Vesicle-containing Dendrites Research Articles

Neurokinin-1 Receptor Immunoreactive Neuronal Elements in the Superficial Dorsal Horn of the Chicken Spinal Cord: With Special Reference to Their Relationship with the Tachykinin-containing Central Axon Terminals in Synaptic Glomeruli

Synaptic glomeruli that involve tachykinin-containing primary afferent central terminals are numerous in lamina II of the chicken spinal cord. Therefore, a certain amount of noxious information is likely to be modulated in these structures in chickens. In this study, we used immunohistochemistry with confocal and electron microscopy to investigate whether neurokinin-1 receptor (NK-1R)-expressing neuronal elements are in contact with the central primary afferent terminals in synaptic glomeruli of the chicken spinal cord. We also investigated which neuronal elements (axon terminals, dendrites, cell bodies) and which neurons in the spinal cord possess NK-1R, and are possibly influenced by tachykinin in the glomeruli. By confocal microscopy, NK-1R immunoreactivities were seen in a variety of neuronal cell bodies, their dendrites and smaller fibers of unknown origin. Some of the NK-1R immunoreactive profiles also expressed GABA immunoreactivities. A close association was observed between the NK-1R-immunoreactive neurons and tachykinin-immunoreactive axonal varicosities. By electron microscopy, NK-1R immunoreactivity was seen in cell bodies, conventional dendrites and vesicle-containing dendrites in laminae I and II. Among these elements, dendrites and vesicle-containing dendrites made contact with tachykinin-containing central terminals in the synaptic glomeruli. These results indicate that tachykinin-containing central terminals in the chicken spinal cord can modulate second-order neuronal elements in the synaptic glomeruli.

BDNF‐mediated modulation of GABA and glycine release in dorsal horn lamina II from postnatal rats

Recent studies show that excitatory glutamatergic transmission is potentiated by BDNF in superficial dorsal horn, both at the pre- and the postsynaptic site. The role of BDNF in modulating GABA and glycine-mediated inhibitory transmission has not been fully investigated. To determine whether the neurotrophin is effective in regulating the spontaneous release of the two neurotransmitters, we have recorded miniature inhibitory postsynaptic currents (mIPSCs) in lamina II of post-natal rats. We show that application of BDNF enhanced the spontaneous release of GABA and glycine, in presence of tetrodotoxin. The effect was blocked by the trk-receptor inhibitor k-252a. Amplitude and kinetics of mIPSCs were not altered. Evoked GABA and glycine IPSCs (eIPSCs) were depressed by BDNF and the coefficient of variation of eIPSC amplitude was significantly increased. By recording glycine eIPSCs with the paired-pulse protocol, an increase of paired-pulse ratio during BDNF application was observed. We performed parallel ultrastructural studies to unveil the circuitry involved in the effects of BDNF. These studies show that synaptic interactions between full length functional trkB receptors and GABA-containing profiles only occur at non peptidergic synaptic glomeruli of types I and II. Expression of trkB in presynaptic vesicle-containing dendrites originating from GABAergic islet cells, indicates these profiles as key structures in the modulation of inhibitory neurotransmission by the neurotrophin. Our results thus describe a yet uncharacterized effect of BDNF in lamina II, giving further strength to the notion that the neurotrophin plays an important role in pain neurotransmission.

GABAB receptors in the centromedian/parafascicular thalamic nuclear complex: An ultrastructural analysis of GABABR1 and GABABR2 in the monkey thalamus

