Synaptic organization in the pacemaker nucleus of a medium-frequency weakly electric fish,Eigenmannia sp

The medullary command nucleus (MCN) of the medium-frequency weakly electric fish,Eigenmannia sp., contains two types of neurones, namely large and small cells, which are embedded in a neuropile of large and small myelinated fibers. Using serial semi-thin and ultra-thin sectioning, combined with HRP labelling established that both cell types possess rich dendritic arborization and large myelinated axons. Only the axons of the large cells leave the nucleus and these contribute the unique output of the MCN. Axon branching has been observed only in the axons of small cells and their collaterals show an exclusively intranuclear course. Two types of synaptic terminals have been found on large as well as on small cells: (1) large club endings forming both gap (electronic) junctions and polarized chemical synapses, which often appear at the same junction constituting morphologically mixed synapses; and (2) small bouton-like terminals forming exclusively chemical synaptic contacts. No differences between the two neuron types could be detected with respect to the arrangement of the synaptic contacts: club endings and small bouton-like terminals synapse on dendritic processes as well as on perikarya, while the unmyelinated initial segments were always found to be free of synaptic contacts. Large and small cells were found to be simultaneously connected by the same club ending or small bouton-like terminal: in the case of club endings by means of gap junctions and chemical synapses, whereas in the case of boutons by chemical synapses only. Club endings sometimes form gap junctions with each other. The possible role of these unusual synaptic connections in local synchronization is suggested. Club endings originate from the large axons of small cells, while small bouton-like terminals originate from the fine myelinated fibers of extranuclear origin. InEigenmannia, small cells, being connected to large cells as well as to each other by axo-somatic and axodendritic synapses, can be considered as the pacemaker cells of the MCN whereas large cells are relay cells. Small bouton-like terminals may convey exogeneous impulses towards the MCN exerting modulatory effects at both pacemaker and relay cell levels. The greater variety of ultrastructural correlates established in the MCN ofEigenmannia, in comparison withSternarchus⁵ (see also ref. 16), suggests increased modulation possibilities in the former fish's EOD behaviour.     

The electromotor system of the electric eel investigated with horseradish peroxidase as a retrograde tracer

The electromotor system of the electric eel, Electrophorus electricus, was studied by injection of horseradish peroxidase as a retrograde tracer. The electromotor neurons, which innervate the electrocytes, comprise a midline nucleus, largely dorsal to the spinal canal. Spinal motoneurons lie ventrolaterally. The electromotor and skeletal motor neuron populations correspond to the acetylcholinesterase-negative and -positive cells previously described. The medullary relay neurons were labeled following HRP injection into the spinal cord at a level where electromotor neurons occurred, but not after injection into the cord in the abdominal region rostral to these cells. Other medullary neurons, presumably bulbospinal motor fibers, were labeled after both levels of spinal cord injection. The results suggest that these axosomatic synapses, which are electrically transmitting but morphologically mixed, take up retrograde tracers in a manner similar to chemical synapses and that tracer uptake is at least largely at terminal regions.     

Parvocells: a novel interneuron type in the pacemaker nucleus of a weakly electric fish

Gymnotiform weakly electric fish produce electric organ discharges (EODs) that function in electrolocation and communication. The command signal for the EOD is produced by the medullary pacemaker nucleus, which contains two well-characterized neuron types: pacemaker cells and relay cells. In this study, we characterized a third neuron type in the pacemaker nucleus. These neurons, which we have named parvocells, were smaller (7-15 microm in diameter) than relay and pacemaker cells. The parvocells were labeled with an antibody against the neuronal calcium-binding protein, parvalbumin, and were not labeled with several glial-specific antibodies. Parvocells had one to three fine processes that often terminated at the periphery of relay and pacemaker cell bodies. The parvalbumin-positive terminals of the parvocells colocalized with immunoreactivity for SV-2, suggesting that the parvocells form chemical synapses on the relay and pacemaker cells. Parvalbumin-positive neurons are frequently gamma-aminobutyric acid (GABA)ergic or glycinergic, and the cytoplasm of the parvocell somata was immunoreactive with a glycine antibody. Antibodies against glycine receptors and gephyrin, however, did not label any cells in the pacemaker nucleus, suggesting that the pacemaker nucleus does not contain glycine or GABA((A)) receptors. Electron microscopy revealed gap junctions between the membranes of parvocells and adjacent terminal-like structures. Furthermore, neurobiotin injected into individual pacemaker or relay cells labeled parvocells as well as other pacemaker and relay cells, demonstrating that the parvocells are dye-coupled to the other neuron types in the pacemaker nucleus. These findings indicate that the parvocells are histochemically distinct from relay and pacemaker cells and that they receive electrotonic inputs from and make chemical synapses back onto pacemaker and relay cells. Further study is needed to investigate the function of these neurons in regulating the EOD.     

