurons or unlabelled dendrites in the surrounding neuropil were traced in section series from two animals. A third animal was used as a control. In the motor nucleus, a majority of the synapses were with medium-sized dendrites, whereas in the CCN the distribution was skewed towards fine-calibre dendrites. In both nuclei, somatic and juxtasomatic synapses were quite infrequent (<10%). All of the CTB-labelled Ia boutons recovered in the sections incubated for glutamate (n=323) were enriched with glutamate immunoreactivity. One hundred and fifty of these disclosed synaptic contact in at least two ultrathin sections. In this sample, 50% (33-59%) appeared immunoreactive to receptor sub-units GluR1-4 in at least two ultrathin sections, whereas 35% were labelled in one section only. Distribution of gold particles relative to presynaptic and postsynaptic membrane profiles (n=23) revealed a close correlation between AMPA immunoreactivity and the postsynaptic membrane of the synapse. Finally, immunogold particles signalling GluR1 were observed much less frequently than particles signalling GluR2/3 or GluR4. Our results provide additional strong evidence that chemical transmission at Ia synapses is mediated by glutamate and identify GluR2/3 and GluR4 as important postsynaptic receptors.

GABA-, glycine-, and glutamate-immunoreactive bouton profiles in apposition to neurons of the central cervical nucleus in the rat.

The neurons of the central cervical nucleus (CCN) convey information about the position and movements of the head, and receive excitatory input from dorsal neck muscles and the labyrinth. Both of these afferent sources form glutamatergic synaptic contacts with CCN neurons. However, these sensory afferent sources can also inhibit CCN neurons. To further elucidate the synaptic organization, we made an electron microscopic investigation, identifying and evaluating the relative frequency of bouton profiles containing the inhibitory transmitters GABA and glycine in apposition to identified CCN neurons. In addition, labeling for glutamate was performed. The identification of the CCN neurons was made possible by injections of retrograde tracer substances into the cerebellum. These substances were made visible by preembedding immunocytochemistry or postembedding immunogold staining. Such staining was also used to detect the three amino acids that were found in boutons apposed to the identified neurons (cf. Ornung et al., J. Comp. Neurol. 1996;365:413-426; Lindå et al., J. Comp. Neurol. 2000;425:10-23). Due to the relatively poor transport of the tracer substances into dendrites of the CCN neurons, the analysis was restricted to the cell body and included bouton profiles in direct apposition to the soma membrane. Data from 10 CCN neurons revealed that about 50% of the apposing bouton profiles were immunoreactive for GABA, and about 34% for glycine. In four neurons, the degree of colocalization of GABA and glycine was determined to be close to 30%. Thus, the vast majority of glycine-labeled profiles also contained GABA, while a considerable fraction of the profiles were immunoreactive for only GABA. The values for glycine immunoreactive bouton profiles presented here may represent somewhat low estimates, depending on the method used. Data from four neurons showed that about 18% of the profiles were labeled for glutamate. The large fraction of purely GABA immunoreactive profiles, or at least a substantial group of them, is suggestive of their derivation from axons descending from the brainstem.

Organization of the ferret lateral cervical nucleus and cervicothalamic tract

To elucidate the organization of the ferret spinocervicothalamic pathway (SCTP), we examined the lateral cervical nucleus (LCN) and the termination of the cervicothalamic tract (CTT) in this species. In thionin-stained sections, the ferret LCN appeared as an easily delineated column of cells in the dorsolateral funiculus from about mid-C3 to the rostral end of C1, with most cells located in the C1 and C2 segments. In transverse sections, the LCN was elongated along a dorsolateral to ventromedial axis and in the rostral half of C2 and caudal half of C1 continuous with the neck of the dorsal horn. The number of ferret LCN cells was estimated to 2,500-3,700, with an average of 3,340. Substance P-like immunoreactive fibers located preferentially in the ventromedial part of the LCN, whereas serotonin-like immunoreactive fibers were found throughout the nucleus. Anterograde transport of wheat germ agglutinin-horseradish peroxidase conjugate and biotinylated dextran amine demonstrated that the ferret CTT terminates extensively in the peripheral parts of the ventral posterior lateral nucleus. Sparser termination was evident in the ventral posterior inferior nucleus, in the medial nucleus of the posterior complex, and in the medial part of the magnocellular medial geniculate nucleus. Thus, although the LCN is significantly smaller in ferrets than in cats and raccoons, the organization of the LCN and of the cervicothalamic tract is closely similar in the three species. These findings indicate a conserved general organization of the SCTP among carnivores.