Strong gamma-aminobutyric acid type B (GABA(B)) receptor binding has been shown throughout the thalamus, but the distribution of the two GABA(B) receptor subunits, GABA(B) receptor subunit 1 (GABA(B)R1) and GABA(B) receptor subunit 2 (GABA(B)R2), remains poorly characterized. In primates, the caudal intralaminar nuclei, centromedian and parafascicular (CM/PF), are an integral part of basal ganglia circuits and a main source of inputs to the striatum. In this study, we analyzed the subcellular and subsynaptic distribution of GABA(B) receptor subunits by using light and electron microscopic immunocytochemical techniques. Quantitative immunoperoxidase and immunogold analysis showed that both subunits display a similar pattern of distribution in CM/PF, being expressed largely at extrasynaptic and perisynaptic sites in neuronal cell bodies, dendrites, and axon-like processes and less abundantly in axon terminals. Postsynaptic GABA(B)R1 labeling was found mostly on the plasma membrane (70-80%), whereas GABA(B)R2 was more evenly distributed between the plasma membrane and intracellular compartments of CM/PF neurons. A few axon terminals forming symmetric and asymmetric synapses were also labeled for GABA(B)R1 and GABA(B)R2, but the bulk of presynaptic labeling was expressed in small axon-like processes. About 20% of presynaptic vesicle-containing dendrites of local circuit neurons displayed GABA(B)R1/R2 immunoreactivity. Vesicular glutamate transporters (vGluT1)-containing terminals forming asymmetric synapses expressed GABA(B)R1 and/or displayed postsynaptic GABA(B)R1 at the edges of their asymmetric specialization. Overall, these findings provide evidence for multiple sites where GABA(B) receptors could modulate GABAergic and glutamatergic transmission in the primate CM/PF complex.

Immunohistochemical study of delta and mu opioid receptors on synaptic glomeruli with substance P-positive central terminals in chicken dorsal horn

In an attempt to clarify the mechanism underlying the regulation of the release of substance P (SP) from the central axon terminals of the synaptic glomeruli in lamina II of the dorsal horn, we examined the expression patterns of delta and mu opioid receptors (DOR and MOR) in relation to those of enkephalin (ENK) and SP in the synaptic glomeruli. DOR, MOR, ENK and SP immunoreactivities in lamina II of the dorsal horn in the chicken were examined by confocal laser scanning and electron microscopies. DOR immunoreactivity was localized in both SP-positive central terminals and peripheral elements, while MOR immunoreactivity was only localized in the peripheral elements of the synaptic glomeruli. Both of the peripheral DOR- and MOR-immunoreactive elements were shown to be vesicle-containing dendrites by electron microscopy. Dual immunohistochemistry indicated that DOR, MOR and ENK immunoreactivities were located in distinct peripheral elements. On the basis of present results, the possible roles of DOR and MOR in the regulation of the release of SP from the central axon terminals in the synaptic glomeruli are discussed.

An immunocytochemical study of calbindin-D28K in laminae I and II of the dorsal horn and spinal ganglia in the chicken with special reference to the relation to substance P-containing primary afferent neurons

The localization of calbindin-D28K (CB) was studied immunocytochemically in laminae I and II of the dorsal horn and in spinal ganglia in the chicken, and compared with the distribution of substance P (SP) using double immunolabeling. At the light microscopic level, CB immunoreactivity was observed most intensely in the lamina II using the avidin-biotinylated peroxidase complex (ABC) and immunofluorescence methods. At the electron microscopic level using the ABC method, CB immunoreactivity was observed in the following three neuronal elements: 1) the scalloped central terminal with many dense-cored vesicles (DCVs) in the synaptic glomerulus; 2) some vesicle-containing dendrites (VCDs) inside or outside the synaptic glomerulus; and 3) some axon terminals outside the synaptic glomerulus. The CB-immunoreactive (IR) VCDs in the synaptic glomerulus often formed reciprocal synapses with the central terminal. Strong immunoreactivity was observed at the postsynaptic membrane of CB-IR elements. Double immunofluorescence and immunolabeling methods at the electron microscopic level showed that CB and SP colocalized in the scalloped central terminal with DCVs of the synaptic glomerulus. Almost all SP-IR neurons in the spinal ganglion revealed the coexistence of CB in serial sections in the chicken. In light of previous biochemical and physiological reports, our findings suggest that CB - coexisting with SP - plays an important role in the control of pain transmission through its strong Ca(2+)-buffering action in the chicken.