Morphology and physiology of the brainstem nuclei controlling the electric organ discharge in mormyrid fish

The motoneurons to the mormyrid electric organ are driven from the medullary relay nucleus. This nucleus is in turn innervated by an adjacent cell group, nucleus C. The goals of this study were to characterize the morphology and physiology of neurons in these two nuclei, and to test the hypothesis that nucleus C is the command nucleus responsible for initiating the electric organ discharge. Medullary relay neurons and nucleus C neurons were recorded intracellularly and labeled with horseradish peroxidase. Medullary relay neurons have a richly branched dendritic arborization, confined mainly to the nucleus itself, and somatosomatic, dendrosomatic, and presynaptic dendro-axonal gap junctions have been observed. Medullary relay neuron axons descend to the spinal cord without branching. Nucleus C dendrites extend far into the ventral reticular formation. Axons of nucleus C neurons have one branch that ramifies densely within the medullary relay nucleus, forming large club endings on the medullary relay neuron soma. Two additional branches project bilaterally toward the bulbar command associated nuclei. Both medullary relay neurons and nucleus C neurons fire a double action potential that precedes each electric organ discharge. Activity in nucleus C precedes that in the medullary relay nucleus by 100-300 microseconds. Postsynaptic activity is recorded in nucleus C neurons but not in medullary relay neurons. Hyperpolarization of a single nucleus C neuron can lower the frequency of the electric organ discharge. Both morphological and physiological data indicate that nucleus C is an integrating center where the electric organ command is initiated.     

Calretinin-like immunoreactivity in mormyrid and gymnarchid electrosensory and electromotor systems

Calretinin-like immunoreactivity was examined in the electrosensory and electromotor systems of the two families of mormyriform electric fish. Mormyrid fish showed the strongest immunoreactivity in the knollenorgan electroreceptor pathway; in the nucleus of the electrosensory lateral line lobe (ELL) and the big cells of the nucleus exterolateralis pars anterior. Mormyromast and ampullary zones of the ELL showed calretinin-like immunoreactivity in the ganglion, granule, and intermediate cell and fiber layers. Mormyromast zones additionally showed labeling of apical dendrites and commissural cells, but the ampullary zone did not. In the electromotor system, two nuclei in the corollary discharge pathway showed labeling: in the paratrigeminal command-associated nucleus and the juxtalobar nucleus. Gymnarchus niloticus (Gymnarchidae) showed strongest calretinin-like immunoreactivity in part of the phase-coding pathway; in S-type electroreceptor afferents. Zones of the ELL not receiving phase-coder input had weak labeling. The electromotor system showed labeling in the lateral relay nucleus and less strongly in the medullary relay nucleus, but none in the pacemaker. The concentration of calcium-binding proteins in mormyrid and gymnarchid time-coding electrosensory pathways is consistent with the hypothesis that they play a role in preserving temporal information across synapses. Cell types that encode temporal characteristics of stimuli in precise spike times have high levels of calcium-binding proteins, but cells that re-code temporal information into presence or magnitude of activity have low levels. Some cell types in the electromotor pathways and early in the time-coding electrosensory pathways do not follow this hypothesis, and therefore preserve temporal information using a mechanism independent of calcium-binding proteins. In particular, electromotor systems may use extensive electrotonic coupling within nuclei to ensure precise timing. J. Comp. Neurol. 387:341–357, 1997. © 1997 Wiley-Liss, Inc.     

