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Notes: Abstract This report deals with an ultrastructural investigation of the synapses of the proximal axons of normal-appearing anterior horn neurons of 7 patients with amyotrophic lateral sclerosis (ALS) and 4 patients with motor neuron disease who had no upper motor neuron and corticospinal tract involvement (lower motor neuron disease, LMND). Specimens from 12 age-matched individuals who died of non-neurological diseases served as controls. Proximal axons directly emanating from the normal-appearing neurons were examined: 42 axons were from ALS patients, 43 from LMND patients and 87 from controls. Our results show that the number of synapses on axon hillocks, as well as the lengths of the synaptic contact and of the active zone were reduced in both groups of patients (P 〈 0.0001), but no significant differences were seen between patients and controls with respect to the synaptic parameters of initial axon segments. There was no overall difference between ALS and LMND patients. These findings suggest that the electrophysiological functions pertaining to integration of electrical inputs into the axon and information transduction on the axon may be greatly impaired in the early stages of motor neuron diseases, and that the observed synaptic alterations may be pathological events, likely to be due to anterior horn neuron degeneration.

Notes: Abstract This ultrastructural study deals with the synapses of primary dendrites of the anterior horn neurons in the lower lumbar spinal cords of seven patients with amyotrophic lateral sclerosis (ALS) who had mild neuronal depletion. Specimens from seven age-matched, neurologically normal individuals served as controls. In each instance, the autopsy was performed within 6 h after death. Our results indicate a statistically significant increase in degenerative changes in the dendrite presynapses of normal-appearing neurons of the ALS patients. The alterations included aggregation of electron-dense synaptic vesicles and dark mitochondria with dense cristae, and bundles of neurofilaments. However, we found no significant difference between controls and patients with respect to cross-sectional area and length of the dendrites, number of synapses per dendrite, and lengths of individual synapses and their active zones in the normal-appearing neurons, even though the patients' neurons had a smaller cross-sectional area. In chromatolytic neurons, the number of synapses and the length of the active zone of the primary dendrites were significantly diminished. These findings suggest that despite degenerative changes of the presynapses, the synapses in the primary dendrites of the anterior horn neurons are preserved at the early stage of ALS. The preservation of these synapses may be due to their relative resistance to degenerative processes, or may represent a compensatory mechanism of the synapses for diminished synaptic function in distal dendrites.

Notes: Abstract This report concerns an ultrastructural investigation of the synapses of anterior horn neurons in the lumbar spinal cord of four patients with lower motor neuron disease (LMND) who had no upper motor neuron and corticospinal tract involvement. Anterior horn neurons of five normal individuals served as controls. The cell body area and the number of synapses of the normal-appearing neurons of the LMND patients were significantly reduced (P 〈 0.0001). These findings suggest that synaptic changes of anterior horn neurons could be ascribed to the degeneration of lower motor neurons rather than to the influence of upper motor neuron system degeneration. On the other hand, the lengths of individual synapses (P 〈 0.0001) and of their active zones (P 〈 0.05) were significantly increased in the patients. These increases would indicate that synapses on anterior horn neurons of individuals with LMND appear to have the capacity to react to progressive degeneration and loss of other synapses by means of a compensatory response or plasticity that enhances their efficiency.

Notes: Abstract This report deals with an ultrastructural investigation of the synapses of anterior horn neurons in the lumbar spinal cords of five patients with amyotrophic lateral sclerosis (ALS) who had mild neuronal depletion. Specimens from five age-matched, neurologically normal individuals served as controls. In each instance, the autopsy was performed within 3 h after death. A statistically significant decrease in cell body area, number of synapses and total synaptic length was found in the normal-appearing neurons of the ALS patients. The alterations were more pronounced in neurons with central chromatolysis. However, despite an approximately 20  % reduction in the number of synapses, the length of the active synaptic zone of the normal-appearing neurons in the ALS patients was not diminished. This observation may be accounted for by a plasticity to the loss of synapses which maintained the active zone of the remaining synapses to increase synaptic efficiency. It is suggested that when the plasticity of the active zone reaches its limit, the continuing loss of synapses may lead to functional impairment. The capacity of the active synaptic zone to respond to progressive denervation of the anterior horn neurons may preserve motor function or slow the development of motor deficits in the early stage of degeneration of the lower motor neurons.