Pre- and postsynaptic localizations of the CB1 cannabinoid receptor in the dorsal horn of the rat spinal cord

Several lines of evidence show that endogenous and exogenous cannabinoids modulate pain transmission at the spinal level through specific cannabinoid-1 (CB1) receptors. Since anatomical data concerning spinal CB1 receptors are rather contradictory, we studied the cellular and subcellular localizations of the CB1 receptors by immunocytochemistry. Results show a dual pre- and postsynaptic localization of CB1 receptors. Presynaptic receptors are evidenced by the labeling of (1) heterogeneous dorsal root ganglion neurons and (2) axons of Lissauer’s tract. Postsynaptic receptors are shown by the labeling of numerous interneurons in the outer part of lamina II. Double immunolabelings show that lamina II outer CB1 neurons, probably islet cells, may also contain GABA or nitric oxide synthase. Numerous CB1-containing neurons in lamina X are also immunostained with anti-nitric oxide synthase (NOS) antibody. Under the electron microscope, CB1 immunoreactivity is exclusively localized postsynaptically in both somatic and dendritic compartments. The absence of labeling on primary afferent axon terminals is discussed and compared to the absence of labeling on terminals or vesicle-containing dendrites of islet cells, where a presynaptic localization was expected according to data of the literature.

Synaptic architecture of glomeruli in lamina II of the chicken spinal cord, as revealed using ultrathin section and freeze fracture techniques.

Synaptic glomeruli in lamina II of the chicken dorsal horn were studied using the freeze fracture technique, and the results were compared with those obtained using the ultrathin section technique. Our findings using the freeze fracture technique were as follows. (1) On the presynaptic P-face of the central terminal, intramembrane particles (IMPs) were arranged circularly around a small dimple which was reported to be a synaptic vesicle attachment site. A distinct area with aggregated large IMPs was found on the postsynaptic E-face of some peripheral neuronal elements. (2) The area with small IMPs intermingled with several dimples and the area with aggregated large IMPs were present juxtaposed on the same central terminal P-face. The area with aggregated large IMPs indicates that the central terminal functions as a postsynaptic element; accordingly, the two areas represent a reciprocal synapse. (3) Distinct IMP aggregates were observed on the P-face of vesicle-containing dendrites which did not face the central terminal. (4) A fractured septate junction was revealed as numerous parallel-lined furrows on the E-face of the central terminal. The distribution of IMPs in the synaptic glomerulus supports the hypothesis that the synaptic glomerulus is the site of the local inhibitory feedback circuit for pain transmission.

A di-synaptic projection from the superior colliculus to the head of the caudate nucleus via the centromedian-parafascicular complex in the cat: an anterograde and retrograde labeling study

Centromedian-parafascicular (CM-Pf) complex of the thalamus receives inputs from the superior colliculus (SC). The CM-Pf neurons, in turn, project to the neostriatum. In order to establish a circuitry that the efferent terminals of SC neurons make direct synaptic contacts with thalamostriatal neurons in the CM-Pf complex, the anterograde tracer (biocytin) was injected into the unilateral SC and the retrograde tracer (wheat germ agglutinin conjugated to horseradish peroxidase, WGA–HRP) into the ipsilateral head of the caudate nucleus of the cat. The anterogradely labeled SC fibers and their terminals and retrogradely labeled CM-Pf neuronal elements were examined under light and electron microscope. At the light micrsocopic level, biocytin-labeled terminal-like varicosities were observed densely in the latero-dorsal part of the CM and the dorsal part of the Pf lateral to the fasciculus retroflexus. These varicosities were often in close proximity to HRP-labeled somata and dendrites of CM-Pf neurons. Electron microscopic analysis revealed that the biocytin-labeled synaptic boutons contained mainly round synaptic vesicles and established asymmetrical synaptic contacts with retrogradely labeled thalamostriatal neuronal elements, including perikarya, and small and large dendrites. In addition, anterogradely labeled terminals made synaptic contacts with unlabeled somata, small and large dendrites, and spines as well as profiles containing synaptic vesicles. These vesicle-containing profiles were considered to be pre-synaptic elements contacting on thalamostriatal neurons or the vesicle-containing dendrites of local circuit neurons. These results demonstrate that the neurons of the CM-Pf complex of the thalamus is involved in a di-synaptic tecto-striatal circuit.