Electron microscopic immunocytochemical study of the distribution of parvalbumin-containing neurons and axon terminals in the primate dentate Gyrus and Ammon's horn

Five green monkeys were examined with light and electron microscopic preparations to explore the regional differences in the distribution of parvalbumin (PV)-positive neurons and axon terminals in the primate hippocampus. PV-positive neurons were mainly found in the hilus of the dentate gyrus and the strata oriens and pyramidale of Ammon's horn. In electron microscopic preparations, the PV-positive cells displayed nuclear infoldings, intranuclear rods, a large rim of perikaryal cytoplasm with numerous organelles and both asymmetric and symmetric axosomatic synapses. One prominent PV-positive cell type in CA1 was a large multipolar neuron that resembled the large basket cells of the neocortex. Although most PV-positive dendrites were aspiny and postsynaptic to numerous axon terminals, some PV-positive dendrites in the molecular layer of the dentate gyrus displayed filipodia-like appendages with no synapses or spines that were postsynaptic to multiple axon terminals. The PV-positive dendrites in the hilus and stratum oriens were apposed at specialized junctions that resembled gap junctions. PV-positive axons were concentrated in the principal cell layers, and formed axosomatic, axodendritic, and axon initial segment synapses. In cases where these axons were observed to appose the surface of granule cells for a long length, only one axosomatic symmetric synapse per cell was found. In the hilus, PV-positive axon terminals formed synapses onto thorny excrescences of spiny cells. Both semithin sections and electron microscopic preparations indicated that more PV-positive axon terminals formed symmetric axosomatic synapses with pyramidal cells in CA2 than in CA1 and CA3. Also, CA2 displayed a unique plexus of PV-positive axon terminals in stratum lacunosum moleculare. These results indicate that the PV-positive hippocampal cells form a subset of GABAergic local circuit neurons, including the basket and chandelier cells. The ubiquitous finding of PV-positive dendrites linked by gap junctions throughout the dentate gyrus and Ammon's horn adds further data to indicate that this subset of GABAergic neurons is linked electrotonically. The synaptic organization of PV-positive neurons in the hippocampus suggests their participation in both feedback and feedforward inhibition. The PV-positive neurons in the hippocampus are only a proportion of the basket and chandelier cells, whereas virtually all of these cells in neocortex are PV-positive. © 1993 Wiley-Liss, Inc.     

Synaptic distribution of afferents from reticular nucleus in ventroposterior nucleus of cat thalamus

This study was aimed at determining the synaptic circuitry that contributes to the alterations in thalamic function that accompany changes in behavioral states. The somatosensory sector of the thalamic reticular nucleus (RTN) was identified by microelectrode recording in cats and injected with Phaseolus vulgaris-leucoagglutinin (PHA-L). The axons of labeled RTN cells gave rise to collaterals within the RTN and continued into the dorsal thalamus where they terminated predominately in the ventral posterior lateral nucleus (VPL). After small injections in the upper limb representation of RTN, most labeled terminations in VPL were confined to its medial part, suggesting the presence of a topographic organization in the projection. Terminations were concentrated in localized, focal aggregations of boutons. Combined electron microscopic immunocytochemistry, using immunogold labeling for γ-aminobutyric acid (GABA), showed that the PHA-L labeled boutons were GABA-positive terminals that ended in symmetrical synapses. Eighty-two percent of these synapses were on dendrites of relay neurons, 8.5% on dendrites of interneurons, and 9.3% on somata. The terminals of RTN axons form the majority of axon terminals ending in symmetrical synapses in VPL. Their concentration on relay neurons probably underlies the capacity of the RTN projection to reduce background activity of VPL relay neurons in the awake state and to maintain oscillatory behavior of these neurons in drowsiness and early phases of Sleep. © 1995 Wiley-Liss, Inc.     