Notes: Abstract The spinal cord of 20 patients with amyotrophic lateral sclerosis (ALS) and 5 patients with lower motor neuron disease (LMND) were investigated immunohistochemically using anti-human excitatory amino acid transporter 1 (EAAT1) and EAAT2 antibodies which are the astrocytic transporters. The purpose of the study was to examine relationships between EAAT1 and EAAT2 immunoreactivity and degeneration of anterior horn neurons. Specimens from 20 patients without any neurological disease served as controls. In controls, spinal cord gray matter was densely immunostained by antibodies, whereas the white matter was generally not immunostained. In motor neuron disease (MND) patients, EAAT1 immunoreactivity was relatively well preserved in the gray matter despite neuronal loss of anterior horn cells. On the other hand, EAAT2 immunoreactivity in anterior horns correlated with the degree of neuronal loss of anterior horn cells: in the patients with mild neuronal depletion, anterior horns were densely immunostained by the antibody, whereas in the patients with severe neuronal loss, EAAT2 expression was markedly reduced. Degenerated anterior horn cells frequently showed a much denser EAAT1 and EAAT2 immunoreactivity around the surface of the neurons and their neuronal processes than that observed in normal-appearing neurons. There was no difference in the expression of EAAT1 and EAAT2 immunoreactivity between LMND and ALS patients. These findings suggest that in the early stage of degeneration of anterior horn cells, EAAT1 and EAAT2 immunoreactivity is preserved in the astrocytic foot directly attached to normal-appearing neurons, whereas levels of EAAT1 and EAAT2 protein rather increase in the astrocytic foot directly attached to degenerated anterior horn neurons; the latter effect most probably reduces the elevated glutamate level, compensates for the reduced function of astroglial glutamate transporters, or represents a condensation of EAAT1 and EAAT2 immunoreactivity secondary to loss of neurites and greater condensation of astrocytic processes. Thus, we demonstrate a difference in EAAT1 and EAAT2 immunoreactivity in different stages of progression in ALS, as a feature of the pathomechanism of this disease.

Notes: Summary Extracellular and intracellular recordings were made from spinocerebellar tract neurones of the central cervical nucleus (CCN) in C1–C3 segments of the anaesthetized cat. These neurones were identified by antidromic activation from the cerebellar peduncle. Stimulation of the ipsilateral dorsal root elicited extracellular spikes or EPSPs with a monosynaptic latency in almost all CCN neurones in the same segment (segmental input). Late excitatory effects were also observed in about one third of CCN neurones. The monosynaptic EPSP was occasionally followed by an IPSP. The excitatory input from the dorsal root to CCN neurones was extended over several segments for some CCN neurons (extrasegmental input). Monosynaptic excitation was evoked in CCN neurones after stimulation of dorsal neck muscle nerves as well; i.e. splenius (SPL), biventer cervicis and complexus (BCC), rectus capitus dorsalis, and obliquus capitus caudalis. Thresholds for this excitation were near the threshold of the nerve, suggesting that it originated from group I fibres. The component of excitation added after strong stimulation of neck muscle nerves would be attributed to group II fibres. When a CCN neurone received excitatory input from the nerve of one muscle, it was generally not affected by stimulation of other nerves in the same segment. Such muscle specificity of segmental input was the principal pattern of connexion of neck muscle afferents with CCN neurones. In some cases, however, excitatory convergence from SPL and BCC nerves onto single CCN neurones or excitation from the SPL nerve and inhibition from the BCC nerve were also observed. Nearly half of the CCN neurones received input from one muscle nerve of the same segment and not from the afferent of the same muscle of different segments, indicating a segment specificity of input. In the remaining CCN neurones, weaker excitatory effects were induced from afferents of different segments as well. In such extrasegmental effects, inputs to CCN neurones from caudal segments predominated in frequency over those from rostral segments. The origin of extrasegmental input was generally confined to the same muscle. Low threshold muscle afferents from the SPL and BCC were intraaxonally stained with HRP. The collaterals of the stained fibre distributed branchlets and terminals to the CCN, laminae VII, VIII, and motor nuclei. Two fibres responding to local muscle prodding or stretch showed a similar morphology. The findings indicated that muscle spindle afferents from primary endings projected to the CCN.