Ultrastructural localization of P2X3 receptors in rat sensory neurons.

We used isolated IgG antibodies selective for P2X3 receptors to study the ultrastructural distribution of these receptors in rat sensory neurons. In trigeminal ganglia, P2X3 receptor immunoreactivity occurred in small and large nerve cell bodies and their processes. Endoplasmic reticulum and Golgi apparatus were heavily stained; cytoplasmic matrix was faintly to moderately stained. In synaptic glomeruli in lamina II of cervical dorsal horn, P2X3 receptor-immunoreactive core terminals were postsynaptic to unlabelled vesicle-containing dendrites and axons. In the nucleus of the solitary tract, receptor-positive boutons synapsed on dendrites and cell bodies and had complex synaptic relationships with other axon terminals and vesiculated dendrites. These observations identify sites from which ATP could be released to influence sensory signalling within the central nervous system.

GABA-like immunoreactivity in the thalamic nucleus submedius of the cat.

The distribution of GABAergic elements and their synaptic contacts in the nucleus submedius, a specific nociceptive relay in the medial thalamus of the cat, was studied using light and electron-microscopic postembedding immunohistochemical methods. About one-fourth of the neurons in nucleus submedius were GABA immunoreactive. These neurons were generally smaller than the unlabeled neurons and are probably local circuit neurons. Electron microscopy showed GABA immunoreactivity in two types of vesicle-containing profiles, F-terminals and presynaptic dendrites. F-terminals formed simple synapses with the dendrites of presumed thalamocortical relay cells. Presynaptic dendrites were involved in more complex synaptic arrangements that included ascending trigeminothalamic and spinothalamic tract terminals and thalamocortical relay cell dendrites. Analysis of single sections showed that about 40% of the trigeminothalamic and spinothalamic tract terminals, identified by anterograde transport of horseradish peroxidase, were presynaptic to GABAergic presynaptic dendrites. These results show that GABAergic neurons are frequent in nucleus submedius and that the GABAergic elements make synaptic connections similar to those described for other sensory relay nuclei, including the somatosensory ventroposterior nucleus. This suggests that GABAergic mechanisms play an important role in the processing of nociceptive and thermoreceptive information.

GABA and glycine in synaptic glomeruli of the rat spinal dorsal horn.

The superficial dorsal horn of rat spinal cord contains two types of synaptic glomerulus, which are centred around the terminals of unmyelinated and myelinated primary afferents respectively. Both types of glomerulus contain GABAergic axons and dendrites, which are thought to originate from local inhibitory interneurons. Some of the dendrites contain synaptic vesicles, and may be presynaptic to the central axon of dendroaxonic synapses. In order to determine whether GABAergic structures in the two types of glomerulus are derived from different populations of interneurons, a quantitative post-embedding immunogold study of GABA- and glycine-immunoreactivity in a rat dorsal horn was performed. In type I glomeruli, all of the peripheral axons and most vesicle-containing dendrites were GABA-immunoreactive, but only one of 32 axons and none of the vesicle-containing dendrites was glycine-immunoreactive. In contrast, most of the peripheral axons and some of the vesicle-containing dendrites in type II glomeruli possessed both GABA- and glycine-immunoreactivity. This strongly suggests that different types of inhibitory interneuron in the dorsal horn are associated with the two types of glomerulus. It is therefore likely that different populations of interneurons mediate presynaptic inhibition of unmyelinated and myelinated primary afferents.

Post-embedding immunogold labeling of gamma-aminobutyric acid in lamina II of the spinal trigeminal subnucleus pars caudalis: I. A qualitative study.

