Ultrastructural Morphology, Synaptic Relationships, and CGRP Immunoreactivity of Physiologically Identified C-Fiber Terminals in the Monkey Spinal Cord

The spinal cord terminations of two electrophysiologically identified single C-fibers (one identified as a C-nociceptor) were intra-axonally labeled with horseradish peroxidase and analyzed with both light and electron microscopy. Serial section ultrastructural analysis and postembedding immunocytochemical techniques for calcitonin gene-related peptide (CGRP), substance P (SP), and GABA were used to study the synaptology, and neuropeptide content. All C-terminal synapses were in laminae I and II. The terminals sampled (n = 73) from these two C-fibers rarely established glomerular synaptic complexes, but rather, simple terminals, usually measuring 1–4 μm in length and 1–3 μm in diameter. They most often established 1 or 2 (range 1 to 5) quite large asymmetric axodendritic synaptic contacts. Postsynaptic structures included dendritic spines and shafts with and without vesicles. C-terminals were filled with small round synaptic vesicles (45–60 nm) and also contained variable numbers of large dense-core vesicles (LDCVs, 80–110 nm). LDCVs inside identified C-terminals frequently displayed CGRP immunoreactivity. We were unable to detect SP immunoreactivity inside our sample of C-fiber LDCVs. C-terminals were never found postsynaptic to other profiles. Thus, the C-fiber terminals sampled in this study have simple synaptology, do not receive presynaptic control and contain CGRP immunoreactivity. They differ greatly from the terminals of Aδ nociceptors studied previously by our group that had glomerular endings, often received presynaptic input and did not contain CGRP immunoreactivity. This suggests the existence of different processing mechanisms, at the level of the first synapse, for nociceptive inputs arriving to lamina I and II through different types of primary afferents. © 1993 Wiley-Liss, Inc

Synaptic Interactions Between GABA-Immunoreactive Profiles and the Terminals of Functionally Defined Myelinated Nociceptors in the Monkey and Cat Spinal Cord

This study analyzes the synaptic interactions between the central terminals of A delta high threshold mechanoreceptors (A delta HTMs) and GABA-immunoreactive profiles. A delta HTM primary afferents from three monkeys and one cat were electrophysiologically identified and intracellularly labeled with HRP, and their terminal arborizations in laminae I and II of the sacrocaudal spinal cord were studied at the ultrastructural level. GABA-immunoreactive profiles in relation to A delta HTM terminals were demonstrated using postembedding colloidal gold techniques. Monkey A delta HTM terminals (n = 131) usually constituted the central element of synaptic glomeruli; they established large asymmetric synaptic contacts with 1-13 dendrites (modal value 2- 4) and were surrounded by 0-6 peripheral axon terminals (modal value 2- 3). The large majority (around 85%) of the peripheral axon terminals were GABA immunoreactive. They were found presynaptic to the A delta HTM terminal and/or to dendrites postsynaptic to the primary afferent terminal. Furthermore, all peripheral axon terminals found presynaptic to the A delta HTM terminals showed GABA immunoreactivity. Within a single A delta HTM fiber, this synaptic arrangement was found in 20-60% of its boutons. In addition, 28% of the postsynaptic dendritic profiles displayed weak GABA immunoreactivity. Some of them contained vesicles; however, only in a few cases did we observe synapses between a GABA- immunoreactive vesicle-containing dendrite and a dendritic profile postsynaptic to an A delta HTM terminal. Similar synaptology and interactions with GABA-immunoreactive profiles were displayed by the terminals of the single cat A delta HTM fiber studied. Our data support the hypothesis that GABA-containing neurons use both presynaptic and/or postsynaptic mechanisms to exert a powerful control, presumably inhibitory, over the transmission of nociceptive information between A delta HTM afferents and second-order neurons in monkey and cat spinal cord. Our results also imply that GABA may be released within the synaptic glomeruli formed by A delta HTM terminals either by local dendrites or by axon terminals. We discuss the possibility that these GABAergic synapses can be driven by inputs from both primary afferents and/or descending systems to modulate the transmission of nociceptive sensory information