Ultrastructural correlates of electrical-chemical synaptic transmission in goldfish cranial motor nuclei

The output connections of the cranial relay neurons, part of the Mauthner cell network, were examined in goldfish with light and electron microscopic techniques. Either lucifer yellow or horseradish peroxidase (HRP) was injected into cranial relay neuron axons to demonstrate that they diverge to several motor nuclei and to many motoneurons within one nucleus. Retrograde transport of the enzyme from injections of mandibular muscles was used to label the trigeminal motoneurons. In the electron microscope, cranial relay neuron processes were distinguished by the granular appearance of the electron-opaque polymer formed enzymatically by HRP, while the retrogradely labeled motoneurons had the polymer enclosed in lysosomes. The cranial relay neuron terminals contained many presynaptic vesicles which concentrated the HRP reaction product. Active zones and synaptic clefts were evident. At some synapses, both gap junctions and presynaptic vesicles were found. The mechanism of synaptic transmission was investigated by simultaneous recording with two intracellular microelectrodes from cranial relay neuron-motoneuron pairs. Composite postsynaptic potentials in a trigeminal motoneuron were evoked by intracellular stimulation of a cranial relay neuron axon. The earliest excitatory postsynaptic potential (EPSP) component had a latency of 0.25 msec and had a peak amplitude that was not depressed by repetitive stimulation. A second component had larger peak amplitudes which were reduced easily by repetitive stimulation. Antidromic action potentials were not transmitted from motoneurons to the cranial relay neuron axons. Thus, both electrical and chemical transmission probably occur at the cranial relay neuron-motoneuron synapses. Since the cranial relay neurons fire synchronously and receive excitatory chemical synapses, the function of the gap junctions and electrical transmission is unclear. Perhaps the importance of these gap junctions is more for transport of small molecules than for impulse transmission.     

Neuronal gap junctions and morphologically mixed synapses in the spinal cord of a teleost, Sternarchus albifrons (gymnotoidei)

Spinal cord motoneurons in the gymnotid, Sternarchus albifrons, were studied electron microscopically with special reference to the freeze-fracture method. Two types of motoneurons were identified. Electromotor neurons are monopolar and are located in a midline column dorsal to the ventral gray. These cells have a small fraction of their surface covered by synapses from descending axons, often at nodes. The synapses have multiple gap junctions, but few presynaptic vesicles or other correlates of chemical transmission. The gap junctions have an ordinary appearance in freeze-fracture replicas and exhibit a highly ordered substructure. The not infrequent appositions between the cell bodies of electromotor neurons exhibit no junctional specializations. Ordinary motoneurons are multipolar and densely covered with axosomatic and axodendritic synapses. In thin sections these synapses can be divided into two groups according to whether the vesicles are spherical or flattened. Gap junctions occur only at the first type, thus forming ‘morphologically mixed’ synapses. In freeze-etch replicas of motoneurons, the gap junctions are often found near clusters of postsynaptic E face particles elsewhere associated with excitatory chemical transmission. In addition, vesicle attachment sites occur in the presynaptic membranes of some synapses with gap junctions. The morphological observations are consistent with dual chemical and electrical transmission at these particular synapses, i.e. electrical excitation across gap junctions and chemical excitation at active zones with spherical vesicles and post-synaptic E face particles.     

Morphology and synaptic connections of myelinated primary axons in the ventrolateral region of rat trigeminal nucleus oralis

Neurons in the ventrolateral (VL) subdivision of rat trigeminal nucleus oralis (Vo) have most of their dendritic arbors confined within this region. This study examines the morphology and synaptic connections of a population of myelinated primary trigeminal axons that arborize within VL and are in a position to provide input directly to VL neurons. Primary axons were visualized for light and electron microscopic analysis by injecting 30% horseradish peroxidase (HRP) in 2% dimethylsulfoxide (DMSO) into the sensory root of the trigeminal nerve and allowing 24-36 hours for the anterograde transport of HRP into the terminal axonal arbors. This population is characterized by its cone-shaped terminal arbors, which generate many axonal endings (2-8 micron in diameter) along unmyelinated terminal strands. These arbors arise from collaterals emanating from thinly myelinated (2-5 micron in diameter) parent branches descending in the spinal V tract, which, on the basis of their size, are considered to be small myelinated (A sigma) primary trigeminal axons. HRP-labeled P endings belonging to this population of primary axons are scalloped, filled with spherical to ovoid (40-70 nm in diameter) synaptic vesicles, and lie centrally in glomeruli where they make asymmetrical axodendritic synapses on dendritic shafts and spine heads. It is at these synapses that this population of primary trigeminal axons is probably transferring its input directly to the dendritic arbors of VL neurons. The dendritic shafts and spine heads also receive symmetrical to intermediate axodendritic synapses from endings containing flattened (70 X 29 nm) synaptic vesicles. These terminals also establish axo-axonic synapses on the P ending. Other synaptic components found less often in the glomeruli include small terminals containing oval (14-23 nm) synaptic vesicles that establish symmetrical to intermediate synapses on the P ending, boutons containing pleomorphic (35-80 nm) synaptic vesicles that form symmetrical to intermediate synapses on the P ending as well as on dendritic shafts, and small peripheral endings containing round (20-40 nm) synaptic vesicles that establish asymmetrical synapses on dendritic shafts.     