This study examines the normal synaptic organization of the feline spinal trigeminal nucleus pars caudalis (PC). A primary goal of this study is to identify and characterize the synaptic complexes within PC based on their specific neurotransmitter content. Post-embedding immunogold techniques are utilized with electron microscopy to determine the ultrastructural localization of gamma-aminobutyric acid (GABA) immunoreactivity within lamina II of PC. The colloidal gold particles (10 nm) are randomly distributed over immunoreactive (IR) profiles without preference toward membranous or cytoplasmic regions. GABA immunoreactivity occurs on small unmyelinated axons, on terminals which form synaptic contacts, and on some vesicle-containing dendrites. The GABA-IR terminals form symmetric (type II) contracts onto unlabeled somata and dendrites of various sizes, and onto other unlabeled axon terminals. The GABA-IR terminal in axo-axonic complexes is presynaptic to a round vesicle-containing terminal, which itself may form a type I asymmetric contact onto an unlabeled dendrite or soma. A proportion of vesicle-containing dendrites show GABA-immunoreactivity and are post-synaptic to unlabeled terminals with round vesicles. Other, but far fewer, vesicle-containing dendrites are GABA negative and postsynaptic to GABA-IR terminals. In summary, the findings are consistent with the localization of GABA in intrinsic neurons, and may be associated with presynaptic and postsynaptic inhibition within nociceptive related pathways.

Electron microscopy of immunoreactivity patterns for glutamate and γ‐aminobutyric acid in synaptic glomeruli of the feline spinal trigeminal nucleus (subnucleus caudalis)

We studied the ultrastructure of the synaptic organization in the feline spinal trigeminal nucleus, emphasizing specific neurotransmitter patterns within lamina II of the pars caudalis/medullary dorsal horn. Normal adults were perfused, and Vibratome sections from pars caudalis were processed for electron microscopy. Ultrathin sections were reacted with antibodies for the excitatory neurotransmitter glutamate (Glu) and for the inhibitory neurotransmitter γ-aminobutyric acid (GABA) by using postembedding immunogold techniques. Both single- and double-labeled preparations were examined. Results with single labeling show that Glu-immunoreactive terminals have round synaptic vesicles and form asymmetric synaptic contacts onto dendrites. GABA-immunoreactive axon terminals and vesicle-containing dendrites have pleomorphic vesicles, and the axon terminals form symmetric contacts onto dendrites and other axons. Double labeling on a single section shows glomeruli with central Glu-immunoreactive terminals that are presynaptic to dendrites, including GABA+ vesicle-containing dendrites. These Glu+ terminals are also postsynaptic to GABA+ axon terminals, and these GABA-immunoreactive terminals may also be presynaptic to the GABA+ vesicle-containing dendrites. Quantitative analyses confirm the specificity of the Glu and GABA immunoreactivities seen in the various glomerular profiles. The results suggest that a subpopulation of Glu-immunoreactive primary afferents (excitatory) may be under the direct synaptic influence of a GABA-immunoreactive intrinsic pathway (inhibitory) by both presynaptic and postsynaptic mechanisms. © 1996 Wiley-Liss, Inc.

Electron microscopy of immunoreactivity patterns for glutamate and gamma-aminobutyric acid in synaptic glomeruli of the feline spinal trigeminal nucleus (Subnucleus Caudalis).

We studied the ultrastructure of the synaptic organization in the feline spinal trigeminal nucleus, emphasizing specific neurotransmitter patterns within lamina II of the pars caudalis/medullary dorsal horn. Normal adults were perfused, and Vibratome sections from pars caudalis were processed for electron microscopy. Ultrathin sections were reacted with antibodies for the excitatory neurotransmitter glutamate (Glu) and for the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) by using postembedding immunogold techniques. Both single- and double-labeled preparations were examined. Results with single labeling show that Glu-immunoreactive terminals have round synaptic vesicles and form asymmetric synaptic contacts onto dendrites. GABA-immunoreactive axon terminals and vesicle-containing dendrites have pleomorphic vesicles, and the axon terminals form symmetric contacts onto dendrites and other axons. Double labeling on a single section shows glomeruli with central Glu-immunoreactive terminals that are presynaptic to dendrites, including GABA+ vesicle-containing dendrites. These Glu+ terminals are also postsynaptic to GABA+ axon terminals, and these GABA-immunoreactive terminals may also be presynaptic to the GABA+ vesicle-containing dendrites. Quantitative analyses confirm the specificity of the Glu and GABA immunoreactivities seen in the various glomerular profiles. The results suggest that a subpopulation of Glu-immunoreactive primary afferents (excitatory) may be under the direct synaptic influence of a GABA-immunoreactive intrinsic pathway (inhibitory) by both presynaptic and postsynaptic mechanisms.