Ultrastructural Morphology, Synaptic Relationships, and CGRP Immunoreactivity of Physiologically Identified C-Fiber Terminals in the Monkey Spinal Cord

The spinal cord terminations of two electrophysiologically identified single C-fibers (one identified as a C-nociceptor) were intra-axonally labeled with horseradish peroxidase and analyzed with both light and electron microscopy. Serial section ultrastructural analysis and postembedding immunocytochemical techniques for calcitonin gene-related peptide (CGRP), substance P (SP), and GABA were used to study the synaptology, and neuropeptide content. All C-terminal synapses were in laminae I and II. The terminals sampled (n = 73) from these two C-fibers rarely established glomerular synaptic complexes, but rather, simple terminals, usually measuring 1–4 μm in length and 1–3 μm in diameter. They most often established 1 or 2 (range 1 to 5) quite large asymmetric axodendritic synaptic contacts. Postsynaptic structures included dendritic spines and shafts with and without vesicles. C-terminals were filled with small round synaptic vesicles (45–60 nm) and also contained variable numbers of large dense-core vesicles (LDCVs, 80–110 nm). LDCVs inside identified C-terminals frequently displayed CGRP immunoreactivity. We were unable to detect SP immunoreactivity inside our sample of C-fiber LDCVs. C-terminals were never found postsynaptic to other profiles. Thus, the C-fiber terminals sampled in this study have simple synaptology, do not receive presynaptic control and contain CGRP immunoreactivity. They differ greatly from the terminals of Aδ nociceptors studied previously by our group that had glomerular endings, often received presynaptic input and did not contain CGRP immunoreactivity. This suggests the existence of different processing mechanisms, at the level of the first synapse, for nociceptive inputs arriving to lamina I and II through different types of primary afferents. © 1993 Wiley-Liss, Inc

Synaptic Interactions Between GABA-Immunoreactive Profiles and the Terminals of Functionally Defined Myelinated Nociceptors in the Monkey and Cat Spinal Cord

This study analyzes the synaptic interactions between the central terminals of A delta high threshold mechanoreceptors (A delta HTMs) and GABA-immunoreactive profiles. A delta HTM primary afferents from three monkeys and one cat were electrophysiologically identified and intracellularly labeled with HRP, and their terminal arborizations in laminae I and II of the sacrocaudal spinal cord were studied at the ultrastructural level. GABA-immunoreactive profiles in relation to A delta HTM terminals were demonstrated using postembedding colloidal gold techniques. Monkey A delta HTM terminals (n = 131) usually constituted the central element of synaptic glomeruli; they established large asymmetric synaptic contacts with 1-13 dendrites (modal value 2- 4) and were surrounded by 0-6 peripheral axon terminals (modal value 2- 3). The large majority (around 85%) of the peripheral axon terminals were GABA immunoreactive. They were found presynaptic to the A delta HTM terminal and/or to dendrites postsynaptic to the primary afferent terminal. Furthermore, all peripheral axon terminals found presynaptic to the A delta HTM terminals showed GABA immunoreactivity. Within a single A delta HTM fiber, this synaptic arrangement was found in 20-60% of its boutons. In addition, 28% of the postsynaptic dendritic profiles displayed weak GABA immunoreactivity. Some of them contained vesicles; however, only in a few cases did we observe synapses between a GABA- immunoreactive vesicle-containing dendrite and a dendritic profile postsynaptic to an A delta HTM terminal. Similar synaptology and interactions with GABA-immunoreactive profiles were displayed by the terminals of the single cat A delta HTM fiber studied. Our data support the hypothesis that GABA-containing neurons use both presynaptic and/or postsynaptic mechanisms to exert a powerful control, presumably inhibitory, over the transmission of nociceptive information between A delta HTM afferents and second-order neurons in monkey and cat spinal cord. Our results also imply that GABA may be released within the synaptic glomeruli formed by A delta HTM terminals either by local dendrites or by axon terminals. We discuss the possibility that these GABAergic synapses can be driven by inputs from both primary afferents and/or descending systems to modulate the transmission of nociceptive sensory information