Notes: Summary Spinocerebellar tract (SCT) neurones in and around the central cervical nucleus (CCN) were physiologically identified by antidromic activation of these cells on stimulation of the cerebellum. Among the Spinocerebellar tract cells thus identified, those ascending the contralateral spinal funiculi were found in the CCN and ventralwards, whereas those ascending the ipsilateral funiculi existed mostly dorsal to the CCN partly overlapping with crossed cells in the nucleus. Mapping sites from which CCN cells were antidromically activated showed that axons of the CCN-SCT cross at the same segment, ascend the ventral funiculus initially, the lateral funiculus at rostral C1 and the lateral border of the medulla to reach the cerebellar peduncle, enter the cerebellum mainly via the restiform body but possibly also via the superior peduncle. Systematic mapping of stimulation within the cerebellum indicated that the CCNSCT projects to the medial part of the anterior lobe and the posterior lobe bilaterally. Projection to lobules I–II was found in almost all CCN-SCT cells examined. Three fourths of CCN-SCT cells projected to the posterior lobe, as revealed by less extensive mapping. Mapping of axonal regions of the same single CCN-SCT cells showed that they project multifocally in the cerebellum, where projection to lobules I–II was common and that to other areas varied with individual cells. Conduction velocites decreased within the cerebellum probably as the result of repeated branching. Mossy fibre responses evoked on stimulation of the C2 dorsal root in cats with the transected dorsal funiculi were shown to be mediated mostly via the CCN-SCT. Mapping the field potential showed that the response was by far the largest in lobules I–II. This suggested that the terminals provided by the CCN-SCT are the densest in these lobules.

Notes: Summary 1. Projections of reticulospinal neurones (RSNs) in the nucleus reticularis pontis caudalis (N.r.p.c.) to dorsal neck motoneurones supplying splenius (SPL, lateral head flexor) and biventer cervicis and complexus (BCC, head elevator) muscles were studied in the cat anaesthetized with pentobarbiturate or α-chloralose. 2. Threshold mapping for evoking antidromic spikes revealed that most of RSNs tested projecting down to brachial segments but not to lumbar segments (C-RSNs) gave off collaterals to the gray matter of the upper spinal cord in C2–C3 segments. 3. Spike triggered averaging showed that negative field potentials were evoked after firing of a single C-RSN (single fibre focal synaptic potentials, FSPs) in the region of C2–C3 where large antidromic field potentials from nerves supplying SPL or BCC muscles were evoked. The single fibre FSPs ranged between 1 and 10 μV in amplitude and had latencies between 0.7 and 1.2 ms from the onset of the triggering spike. In most cases, a presynaptic spike preceded the negative potential by 0.3 ms. These results indicated that C-RSNs project to the SPL or BCC motor nucleus. 4. Spike triggered averaging of postsynaptic potentials revealed EPSPs (single fibre EPSPs) in 36 dorsal neck motoneurones, predominantly in SPL (25) and less in BCC (11) motoneurones, evoked from 15 C-RSNs. The amplitude of the single fibre EPSPs ranged from 5 to 310 μV, and had latencies of 0.8–2.0 ms from the onset of the triggering spikes of C-RSNs, or 0.3–0.5 ms from the presynaptic spike when recorded. The results indicated monosynaptic excitatory connexions of C-RSNs to dorsal neck motoneurones. 5. Single fibre EPSPs from a C-RSN were usually recorded from either BCC or SPL motoneurones but not from both types of motoneurones, when tested in many motoneurones. This showed that connexions of C-RSNs with dorsal neck motoneurones were muscle specific. 6. RSNs projecting down to the lumbar segment (L-RSN) also showed branching in C2–C3 segments. Excitatory monosynaptic connexion of L-RSNs with neck motoneurones were demonstrated by recording single fibre postsynaptic population potentials (p.s.p.p.s.) from the C2 ventral root perfused with sucrose. The probability of evoking monosynaptic single fibre p.s.p.p.s. was less (19%) than for C-RSNs (59%).