Neuronal and synaptic organization of the centromedian nucleus of the monkey thalamus: a quantitative ultrastructural study, with tract tracing and immunohistochemical observations.

The ultrastructure of the centromedian nucleus of the monkey thalamus was analysed qualitatively and quantitatively and projection neurons, local circuit neurons, and synaptic bouton populations identified. Projection neurons were mostly medium-sized, with oval-fusiform or polygonal perikarya, few primary dendrites, and frequent somatic spines; local circuit neurons were smaller. Four basic types of synaptic boutons were distinguished: (1) Small- to medium-sized boutons containing round vesicles (SR) and forming asymmetric contacts, identified as corticothalamic terminals. (2) Heterogeneous medium-sized boutons with asymmetric contacts and round vesicles, similar to the so-called large round (LR) boutons, which were in part of cortical origin. (3) Heterogeneous GAD-positive small- to medium-sized boutons, containing pleomorphic vesicles and forming symmetric contacts (F1 type), which included pallidothalamic terminals. (4) Presynaptic profiles represented by GAD-positive vesicle-containing dendrites of local circuit neurons. Complex synaptic arrangements, serial synapses and triads with LR and SR boutons engaging all parts of projection neuron dendrites and somata, were seen consistently, whereas classical glomeruli were infrequent. LR and SR boutons also established synapses on dendrites of local circuit neurons. F1 boutons established synapses on projection neuron somata, dendrites and initial axon segments. Compared to other previously studied motor-related thalamic nuclei, differences in synaptic coverage between proximal and distal projection neuron dendrites were less pronounced, and the density of synapses formed by local circuit dendrites on projection neuron dendrites was lower. Thus, compared to other thalamic nuclei, the overlap of different inputs was higher on monkey centromedian cells, and centromedian inhibitory circuits displayed a different organization.

Interaction between substance P-immunoreactive central terminals and γ-aminobutyric acid-immunoreactive elements in synaptic glomeruli in the lamina II of the chicken spinal cord

We investigated the interaction between γ-aminobutyric acid (GABA)-immunoreactive (IR) elements and substance P (SP)-IR central terminals in synaptic glomeruli in lamina II of the chicken spinal cord in order to ascertain how pain information is modulated in the spinal dorsal horn. We combined the peroxidase-antiperoxidase (PAP) technique and the protein A-gold (PAG) technique to observe the synaptic relationship between these two components. At the light microscopic level, we observed both GABA-IR and SP-IR elements in the lamina II. GABA-IR elements were also observed in the lamina III. At the electron microscopic level, the following three GABA-IR elements formed synapses with the SP-IR central terminals in synaptic glomeruli: (1) elements which appeared to be axon terminals containing tightly-packed pleomorphic clear vesicles; (2) elements which appeared to be vesicle-containing dendrites with loosely-packed clear and dense-cored vesicles (DCVs); and (3) dendrites without synaptic vesicles. The first type of element was always presynaptic to the SP-IR central terminal. The second type was postsynaptic, presynaptic or in some cases reciprocal to the SP-IR central terminals. The third type was postsynaptic to the SP-IR central terminal. These results suggest that the SP-containing primary afferents activate GABA-containing dendrites and that the SP-containing primary afferents are inhibited presynaptically by GABA-containing neurons through axo-axonic and dendro-axonic synapses.