Predominance of corticothalamic synaptic inputs to thalamic reticular nucleus neurons in the rat.

Quantitative electron microscopy was used to examine the relative contributions of different types of synapses to the circuitry of the thalamic reticular nucleus (RTN) in the rat. Single RTN cells were injected with Lucifer Yellow (LY) in fixed brain slices and examined after photoconversion; corticothalamic axons and terminals were labeled by anterograde transport of Phaseolus vulgaris-leucoagglutinin (PHA-L); and gamma-aminobutyric acid (GABA)ergic terminals were labeled by postembedding immunocytochemistry. Three types of synapses, made by morphologically distinguishable small terminals (ST), large terminals (LT), and GABAergic terminals, were distributed on all portions of the dendritic trees of injected RTN cells. ST and LT terminals formed asymmetrical, presumed excitatory, synaptic contacts. On proximal dendrites, approximately 50% of the synapses were ST, 30-40% were LT, and 10-25% were GABAergic. On distal dendrites, 60-65% were ST, 20% were LT, and 15% were GABAergic. PHA-L labeling showed that labeled corticothalamic terminals and ST terminals have identical morphological features and the same distribution patterns on RTN dendrites, indicating that the majority of excitatory afferents to RTN neurons are derived from the cerebral cortex. The LT terminals found in smaller numbers are probably derived from collateral axons of thalamocortical relay cells. GABAergic terminals formed by LY-labeled, intra-RTN axon collaterals were relatively few in number, and no dendrodendritic synapses were observed.     

The midbrain precommand nucleus of the mormyrid electromotor network.

The functional role of the midbrain precommand nucleus (PCN) of the electromotor system was explored in the weakly electric mormyrid fish Gnathonemus petersii, using extracellular recording of field potentials, single unit activity, and microstimulation in vivo. Electromotor-related field potentials in PCN are linked in a one-to-one manner and with a fixed time relationship to the electric organ discharge (EOD) command cycle, but occur later than EOD command activity in the medulla. It is suggested that PCN electromotor-related field potentials arise from two sources: (1) antidromically, by backpropagation across electrotonic synapses between PCN axons and command nucleus neurons, and (2) as corollary discharge-driven feedback arriving from the command nucleus indirectly, via multisynaptic pathways. PCN neurons can be activated by electrosensory input, but this does not necessarily activate the whole motor command chain. Microstimulation of PCN modulates the endogenous pattern of electromotor command in a way that can mimic the structure of certain stereotyped behavioral patterns. PCN activity is regulated, and to a certain extent synchronized, by corollary discharge feedback inhibition. However, PCN does not generally function as a synchronized pacemaker driving the electromotor command chain. We propose that PCN neurons integrate information of various origins and individually relay this to the command nucleus in the medulla. Some may also have intrinsic, although normally nonsynchronized, pacemaker properties. This descending activity, integrated in the electromotor command nucleus, will play an important modulatory role in the central pattern generator decision process.     

Morphology and synaptic connections of ultrafine primary axons in lamina I of the spinal dorsal horn: candidates for the terminal axonal arbors of primary neurons with unmyelinated (C) axons