Notes: Summary Dorsal neck motoneurones receive disynaptic tectal and pyramidal EPSPs via common reticulospinal neurones (RSNs). This study was aimed at identification of the RSNs projecting directly to neck motoneurones and mediating these EPSPs. 1. Stimulation of the tectum and the cerebral peduncle evoked monosynaptic descending volleys in the spinal cord, which were chiefly mediated by reticulospinal neurones in the pons and the medulla. Systematic tracking of the C3 and C7 segments was made to locate descending volleys in the spinal funiculi. The tectal monosynaptic volley was largest in the medial part of the ventral funiculus and decreased gradually as the recording electrode was moved to the lateral part of the ventral funiculus and the lateral funiculus. In contrast, the peduncle-evoked monosynaptic volley was distributed rather evenly in the ventral funiculus and the ventral half of the lateral funiculus. 2. Differences in funicular distribution of the two descending volleys suggest the existence of subgroups of RSNs which differed in strength of inputs from the two descending fibre systems and in the funicular location of descending axons. 3. The RSNs were classified into the following four groups; (1) mRSNs which descended in the medial part of the ventral funiculus, (2) in RSNs which descended in the ventrolateral funiculus, (3) 1RSNs which descended in the dorsal 2/3 of the lateral funiculus and (4) coRSNs which descended in the contralateral funiculi. The mRSNs were located in a fairly localized region corresponding to the nucleus reticularis pontis caudalis (N.r.p.c.), while inRSNs, 1RSNs and coRSNs were mainly in the nucleus reticularis gigantocellularis (N.r.g.), in the nucleus reticularis magnocellularis (N.r.m.) and in the nucleus reticularis ventralis (N.r.v.). RSNs were further divided into three types depending on the levels of projection. L-RSNs projected to the lumbar spinal segments. C-RSNs descended to the C6–C7 spinal segment but not to the lumbar segments. N-RSNs projected to the C3 but not to the C6–C7 segments. 4. Stimulation of the tectum and the cerebral peduncle produced monosynaptic negative field potentials in the medial two thirds of the reticular formation in the pons and medulla. Tectal field potentials were largest in the N.r.p.c. and the rostral part of the N.r.g., while pyramidal field potentials were largest in the N.r.g. Correspondingly, RSNs in the N.r.p.c. (mRSNs) received larger monosynaptic EPSPs from tectal than from pyramidal volleys, while RSNs in the N.r.g. (in-, 1- and coRSNs) received stronger input from the peduncle than from the tectum. 5. Stimulation of the C7 ventral but not the lateral funiculus evoked monosynaptic EPSPs on all the dorsal neck motoneurones tested. Stimulation of the L1 segment only produced monosynaptic EPSPs in 35% of the motoneurones. The L1 evoked EPSPs were much smaller than C7 evoked EPSPs. 6. The C7 evoked EPSPs (C7 EPSP) showed complete occlusion (collision) with the tectal or pyramidal disynaptic EPSPs. Similar results were obtained with L1 EPSPs. These results indicate that tectal and pyramidal disynaptic EPSPs in dorsal neck motoneurones were mediated chiefly by C-mRSNs and C-inRSNs and partly by L-RSNs.