Cerebellar-responsive neurons in the thalamic ventroanterior-ventrolateral complex of rats: Light and electron microscopy

The morphology and synaptic organization of neurons in the ventroanterior-ventrolateral nucleus of rats was examined using in vivo intracellular staining techniques. Neurons were characterized electrophysiologically based on intrinsic membrane properties and synaptic responses to stimulation of motor cortex and cerebellar nuclei, as described in the companion paper. Cerebellar-responsive neurons were stained intracellularly with either horseradish peroxidase or biocytin. All stained ventroanteriorventrolateral nucleus neurons were identified as thalamocortical neurons on anatomical (and often electrophysiological) grounds, consistent with previous findings that rat ventroanterior-ventrolateral nucleus is interneuron-sparse. Ventroanterior-ventrolateral nucleus neurons had three to eight thick primary dendrites. Proximal dendrites often exhibited a tufted branching pattern, from which many thinner, higher order dendrites arose. Dendrites branched to form a funnel-like infiltration of the neuropil that resulted in a spherical, roughly homogeneous dendritic field. The axon originated from the cell body or a proximal dendrite and coursed laterally and dorsally to innervate motor cortex. One to five axon collaterals were emitted in the rostral dorsolateral sector of the thalamic reticular nucleus; collaterals were not observed in the ventroanterior-ventrolateral nucleus or other nuclei in dorsal thalamus. The synaptic organization of the ventroanterior-ventrolateral nucleus was examined with electron microscopy, including two intracellularly labeled ventroanterior-ventrolateral nucleus neurons that were shown electrophysiologically to receive monosynaptic inputs from the cerebellum. The neuropil of rat ventroanterior-ventrolateral nucleus lacked the complexity and diversity found in corresponding thalamic nuclei of felines and primates, due to the paucity of interneurons. Vesicle-containing dendrites, dendrodendritic synapses and glomeruli were not observed. Three broad classes of presynaptic terminals were identified. (1) Small round boutons: small boutons containing densely-packed, small round vesicles that formed asymmetric synapses predominantly with the distal dendrites of thalamocortical neurons. These were the most prevalent type of bouton in the ventroanterior-ventrolateral nucleus (78% of presynaptic elements) and likely arose from the cerebral cortex. (2) Large round boutons: large terminals with loosely packed small round vesicles that made multiple asymmetric synapses with proximal and intermediate dendrites. Large round boutons comprised 8% of the neuropil, and likely arose from the cerebellar nuclei. (3) Medium size boutons with pleomorphic vesicles: mediumsized profiles containing pleomorphic vesicles that formed symmetric synapses with proximal, intermediate and distal dendrites and, less frequently, with cell bodies. These boutons constituted 14% of the neuropil, and their morphology was similar to previous descriptions of GABAergic immunoreactive profiles in rat ventroanterior-ventrolateral nucleus, as well as boutons originating from the thalamic reticular nucleus (which is the primary source of GABAergic input to the ventroanterior-ventrolateral nucleus). The synaptic organization of thalamocortical neurons and the scarcity of interneurons in the ventroanterior—ventrolateral nucleus is discussed in relation to the critical role of the thalamic reticular nucleus in modulating the efficacy of cortical and cerebellar afferents in the ventroanterior-ventrolateral nucleus and in determining the functional state of ventroanterior-ventrolateral nucleus neurons.

Light- and electron-microscopic analysis of neuropeptide Y-immunoreactive profiles in the cat spinal dorsal horn

The organization of neuropeptide Y-containing profiles in the dorsal horn of cat lumbosacral spinal cord was examined in an immunocytochemical study employing a specific antiserum against neuropeptide Y. Light-microscopic inspection revealed heavy concentrations of immunoreactive axons and varicosities within the superficial layers of the dorsal horn (laminae I and II) and only low to moderate numbers of positive terminals in the deeper layers (laminae III–VI). Neuropeptide-Y immunoreactivity in the superficial laminae occurred primarily as single punctate terminals, although in sagittal sections long rostrocaudally orientated fibres were also found. Immunoreactive fibres in the deeper layers were usually long and beaded. Two-hundred and eight neuropeptide Y-immunoreactive profiles throughout laminae I–VI were examined through serial sections with the electron microscope, and the overwhelming majority ( n = 194) was confirmed to be axon terminals, most of which (95%) formed synaptic junctions. These terminals were packed with small irregularly shaped agranular vesicles, together with a number of large dense-core vesicles. Immunoreactivity was homogenously scattered throughout the cytoplasm, and was also associated with the dense-core vesicles. A few neuropeptide Y-containing profiles ( n = 14) were difficult to classify but they could have been vesicle-containing dendrites. The postsynaptic targets of neuropeptide Y-positive terminals were similar throughout each dorsal horn lamina. Most frequently, neuropeptide Y-positive boutons formed axodendritic and axosomatic synaptic junctions (range = 64% of synapses in laminae V/VI to 83% in lamina III). A smaller proportion of synapses were found upon other axon terminals and in laminae I–III the postsynaptic axon terminals were sometimes the central boutons of glomeruli. A number of terminals, especially those in lamina II, formed multiple synapses which often comprised a triadic arrangement. These findings suggest that neuropeptide Y regulates spinal sensory transmission through both a postsynaptic action upon dorsal horn neurons and a presynaptic action upon primary afferent terminals.