Neurons in Rexed's lamina I have the bulk of their dendritic arbors confined within this lamina. This study examines the morphology and synaptic connections of primary axons which generate axonal endings in lamina I of the spinal dorsal horn and are in position to deliver their inputs directly to lamina I neurons. Primary axons were made visible for light and electron microscopical study by applying horseradish peroxidase (HRP) to the severed central stumps of cervical and lumbar dorsal roots and allowing sufficient time for the orthograde movement of the HRP into the terminal axonal arbors. Golgi preparations provided supplementary light microscopical views of these axons. Lamina I receives the terminal arborization of two very different kinds of primary axons. One of these generates many ultrafine endings along unbranched, long rostrocaudally oriented, strand-like collaterals which arise from thin parent branches in Lissauer's tract. In view of these thin parent branches, most ultrafine primary axons are considered to be unmyelinated (C) primary axons. The second kind of primary axon generates large caliber endings on branched collaterals. These arise from relatively thick parent branches in Lissauer's tract which, on the basis of their size, are considered to be myelinated (A delta) primary axons. The scalloped endings of both primary axons lie in the interior of glomeruli where they form axodendritic synapses on small dendritic shafts and spines. It is at these synapses that these two kinds of primary axons are thought to transfer nociceptive and thermal inputs directly to the dendritic arbors of lamina I neurons. Transmitter release at these axodendritic synapses in response to primary inputs can be modified, probably diminished or inhibited, by synaptic events within the glomeruli from at least three sources. Synaptic vesicle-containing dendrites form dendroaxonic synapses on primary endings and two kinds of axons form axoaxonic synapses either on primary endings or on the intervaricose segments of the primary axons.     

Termination in trigeminal nucleus oralis of ascending intratrigeminal axons originating from neurons in the medullary dorsal horn: an HRP study in the rat employing light and electron microscopy

The anterograde horseradish peroxidase (HRP) technique was used to identify ascending intratrigeminal axons originating from neurons in the medullary dorsal horn (MDH) which terminate in trigeminal nucleus oralis (Vo). HRP injections into the MDH labeled two populations of axons ascending ipsilaterally within the spinal trigeminal nucleus. The first population was composed of parent branches which each gave off a single branching collateral strand to Vo as they ascended. These collaterals were characterized by boutons filled with small, round synaptic vesicles and forming asymmetrical synaptic contacts with large diameter dendritic shafts. The second axonal population was made up of parent branches which terminated directly in Vo. Their short terminal strands were distinguished by axonal endings containing pleomorphic synaptic vesicles and forming symmetrical synaptic junctions with small diameter dendritic shafts and spines.     

Reorganization of corticorubral synapses following cross-innervation of flexor and extensor nerves of adult cat: a quantitative electron microscopic study

A quantitative electron microscopic study of corticorubral synapses was performed in the red nucleus (RN) of adult cats to determine the morphological correlates for the changes in time course of corticorubral excitatory post-synaptic potentials, which occur following cross-innervation of forelimb extensor and flexor nerves. Corticorubral synaptic endings were identified by anterograde degeneration after lesions of the ipsilateral sensorimotor cortex. Rubrospinal neurons innervating upper spinal segments were electrophysiologically identified and filled with horseradish peroxidase (HRP). These cells were mainly situated in the dorsomedial part of RN. Electron micrographs of the degenerating corticorubral synaptic endings were taken in the region surrounding HRP-filled neurons and the diameter of the dendrites contacted by such terminals was measured.In the cross-innervated animals many degenerating terminals were found to synapse on dendrites with large diameter and the somata of neurons in RN. This is in contrast to the previous observations in normal cats, in which very few corticorubral synapses were found to synapse on proximal dendrites and somata of RN neurons. The diameter of HRP-filled neurons in cats which were cross-innervated was slightly smaller than those observed in normal animals. These results indicate that new corticorubral synapses were formed on proximal dendrites and somata of RN neurons as a consequence of cross-innervation.     

Synaptic organization of individual neurons in the macaque lateral geniculate nucleus