Uncrossed and crossed projections from the upper cervical spinal cord to the cerebellar nuclei in the rat, studied by anterograde axonal tracing.

In the upper cervical spinal segments, neurons in the medial part of lamina VI give rise to uncrossed spinocerebellar axons, whereas the central cervical nucleus (CCN) and neurons in laminae VII and VIII give rise to crossed spinocerebellar axons. Using anterograde labeling with biotinylated dextran in the rat, we examined the projections of these neuronal groups to the cerebellar nuclei. Uncrossed and crossed projections were distinguished by cerebellar lesions placed on the side contralateral or ipsilateral to the tracer injections confined to the second and third cervical spinal segments (C2 and C3, respectively). Labeled terminals of uncrossed projections were seen in the middle, dorsal, and ventrolateral parts of the middle subdivision and in the ventral part of the caudomedial subdivision of the medial nucleus. In the anterior interpositus nucleus, terminals were seen in the middle of the mediolateral extent, whereas, in the posterior interpositus nucleus, they were seen in lateral and caudal parts. The terminals of crossed projections from the CCN were distributed ventrally in medial to ventrolateral parts of the middle subdivision of the medial nucleus. Some terminals were seen in the caudomedial subdivision of the medial nucleus. In the anterior interpositus nucleus, labeled terminals were seen mainly in rostromedial parts, whereas, in the posterior interpositus nucleus, they were seen in caudal and dorsal parts of the medial half. The present study suggests that the medial lamina VI group and the CCN in the upper cervical segments project to the different areas of the cerebellar nuclei and are concerned with different functions.

The organization of the spinocerebellar tract neurons in the chicken

The organization of spinocerebellar tract (SCT) neurons in the chicken was studied quantitatively using retrograde wheat germ agglutinin-horseradish peroxidase labelling. The chicken spinal cord was divided into five regions based on the distribution of SCT neurons: the cervical region (the spinal segments [SS], 1–12, which contained 32% of the total number of SCT neurons), the cervical enlargement (SS13–15, 13.4%), the thoracic region (SS16–20, 13%), the thoraco-lumbosacral region (SS21–26, 34.6%), and the posterior lumbosacral region (SS27–30, 7%). Clarke’s column was found in two regions, a cervical one (SS12–16) and a lumbar one (SS21–28). The spinal border cells were less numerous and also present in two parts, a cervical one (SS10–15) and a caudal one (SS20–24). SCT neurons of the ventral part of the ventral horn were found in the cervical enlargement and SS16 (lamina VIII), and in the thoraco- and posterior lumbosacral regions (laminae VIII and IX). The ventral marginal nucleus consisted exclusively of SCT neurons but the major and minor marginal nuclei did not contain SCT neurons. In the cervical region most of SCT neurons were observed in the ventral horn, while the central cervical nucleus, which consists of SCT neurons in mammals, was not identified in the chicken.

Projection Linkage from Spinal Neurons to Both Lateral Cervical Nucleus and Solitary Tract Nucleus in the Cat

Intracellular recordings from the lumbosacral dorsal horn were made to identify the axonal projection and the afferent innervation of the lateral cervical nucleus (LCN) and solitary tract nucleus (STN) on the spinal neurons of chloralose-anesthetized cats. A total of 49 neurons from laminae III–V in the spinal dorsal horn responded to stimulation of both the LCN and STN. Of these, 28 and 21 neurons responded antidromically and orthodromically to stimulation of the LCN and STN, respectively. Seven of the 28 antidromically activated neurons were followed by one or more responses synaptically driven from the LCN and/or STN. The diameter of these ascending or descending fibers was in the range of A delta fibers. The results indicate that (1) some spinal neurons, namely spinocervical tract-spinosolitary tract (SCT-SST) neurons, issue branched axons of A delta-fibers and dually project to both LCN and STN; (2) some SCT-SST neurons receive innervation from both the LCN and STN; (3) some spinal neurons and interneurons are dually innervated by descending fibers originating from both the LCN and STN, and (4) the convergence and integration between somatic and visceral sensory inputs might occur in the SCT-SST neurons.

