Neurons in Rexed's layer II were physiologically characterized with natural and electrical stimuli applied to their cutaneous receptive fields. The neurons were then intracellularly stained with horseradish peroxidase. Three general patterns of physiological responses were found. Nociceptive specific neurons did not respond to gentle mechanical stimulation. Most responded exclusively to tissue-damaging stimuli. Some also responded to moderately heavy pressure, but these responded to noxious stimuli with an increased discharge frequency. Wide dynamic range neurons responded to both gentle mechanical stimulation and to tissue-damaging stimulation. Low-threshold mechanoreceptive neurons responded only to gentle mechanical stimulation. Some of the low-threshold mechanoreceptive neurons were innervated by primary afferents with unmyelinated axons. Excepting those low-threshold mechanoreceptive neurons with input from unmyelinated afferents, the patterns of primary afferents innervation of layer II neurons were similar to the patterns of innervation that have been found for neurons in layers I and IV-V. All but 2 of the 22 neurons that we found were recognized as being of two general morphological types. Stalked cells had their perikarya situated along the superficial border of layer II. Most of their dendrites traveled ventrally while spreading out rostrocaudally. This gave their dendritic arbors a fan-like shape. Stalked cell axons arborized largely in layer I. Islet cell perikarya were found throughout layer II. Most of their dendrites traveled rostrocaudally. Their dendritic arbors were shaped like cylinders with their long axes parallel to the long axis of the spinal cord. Islet cell axons arborized in the immediate vicinity of their dendritic territories, within layer II. Stalked cells and those islet cells whose dendritic arbors were largely contained within the superficial one-third of layer II (layer IIa) were either nociceptive specific or wide dynamic range neurons. The islet cells whose dendritic arbors were largely within the deeper two-thirds of layer II (layer IIb) were all low-threshold mechanoreceptive neurons. These observations suggest that layers IIa and IIb have different functional roles and that stalked cells and islet cells are separate and distinct components of the neural circuitry of the superficial dorsal horn.

In view of continuing uncertainties concerning the organization, afferent inputs, and projection sites of neurons in the subnucleus interpolaris of the trigeminal (V) spinal tract nucleus, the characteristics of 222 single neurons in and adjacent to the subnucleus were examined electrophysiologically in adult cats anesthetized with chloralose. Neurons were tested for orthodromic responsiveness to a variety of stimuli that included nonnoxious tactile stimuli, noxious mechanical and radiant-heat stimuli, and graded electrical stimulation of the skin, mucosa, tooth pulp, and masseter nerve. Antidromic activation techniques were also used to determine if the functionally identified neurons projected directly to the contralateral posterior thalamus, ipsilateral cerebellum, or cervical spinal cord. In addition, the periaqueductal gray matter (PAG) was stimulated to test for conditioning influences from the PAG on orthodromic responses to noxious and nonnoxious oral-facial stimuli. Interpolaris neurons were somatotopically arranged in subnucleus interpolaris in a pattern conforming in general to the medially facing, inverted-head representation characteristic of other parts of the V brain stem sensory nuclear complex. On the basis of their responsiveness to cutaneous stimuli, the neurons could be functionally classified as either cutaneous nociceptive or low-threshold mechanoreceptive (LTM) neurons. The LTM neurons constituted the major neuron type, accounting for over 75% of our neuron sample. Most of them had a localized mechanoreceptive field of less than 100 mm2 in area that was restricted to one V division, and they had skin-evoked response latencies indicative of afferent input predominantly from A-beta cutaneous afferents. A population of nociceptive neurons was also encountered in the lateral, marginal region of interpolaris and at its medial or ventral border with the reticular formation. These neurons were of two types: nociceptive-specific (NS) neurons, which did not respond to nonnoxious stimuli but which required noxious stimuli for their activation; and wide dynamic range ( WDR ) neurons, which responded to both noxious and nonnoxious stimuli applied to the facial skin. Most had an ipsilateral receptive field that was greater than 100 mm2 in area and that often involved two or three V divisions. Their properties generally conformed to those previously described for nociceptive neurons in the medullary dorsal horn (V subnucleus caudalis) and spinal cord dorsal horn. Interpolaris neurons of all classes (LTM, WDR , and NS) were found to have direct axonal projections to the thalamus, cerebellum, and spinal cord.(ABSTRACT TRUNCATED AT 400 WORDS)

Two major interneurons of the outer part of Rexed's layer I1 (IIa) were impaled with microelectrodes, had their primary inputs characterized, and were subsequently filled with horseradish peroxidase. Their fine structural characteristics and synaptic connections were then analyzed electron microscopically. Two islet cells, whose rostrocaudally oriented dendrites were largely confined within layer IIa, received primary input from small myelinated axons. A stalked cell, whose cell body was situated on 1/11 border had a cone-shaped dendritic arbor which traversed layer IIa as well as the inner part of Rexed's layer I1 (IIb) and rostrocaudal dendritic branches which ran for part of their course along the UIIa border. It received primary input from small myelinated as well as from unmyelinated axons. Both cell types received asymmetrical axodendritic synapses from primary endings in layer IIa and IIb glomeruli and widely separated symmetrical axodendritic synapses from small nonprimary endings outside of glomerli.The presence of aggregates of synaptic vesicles in the dendrites of the layer IIa islet cells but not in the stalked cell dendrites constitutes the major fine structural difference between these interneurons. Islet cell dendrites form symmetrical synapses on several different kinds of neural processes. They usually send either a single type 2 spine (spines which contain synaptic vesicles) or dendritic shaft into layer IIa and IIb glomeruli, where they form dendrodendritic synapses on adjacent type 1 spines (spines without synaptic vesicles) and on other small dendritic shafts. Some islet cell type 2 spines also form dendroaxonic synapses on the primary endings.Outside of the glomeruli, islet cell dendrites also form dendrodendritic synapses on type 1 spines and different sized dendritic shafts. They often approach other dendritic shafts forming small bundles of dendrites in which they are reciprocally linked by dendrodendritic synapses to other synaptic vesicle-containing dendrites. At bead-like enlargements of their dendritic shafts and along some of their fine caliber dendritic shafts, the islet cells form dendroaxonic synapses on the shafts of unmyelinated axons. The unmyelinated axon of the islet cell forms symmetrical synapses on layer I1 dendritic shafts and spines outside of glomeruli. The role of the layer IIa islet cell as an inhibitory interneuron is discussed.

During orthodontic tooth movement, the activation of the vascular system in the compressed periodontal ligament (PDL) is an indispensable process in tissue remodeling. We hypothesized that compressive force would induce angiogenesis of PDL through the production of vascular endothelial growth factor (VEGF). We examined the localization of VEGF in rat periodontal tissues during experimental tooth movement in vivo, and the effects of continuous compressive force on VEGF production and angiogenic activity in human PDL cells in vitro. PDL cells adjacent to hyalinized tissue and alveolar bone on the compressive side showed marked VEGF immunoreactivity. VEGF mRNA expression and production in PDL cells increased, and conditioned medium stimulated tube formation. These results indicate that continuous compressive force enhances VEGF production and angiogenic activity in PDL cells, which may contribute to periodontal remodeling, including angiogenesis, during orthodontic tooth movement.