Parvocellular and magnocellular neurons in the dorsal lateral geniculate nucleus of macaque monkeys were recorded electrophysiologically and then injected with HRP. The injected neurons were examined with the electron microscope. Synaptic terminals contacting the dendrites of individual neurons were classified and the synapses counted to estimate the number and distribution of each type over the entire dendritic tree. Seven parvocellular and 2 magnocellular neurons were analyzed. Two of the parvocellular neurons had presynaptic dendrites and no axons. These interneurons had electro-physiological characteristics much like those of relay neurons with the exception that their receptive field center responses had the opposite sign; i.e., they had OFF centers, while most neurons around them had ON centers. All of the relay neurons had similar types and distributions of terminal contacts. However, the distribution of each synaptic type along the dendrites of an individual neuron was not homogeneous. Retinal and F terminals were located predominantly on proximal dendrites whereas RSD terminals, either from the cortex and/or brain stem, predominated on the intermediate and distal dendrites. Parvocellular neurons were estimated to have about 500 total synapses on their dendritic trees, while magnocellular neurons had about 3000 total synapses on their dendritic trees. The retinal terminals making synaptic contacts with magnocellular neurons were also presynaptic to terminals containing flattened vesicles; these latter terminals also had synapses onto the magnocellular neuron's dendrites. Such a synaptic arrangement is called a triadic arrangement, or triad. Parvocellular neurons rarely had such triadic arrangements. In comparing these data with those of the cat, it was concluded that the major synaptic difference between relay cell types in both species (Class 1/Class 2 cells for the cat and parvo/magno cells for the monkey) was the frequent occurrence of triads for Class 2 cells and magnocellular cells versus the infrequent occurrence of triads for Class 1 cells and parvocellular cells. Although these triadic arrangements have been studied for over 2 decades, their function has yet to be determined, but probably relates to inhibition of retina signals at dendrites of magnocellular neurons in the monkey and Class 2 cells in the cat.     

Dual ultrastructural immunocytochemical labeling of μ and δ opioid receptors in the superficial layers of the rat cervical spinal cord

The δ opioid receptor (DOR) and μ opioid receptor (MOR) are abundantly distributed in the dorsal horn of the spinal cord. Simultaneous activation of each receptor by selective opiate agonists has been shown to result in synergistic analgesic effects. To determine the cellular basis for these functional associations, we examined the electron microscopic immunocytochemical localization of DOR and MOR in single sections through the superficial layers of the dorsal horn in the adult rat spinal cord (C2–C4). From a total of 270 DOR-labeled profiles, 49% were soma and dendrites, 46% were axon terminals and small unmyelinated axons, and 5% were glial processes. 6% of the DOR-labeled soma and dendrites, and <1% of the glial processes also showed MOR-like immunoreactivity (MOR-LI). Of 339 MOR-labeled profiles, 87% were axon terminals and small unmyelinated axons, 12% were soma and dendrites, and 2% were glial processes. 21% of the MOR-labeled soma and dendrites, but none of the axon terminals also contain DOR-LI. The subcellular distributions of MOR and DOR were distinct in axon terminals. In axon terminals, both DOR-LI and MOR-LI were detected along the plasmalemma, but only DOR-LI was associated with large dense core vesicles. DOR-labeled terminals formed synapses with dendrites containing MOR and conversely, MOR-labeled terminals formed synapses with DOR-labeled dendrites. These results suggest that the synergistic actions of selective MOR- and DOR-agonists may be attributed to dual modulation of the same or synaptically linked neurons in the superficial layers of the spinal cord.     

Synaptic displacement in intracentral neurons of Clarke's nucleus following axotomy in the cat

The dorsal spinocerebellar tract was severed unilaterally in 12 cats which were killed at 4, 15, 35, and 90 days. Clarke's neurons of L3 segment were studied by electron microscopy for chromatolytic, synaptic, and glial changes. Some large neurons of Clarke's nucleus were chromatolyzed by 4 days, and all large cells by 15 days. Small cells of Clarke's nucleus remained unchanged. At 35 and 90 days chromatolytic cells were all reduced in size. These atrophic cells were characterized by pale Nissl bodies, dark dendrites, synapses with pale matrix, and giant synaptic terminals. Synaptic changes were noted on chromatolyzed neurons by 4 days. The first signs of synaptic changes were reductions in the number of synaptic complexes, widening of synaptic clefts, and the appearance of cisternae under postsynaptic membrane of giant synapses. Occasional boutons with pale matrix and decreased numbers of synaptic vesicles were noted. There was a moderate increase in the astrocytic processes on the neuronal surface membrane. Some of these processes were inserted into the widened synaptic clefts. Some synapses remained unchanged. At 15 days the synaptic changes were similar but now occurred on all large cells. The synaptic matrix was less dense and the number of vesicles was less in the separated boutons. At 90 days all boutons, except giant synapses on the base of proximal dendrites, were small and some were arranged in dense patches. Some synapses were pale, and others were unchanged. The patchy distribution of numerous small boutons on the atrophic Clarke's neurons is considered to be due to collateral sprouting and formations of new terminals. These findings have implications in relation to spinal shock and recovery from it.     