Background activity of rabbit hippocampal neurons in conditions of functional exclusion of structures which regulate the theta rhythm.

The functional importance of theta modulation in the activity of hippocampal neurons was further analyzed using a method consisting of controlled sequential short-term (25-30 min) inclusion or exclusion of the theta rhythm by local administration of lidocaine into the median cervical nucleus and medial septal region respectively. Studies were carried out using conscious rabbits with extracellular recording of hippocampal neuron activity in field CA1. Administration of lidocaine into the medial septal nucleus and diagonal tract nucleus (MS-DT) led to complete inhibition of theta modulation in neuronal and total hippocampus activity. The mean frequency of background discharges underwent no change in most neurons, but decreased significantly in a limited group of cells with high-frequency activity (presumptive inhibitory neurons). Administration of lidocaine into the median cervical nucleus (MCN), the source of serotoninergic pathways to the MS-DT and hippocampus, was accompanied by increases in the stability and frequency of theta modulation of neuronal activity, induction of theta modulation in an additional group of neurons, and expression of a continuous theta rhythm in the electrical activity (EA) of the hippocampus. The mean frequency and regularity of discharges increased in most cells. These data support the existence of tonic inhibitory effects on the part of the MCN on the septa-hippocampal system generating the theta rhythm; in this regard, the MCN can be regarded as an antagonist of the activating reticular formation.

Origin, course, and laterality of spinocerebellar axons in the North American opossum, Didelphis virginiana.

Spinocerebellar axons have been studied extensively in placental mammals, but there have been no full reports on their origin, laterality, or spinal course in any marsupial. We have used the North American opossum (Didelphis virginiana) to obtain such information and to ask whether any spinocerebellar neurons innervate both the anterior and posterior lobes of the cerebellum through axonal collaterals. To identify spinal neurons that project to the cerebellum, we employed the retrograde transport of Fluoro-Gold (FG) from the anterior lobe, the main target of spinocerebellar axons. In some cases, cerebellar injections of FG were combined with hemisections of the rostral cervical or midthoracic spinal cord, so that laterality of spinocerebellar connections could be established. To determine whether single neurons project to both the anterior lobe and the posterior lobe, injections of Fast Blue (FB) into the anterior lobe were combined with injections of Diamidino yellow (DY) or rhodamine B dextran (RBD) into the posterior lobe, or vice versa. Following injections of FG into the anterior lobe, neurons were labeled throughout the length of the spinal cord, which differed in laminar distribution and laterality of their projections. Among other areas, neurons were labeled in the central cervical nucleus, the nucleus centrobasalis, Clarke's nucleus, the dorsal horn dorsal spinocerebellar tract area, the spinal border region, and Stilling's nucleus. When anterior lobe injections of FB were combined with injections of RBD or DY into the posterior lobe, or vice versa, some double-labeled neurons were present in all major spinocerebellar groups. Cerebellar injections of FG also retrogradely labeled spinocerebellar axons, allowing us to document their locations in the gray matter as well as within the periphery of the lateral and ventral funiculi at all spinal levels. A few spinocerebellar axons also were found in the dorsal funiculus (a dorsal column-spinocerebellar tract), which appeared to originate from neurons in the dorsal part of Clarke's nucleus from the ninth thoracic segment to the first lumbar segment. Our results indicate that spinocerebellar axons in the marsupial opossum are generally comparable in origin, course, and laterality to the same axons in the placental mammals studied to date.

Ultrastructural detection of neuronally transported choleragenoid by postembedding immunocytochemistry in freeze-substituted Lowicryl HM20™ embedded tissue

Choleragenoid (cholera toxin B-fragment; CTB) is an anterograde, retrograde and transganglionic neuronal tracer. We describe a method for detecting CTB-labeled neuronal cell bodies, neurites and boutons at the ultrastructural level, using postembedding immunogold techniques on freeze-substituted Lowicryl HM20™ embedded nervous tissue. Primary afferents and motoneurons were labeled by injection of CTB in the dorsal ramus of the C2 spinal nerve of the rat. Following fixation with paraformaldehyde (4%) and glutaraldehyde (0.25%), tissue sections from the spinal cord C2 segment were freeze-substituted and embedded in Lowicryl HM20™ and subsequently processed with postembedding immunocytochemistry for CTB and glutamate. Immunogold particles indicating CTB immunoreactivity were found over primary afferents and motoneurons. In primary afferents in the central cervical nucleus (CCN) and motor nuclei, immunogold labeling was seen in boutons over vesicle-containing axoplasm and to a lesser extent over axoplasm devoid of vesicles, but not over mitochondria or axolemma. In motoneurons, immunogold particles were seen over the Golgi apparatus in the soma and over lysosomes in both soma and dendrites. Quantification of glutamate-like immunoreactivity in 20 CTB-labeled and 20 CTB-negative boutons in the neuropil was found similar, indicating that CTB does not interfere with the immunocytochemical detection of neuronal epitopes such as the transmitter substance glutamate.