Light and electron microscopic localization of calcitonin gene‐related peptide immunoreactivity in lamina II of the feline trigeminal pars caudalis/medullary dorsal horn: A qualitative study

Calcitonin gene-related peptide (CGRP) is a neuropeptide that is associated with a subset of primary afferent fibers and appears to have a role in nociception. The purpose of the present study was to perform a qualitative light, and especially electron microscopic (LM and EM), examination of CGRP-immunoreactivity (IR) within lamina II (substantia gelatinosa) of the feline pars caudalis/medullary dorsal horn (PC/MDH) of the spinal trigeminal nucleus. The LM investigation revealed massive CGRP-IR within lamina II outer, with fewer fibers that traversed lamina II inner. The EM preparations showed CGRP-IR in small, thinly myelinated and unmyelinated axons, preterminal axons, and in axon terminals that formed asymmetric synaptic contacts onto small dendritic profiles. The terminals with CGRP-IR were often the central element within glomeruli, where the terminal usually formed 2 or more asymmetric synaptic specializations onto 1 or more dendrites. Many of these postsynaptic dendrites contained synaptic vesicles. Other profiles were seen forming presynaptic contacts onto the terminal with CGRP-IR, and these profiles most likely represent presynaptic dendrites and/or other axon terminals of intrinsic origin. The synaptic association of terminals showing CGRP-IR with vesicle-containing dendrites, presynaptic dendrites, and/or other axon terminals suggests that terminals with CGRP-IR are especially susceptible to modulation.

Ultrastructural and immunocytochemical study of lamina II islet cells in rat spinal dorsal horn

In order to compare the ultrastructure of GABA-immunoreactive and nonimmunoreactive islet cells in lamina II of the rat dorsal horn, a combined ultrastructural and immunocytochemical study of nine Golgi-stained neurones was performed. Cell bodies of these neurones were tested with antiserum to GABA, and in most cases with antiserum to glycine, while parts of the cell body and dendritic tree were examined with the electron microscope. Four of the neurones had cell bodies that were immunoreactive with GABA antiserum, and 2 of these were also glycine-immunoreactive, while 2 were not. Cell bodies of the remaining five neurones were not immunoreactive with GABA antiserum, nor, in the 3 cases tested, with glycine antiserum. Three of the GABA-immunoreactive cells possessed vesicle-containing dendrites and were presynaptic at dendrodendritic synapses, whereas no vesicles were observed in the dendrites of any of the neurones that were not GABA-immunoreactive. The axon of one of the nonimmunoreactive cells was found with the electron microscope. It gave rise to boutons that contained round agranular vesicles and a few dense-cored vesicles. Three synapses formed by this axon were identified and all were asymmetric. No obvious differences were detected in the types of profile that were presynaptic to GABA-immunoreactive and nonimmunoreactive cells. These results suggest that GABAergic islet cells are a source of presynaptic dendrites in lamina II of the rat and that some presynaptic dendrites contain GABA and glycine, while others contain GABA without glycine. The nonimmunoreactive islet cells presumably represent a distinct functional class of neurones and some of these may release an excitatory amino acid transmitter, possibly in addition to one or more neuropeptides.