Transneuronal transport of intracellularly injected biotinamide in primary afferent axons

Transneuronal transport of biotinamide was observed following intracellular injection of biotinamide into rat jaw-muscle spindle afferent axons. Microelectrodes were advanced into the mesencephalic nucleus of the trigeminal nerve where jaw-muscle spindle afferent axons were identified by their increased firing during stretching of the jaw-elevator muscles. Biotinamide (Neurobiotin) was then injected into individual axons and the animals were maintained under anesthesia for 2−6 h. The animals were then killed via an overdose of anesthetic and the brainstem was processed histochemically. Biotinamide-filled axon collaterals and terminals were readily visible in the trigeminal motor nucleus, the trigeminal sensory nuclei, and adjacent reticular formation. In addition to these intracellularly stained axons, two to five neurons per animal (total of 36 in eight rats) were observed with a homogenous gray reaction product distributed throughout their somata, proximal, and secondary dendrites. These neurons ranged in size from small (8–20 μm, n = 26) to medium-sized (<30 μm, n = 10) and were closely apposed by numerous (up to 20) biotinamide-stained spindle afferent boutons. Most of these neurons (n = 22) were located in the dorsomedial portion of the spinal trigeminal subnucleus interpolaris (Vi) 2.5−4.5 mm caudal to the intra-axonal injection site. Electron microscopic analysis in two rats suggests that the transneuronal biotinamide labeling occurred predominately through asymmetric, axodendritic synapses between biotinamide-filled axon terminals and Vi neuronal dendrites. Although recent in vitro studies have reported that biotinamide permeates through gap junctions, in this study we found no evidence of biotinamide traversing the gap junctions which exist between trigeminal mesencephalic nucleus (Vme) neuronal somata. These results demonstrate that biotinamide can occasionally be transneuronally transported presumably via synapses; further information is needed to explain the seemingly sporadic nature of this transport.     

An electron microscopic study of synaptic organization in the medial superior olive of normal and experimental chinchillas

The medial superior olive (MSO) was studied in normal animals to determine the types of synaptic endings and their distribution over the surface of MSO neurons. Unilateral lesions were made in the anteroventral cochlear nucleus (AVCN) of experimental animals to determine the source of at least one synaptic type in the MSO. The surfaces of MSO neurons in normal animals were studded with three distinct types of synaptic endings distinguished mainly by the size of their synaptic vesicles. There were endings with large vesicles, 510 Å in mean diameter; endings with small vesicles, 380 Å; and endings with vesicles intermediate in size. 435 Å. The large vesicle ending typically was greater than 2 μm in maximum diameter. It appeared as the termination of a myelinated axon or as a swollen portion of a node and made multiple asymmetrical synapses. Large vesicle endings occurred exclusively on dendrites where they formed 85% of the synaptic endings. Small vesicle endings typically were less than 2 μm in diameter. They appeared as the termination of a fine unmyelinated axon and made only one symmetrical synapse. Small vesicle boutons occurred infrequently over the entire neuronal surface. Intermediate vesicle synaptic endings were similar to large vesicle endings except that they were present only on the cell body, axon hillock, and proximal portions of the dendrites where they formed most of the synapses. In AVCN lesioned animals degenerating myelinated axons and large vesicle synaptic endings were distributed to the lateral dendrites of the ipsilateral MSO and medial dendrites of the contralateral one. In addition, a few degenerating axons and large vesicle endings were found among the ipsilateral medial dendrites. The changes in the degenerating endings were characterized by an early proliferation of neurofilaments and swelling of the endings followed by collapse of the endings and increase in electron density, disappearance of filaments and synaptic vesicles, and phagocytosis of the degenerated endings by reactive glial cells. No degenerative changes were observed in the small and intermediate vesicle endings. The results of this study indicate that the more numerous large vesicle endings presynaptic to the MSO dendrites are the axon terminals of neurons in the AVCN. The persistence after lesions of the small and intermediate vesicle endings suggests that they arise from as yet unidentified sources.