Neuropeptide FF in the lateral spinal and lateral cervical nuclei: evidence of contacts on spinothalamic neurons.

Neuropeptide FF (NPFF, F8Famide) is best known for its modulating effect on opioid analgesia and morphine tolerance. However, the exact mode of action of NPFF in sensory transmission is not known. We compared the distribution of NPFF-immunoreactive (ir) fibers and terminal-like thickenings with the retrograde, tracer-filled spinothalamic (ST) neurons in the lateral spinal nucleus (LSN) and lateral cervical nucleus (LCN) of rat, areas where NPFF-containing nerve terminals are abundant. We injected fluorescent latex microspheres into the ventroposterolateral thalamic nucleus and more medial thalamic nuclei, which are innervated by ST neurons. We found NPFF-ir terminal-like thickenings and fibers apposing the tracer-filled neurons in the LSN and LCN. ST neurons filled with the retrograde tracer making contacts with NPFF-ir terminal-like thickenings, were found to terminate not only in the ventroposterolateral thalamic nucleus but also in more medial thalamic nuclei. The highest number of tracer-filled ST neurons having NPFF-ir terminal-like thickenings and fibers in apposition were found at the cervical level. Our results suggest that NPFF-containing systems in the spinal cord of rat are not limited to the substantia gelatinosa, and the sensory functions of NPFF may be mediated at least partly through the modulation of the ST system. NPFF-ir contacts in the LSN and LCN might play an important role in the somatic sensory transmission system. This study shows evidence for the first time that the spinal NPFF-containing system may be involved in mechanisms that control sensory input to the supraspinal levels.

Indications for coupling between feline spinocervical tract neurones and midlumbar interneurones.

The possibility of collateral segmental actions of spinocervical tract (SCT) neurones upon interneurones with input from cutaneous and group II muscle afferents was investigated in deeply anaesthetized cats. To this end, intracellular and/or extracellular recordings were made from 35 dorsal horn and 15 intermediate zone interneurones in midlumbar segments of the spinal cord and effects of stimulation of the ipsilateral dorso-lateral funiculus (DLF) at C3 and C1 levels, i.e. below and above the lateral cervical nucleus where axons of SCT cells terminate, were compared. The stimuli applied at the C3 segment were within the range of stimuli (50-100 microA) required for antidromic activation of SCT neurones in the same experiment. Those applied at the C segment (200-500 microA) were at least 3 times stronger than C3 stimuli. Under the same experimental conditions, long ascending and descending tract neurones (dorsal spino-cerebellar and rubro-spinal tract neurones) with axons in the DLF were activated at similar thresholds from the C and C3 segments. Intracellular recordings were made from 29 interneurones of which 19 (65%) were dorsal horn and 10 (35%) were intermediate zone interneurones. Excitatory postsynaptic potentials (EPSPs) evoked by single stimuli applied at the C3 segment, but not the C segment, were found in 14 (48%) of those interneurones; their latencies (3.0-5.7 ms) and frequency following with only minimal temporal facilitation were as required for potentials being evoked monosynaptically by the fastest conducting SCT neurones. Extracellular recordings were made from 30 interneurones (24 dorsal horn and 6 intermediate zone interneurones), and in these neurones spike potentials induced from the C3, but not from the C segment, were evoked only by short trains of stimuli. However, their latencies from the first effective stimulus (4.3-5.4 ms) were compatible with mono- or oligosynaptically mediated collateral actions of SCT neurones. They were found in 10 (33%) of the 30 investigated interneurones. Similar effects of C3 stimuli were found in similar proportions of dorsal horn interneurones and intermediate zone interneurones. Indications were also found for synaptic actions evoked by C3 stimuli that could not be attributed to direct collateral actions of SCT neurones. In some intracellularly recorded dorsal horn interneurones, short-latency EPSPs were evoked from the C3 segment by the 2nd or 3rd stimulus in the train, but not by single stimuli. In other dorsal horn and intermediate zone interneurones, inhibitory postsynaptic potentials (IPSPs) were evoked from the C3 segment at minimal latencies (2.7-3.2 ms), which might be too short to allow their mediation via SCT neurones. We conclude that SCT neurones might be used to forward information from muscle group II and cutaneous afferents not only to neurones in the lateral cervical nucleus and via them to thalamus and cerebral cortex but also to interneurones in spinal reflex pathways. Thereby reflex actions evoked from group II and cutaneous afferents might be co-ordinated with responses mediated by supraspinal neurones. We conclude also that dorsal horn and intermediate zone mid-lumbar interneurones might contribute to the previously reported di-and poly-synaptic excitation or inhibition of postsynaptic dorsal column (PSDC), spinothalamic tract (STT) and spinomesencephalic tract (SMT) neurones by collateral actions of SCT cells. Thereby these interneurones might contribute to the co-ordination of responses mediated by various populations of supraspinal neurones.

Electrophysiological evidence that spinomesencephalic neurons in the cat may be excited via spinocervical tract collaterals

Extracellular microelectrode recordings were made from spinomesencephalic tract (SMT) neurons in the lumbosacral spinal cord of cats anaesthetized with chloralose and paralysed with gallamine triethiodide. The SMT cells were antidromically fired from the posterolateral parts of the superior colliculus and the intercollicular region, were located in laminae IV to VIII, and had response properties and axonal conduction velocities similar to those described previously. The effects of stimulating the dorsolateral funiculus of the cervical cord at C3 and rostral C1, below and above the termination of spinocervical tract (SCT) axons in the lateral cervical nucleus, were examined on 33 SMT cells. The strength of stimulation was adjusted so that at C3 it was above threshold for antidromic activation of SCT cells and at C1 was below threshold for activation of the same cells. Seven (21%) SMT neurons were excited from C3 but not from C1. The remaining 26 (79%) were excited from both C3 and rostral C1 and 23 (70% of these) were excited significantly more from C3. That is, 91% of the total sample were either excited only from C3 or more strongly from C3 than from rostral C1. We discuss the possible neuronal systems involved and conclude that the greater excitatory effects from C3 are most likely due to antidromic activation of the SCT. The shortest latency effects from C3 indicate a monosynaptic linkage between SCT cells with the fastest axons and the SMT. The longer latency actions may be due to monosynaptic connexions from SCT cells with slower conducting axons, to di- or polysynaptic actions from SCT cells with fast axons, or a combination of both. SMT cells are another population of spinal neurons, in addition to postsynaptic dorsal column, spinothalamic and dorsal horn spinocerebellar neurons, which receive excitation via SCT collaterals.

Excitatory connections between neurons of the central cervical nucleus and vestibular neurons in the cat.

The central cervical nucleus (CCN) of the cat receives input from upper cervical muscle afferents, particularly primary spindle afferents. Its axons cross in the spinal cord, and while in the contralateral restiform body give off collaterals to the vestibular nuclei. In order to study the connections between CCN axons and vestibular neurons, we stimulated the area of the CCN in decerebrate cats while recording intra- or extracellularly from neurons in the contralateral vestibular nuclei. CCN stimulation evoked excitatory postsynaptic potentials (EPSPs) or extracellularly recorded firing in the lateral, medial and descending vestibular nuclei. The latency of EPSPs (mean 1.6 ms) was on average 0.4 ms longer than the latency of antidromic spikes evoked in the CCN by stimulation of the contralateral vestibular nuclei (mean 1.2 ms), demonstrating that the excitation was typically monosynaptic. The results provide further evidence that the CCN is an important excitatory relay between upper cervical muscle afferents and neurons in the contralateral vestibular nuclei.

Spinal ascending pathways in amphibians: Cells of origin and main targets

As part of a research program on the evolution of somatosensory systems in vertebrates, the various components of ascending spinal projections were studied with in vivo and in vitro tract-tracing techniques in representative species of amphibians (the large green frog, Rana perezi, the clawed toad, Xenopus laevis and the ribbed newt, Pleurodeles waltl). Three main ascending sensory channels, each with largely separate targets, were demonstrated: 1. Ascending projections via the dorsal funiculus include primary and nonprimary projections that ascend to terminate mainly in the dorsal column nucleus at obex levels. A small component ascends farther rostralwards to terminate in the reticular formation, the octavolateral area, the trigeminal nuclear complex, and in the granular layer of the cerebellum. 2. Projections ascending via the dorsolateral funiculus reach other spinal and supraspinal targets than the dorsal funicular fibers, mainly ipsilaterally. At upper cervical cord and obex levels, many fibers innervate a region considered the amphibian homologue of the lateral cervical nucleus of mammals. In the medulla, these fibers ascend ventral to the descending trigeminal tract to terminate in the dorsal column and the solitary tract nuclei, and more rostrally, in the reticular formation, the descending trigeminal nucleus and the medial aspect of the ventral octaval nucleus. Major projections reach the area between the facial motor nucleus and the ventral octaval nucleus, and a mediolateral subcerebellar band. These projections arise in neurons located mainly in the ipsilateral deep dorsal and lateral fields throughout the spinal cord. 3. Ascending spinal projections via the ventral quadrant of the spinal cord (the ventral and ventrolateral funiculi) ascend throughout the brainstem up to the diencephalon. Along its course, this component innervates various parts of the reticular formation, the octavolateral area, the granular layer of the cerebellum, the region ventromedial and ventrolateral to the isthmic nucleus, and the subcerebellar region. In the mesencephalon, the torus semicircularis, the midbrain tegmentum and, sparsely, the tectum mesencephali are innervated. Beyond the midbrain, various dorsal and particularly ventral thalamic nuclei and the posterior tubercle are innervated by this ascending sensory channel. The cells of origin of some of these projections were observed in the dorsal, and to a lesser extent, in the lateral and ventral spinal fields of the spinal cord. Evidence for the presence of these three main ascending sensory channels throughout vertebrates will be discussed. The presence of such channels appears to be a shared character in the brain of both amniotes and anamniotes.

Nerve growth factor concentrations in human cerebral blood vessels.

The distribution of glutamate-like immunoreactivity (Glu-LI) in the thalamic ventral posterolateral nucleus (VPL) of cats was studied with the EM immunogold technique in order to identify nerve terminal populations that...

1502 Connections between neurons of the central cervical nucleus and vestibular neurons in the cat

1502 Connections between neurons of the central cervical nucleus and vestibular neurons in the cat

Connections between neurons of the central cervical nucleus and vestibular neurons in the cat

The primary regions affected by descending projections of the pedunculopontine nucleus (PPN) are the subcoeruleus (SubC) nucleus, the pontine and medullary reticular formation, and, to a lesser extent, the spinal cord. The SubC modulates rapid eye movement (REM) sleep signs such as ponto–geniculo–occipital (PGO) waves, atonia, and outputs to the hippocampus. The pontine and medullary reticular formation controls reticulospinal pathways involved in locomotion and postural control. Descending projections to the spinal cord are sparse and their sites of termination have not been described. However, descending projections of the reticular activating system (RAS) are critical for the control of postural and locomotor events related to waking as well as to REM sleep.

Cyto- and myeloarchitectonic organisation of the spinal cord of an echidna (Tachyglossus aculeatus).

We have studied the cyto- and myeloarchitectural organisation of the spinal cord of an echidna (Tachyglossus aculeatus) with the aid of Nisst staining, darkfield examination and p-phenylenediamine staining. We have also examined the distribution of unmyelinated afferents by labelling with a peroxidase-conjugated lectin derived from Griffonia simplicifolia (B1 isolectin). The cytoarchitectural features characterising the laminar organisation of the spinal cord in eutherian mammals were broadly applicable to the spinal cord of this monotreme. In addition, we identified a distinct group of large neurons in the ventral part of lamina X, extending into the ventral funiculus, that we have called the median nuclear group. We were unable to identify a central cervical nucleus in this echidna on the basis of cytoarchitectural criteria, although all other spinal cord nuclei found in eutherians could be found in this monotreme. Lectin labelling with the Griffonia simplicifolia isolectin B4 revealed a subpopulation of dorsal root ganglion cells similar to those labelled in Eutheria. In this echidna, labelling of unmyelinated fibres was found in Lissauer's zone and laminae I and II, as seen in rats (Rattus norvegicus); there were also deeper patches extending into laminae III to V and what appeared to be commissural axons approaching the dorsal grey commissure, which have not been seen in Eutheria. Fibre calibre in the dorsal and ventral roots of this echidna was similar to that reported in Eutheria, suggesting similar proportion of afferent fibre classes and alpha and gamma motoneurons. In the echidna, mean diameter of myelinated dorsal root axons was 4.65 microns at T1 and 5.22 microns at L3, with a clear bimodal distribution in the L3 dorsal root showing distinct groups at 1 to 5 microns and 6 to 12 microns. These made up approximately 45 and 55% of the total myelinated axon population, respectively. Myelinated fibres in the ventral root at L3 showed two major peaks in distribution. These were at 1 to 4 microns (approximately 32% of the total myelinated fibre population) and at 7 to 14 microns (approximately 58% of the total myelinated fibre population). The cross-sectional area of the dorsal columns of this monotreme was comparable to that of a eutherian mammal of similar body weight, and myelinated axon calibre was similar to that seen in the domestic cat. Our findings indicated that spinal cord cytoarchitectural organisation is highly conserved across class Mammalia, although the lectin labelling findings suggested that termination of unmyelinated afferents in echidnas may differ from that found in Eutheria. The dorsal column system appears to be as anatomically well developed in this monotreme as in those eutherian mammals considered to have a pronounced discriminative tactile sense.

Distribution of dopamine immunoreactivity in the rat, cat and monkey spinal cord.

In the present study, the distribution of dopamine (DA) was identified light microscopically in all segments of the rat, cat, and monkey spinal cord by using immunocytochemistry with antibodies directed against dopamine. Only fibers and (presumed) terminals were found to be immunoreactive for DA. Strongest DA labeling was present in the sympathetic intermediolateral cell column (IML). Strong DA labeling, consisting of many varicose fibers, was found in all laminae of the dorsal horn, including the central canal area (region X), but with the exception of the substantia gelatinosa, which was only sparsely labeled, especially in rat and monkey. In the motoneuronal cell groups DA labeling was also strong and showed a fine granular appearance. The sexually dimorphic cremaster nucleus and Onuf's nucleus (or its homologue) showed a much stronger labeling than the surrounding somatic motoneurons. In the parasympathetic area at sacral levels, labeling was moderate. The remaining areas, like the intermediate zone (laminae VI-VIII), were only sparsely innervated. The dorsal nucleus (column of Clarke) showed the fewest DA fibers, as did the central cervical nucleus, suggesting that cerebellar projecting cells were avoided by the DA projection. In all species, the descending fibers were located mostly in the dorsolateral funiculus, but laminae I and III also contained many rostrocaudally oriented fibers. It is concluded that DA is widely distributed within the spinal cord, with few differences between species, emphasizing that DA plays an important role as one of the monoamines that influences sensory input as well as autonomic and motor output at the spinal level.

Regulation of the septal pacemaker theta rhythm by the cervical nuclei of the midbrain.

Neuronal activity in the medial septal region (the medial nucleus and the diagonal band nucleus, MN-DBN) was recorded along with hippocampal EEG traces in conscious rabbits with stimulatory electrodes implanted in the median cervical nucleus (MCN) and the reticular formation (RF) of the midbrain and pons. In all animals with electrodes in the MCN, the background theta activity frequency was low (4.6 +/- 0.15 Hz) as compared with intact rabbits or those with electrodes implanted only in the RF (5.2 +/- 0.19 Hz, p < 0.5). Stimulation of the MCN with weak low-frequency impulses reduced theta volleys from MN-DBN cells, reducing their frequency and regularity and inducing the appearance or strengthening of low-frequency delta modulation. The number of spikes in a volley decreased, and the duration of inter-volley intervals increased. Stimulation of the MCN led to a gradual decrease in the frequency and amplitude of theta waves, induced irregular delta waves and spindles of 12 Hz in the hippocampal EEG. Stimulation of the RF produced the opposite changes in volley activity in the MN-DBN and hippocampal EEG, with increases in theta and decreases in delta components. These results support a role for the midbrain cervical nuclei as structures limiting the generation of theta activity by the reticular-septal system, but do not support the existence of an MN-DBN-independent high-frequency serotoninergic theta rhythm. It is proposed that the effect of the MCN may be important for suppression and switching of attention.