Haploinsufficiency of protamine-1 or -2 causes infertility in mice.

Protamines are the major DNA-binding proteins in the nucleus of sperm in most vertebrates and package the DNA in a volume less than 5% of a somatic cell nucleus. Many mammals have one protamine, but a few species, including humans and mice, have two. Here we use gene targeting to determine if the second protamine provides redundancy to an essential process, or if both protamines are necessary. We disrupted the coding sequence of one allele of either Prm1 or Prm2 in embryonic stem (ES) cells derived from 129-strain mice, and injected them into blastocysts from C57BL/6-strain mice. Male chimeras produced 129-genotype sperm with disrupted Prm1 or Prm2 alleles, but failed to sire offspring carrying the 129 genome. We also found that a decrease in the amount of either protamine disrupts nuclear formation, processing of protamine-2 and normal sperm function. Our studies show that both protamines are essential and that haploinsufficiency caused by a mutation in one allele of Prm1 or Prm2 prevents genetic transmission of both mutant and wild-type alleles.

Spinothalamic tract neurons in the substantia gelatinosa.

The substantia gelatinosa of the mammalian spinal cord is generally believed to be a closed system; that is its neurons are thought to project only to the substantia gelatinosa of the same or the contralateral side. Experiments in monkeys, using injections of the marker enzyme horseradish peroxidase, show that at least some neurons of the substantia gelatinosa project to the thalamus and thus belong to the spinothalamic tract. Such neurons include two cell types intrinsic to the gelatinosa, the central cells and the limitrophe cells of Cajal.

Activation of protein kinase B/Akt in the periphery contributes to pain behavior induced by capsaicin in rats.

Protein kinase B (PKB/Akt) is a member of the second-messenger regulated subfamily of protein kinases. It is implicated in signaling downstream of growth factors, insulin receptor tyrosine kinases and phosphoinositide 3-kinase (PI3K). Current studies indicate that nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and PI3K help mediate inflammatory hyperalgesia. However, little is known about the role of PKB/Akt in the nociceptive system. In this study, we investigated whether PKB/Akt in primary sensory neurons is activated after noxious stimulation and contributes to pain behavior induced in rats by capsaicin. We demonstrated that phospho-PKB/Akt (p-PKB/Akt) is increased in dorsal root ganglia (DRG) at 5 min after intradermal injection of capsaicin. p-PKB/Akt is distributed predominantly in small- and medium-sized DRG cells. After capsaicin injection, p-PKB/Akt (473) is colocalized with isotectin-B4 (IB4), tyrosine kinase A (TrkA), and calcitonin gene-related peptide (CGRP). Furthermore, most transient receptor potential vanilloid type 1 (TRPV1) positive DRG neurons double label for p-PKB/Akt. Behavioral experiments show that intradermal injection of a PI3K (upstream of PKB/Akt) inhibitor, wortmannin, dose-dependently inhibits the changes in exploratory behavior evoked by capsaicin injection. The PKB/Akt inhibitor, Akt inhibitor IV, has the same effect. The results suggest that the PKB/Akt signaling pathway in the periphery is activated by noxious stimulation and contributes to pain behavior.

Antagonism of morphine analgesia by nonopioid cold-water swim analgesia: direct evidence for collateral inhibition.

The demonstrated existence of opioid and nonopioid forms of pain control has raised questions as to how they interact. Previous indirect evidence suggests that activation of one system inhibited the activation of the other. The present study assessed this directly using morphine as an opiate form of analgesia and continuous cold-water swims (CCWS, 4 degrees C, 2 min) as the nonopioid form. A significant reduction in morphine (8 mg/kg, SC) analgesia on the tail-flick test was observed if rats were acutely exposed to CCWS immediately prior to morphine administration. The inability of naloxone (10 mg/kg, SC) to reduce CCWS analgesia verified its nonopioid nature. The antagonism of morphine (3 mg/kg, SC) analgesia was greater following preexposure to 2 min of CCWS than 1 min of CCWS. CCWS was also more effective in antagonizing analgesia induced by the 3 mg/kg than the 8 mg/kg dose of morphine. The antagonism of morphine analgesia by CCWS was dependent upon the temporal patterning of stimulus presentation: exposure to CCWS 20 or 60 min prior to morphine failed to alter subsequent morphine analgesia. A significant reduction in analgesia induced by intraperitoneal administration of morphine (10 mg/kg) was also observed when CCWS was presented immediately prior to injection, suggesting that pharmacokinetic factors such as altered drug absorbance by CCWS-induced vasoconstriction do not appear to explain these effects. These data provide direct support for the existence of collateral inhibitory mechanisms activated by CCWS and morphine, and suggests that these opioid and nonopioid forms of analgesia do not function synergistically, but instead involve some form of hierarchical order.

Collaterals of spinothalamic cells in the rat.

Spinothalamic (STT) cells were investigated in the rat to determine the distribution of subpopulations with terminals in both the lateral and medial thalamus, the thalamus bilaterally, or the thalamus and the medullary reticular formation. Two or more retrogradely transported substances (fluorescent dyes, and/or horseradish peroxidase) were injected in each animal. Three combinations of injections were most commonly used: (1) injections of the medullary reticular formation and thalamus, (2) separate injections into each side of the thalamus, and (3) separate injections into the medial and lateral thalamus. The distribution of single labeled cells after each injection was compared with previously published results for rats. The distribution of cells which contained both tracers, double-labeled (DL) cells, was the focus of this study. An average of 15% of STT cells and 8% of spinoreticular cells projected to both the reticular formation and thalamus. However, only a small component of STT cells (less than 2%) projected bilaterally into the thalamus. Most DL cells were found in upper cervical segments. The laminar distribution of all three groups of DL neurons were similar. These cells were most often located in the reticulated part of lamina V and the intermediate zone, lamina VII. STT cells that had terminals in both the medial and lateral thalamus and STT cells with collaterals in the reticular formation were concentrated on the side contralateral to their terminals. These DL neurons provide an anatomical substrate for noxious stimuli to stimuli to activate the reticular formation and thalamus and/or specific sensory and intralaminar thalamus simultaneously.

Distribution of the postsynaptic dorsal column projection in the cuneate nucleus of monkeys.

Cells in the spinal cord that are postsynaptic to primary afferent fibers project to the dorsal column nuclei in the postsynaptic dorsal column pathway. The projection of cells in the cervical spinal cord of monkeys to the cuneate nucleus has been reported to avoid pars rotunda of that nucleus, the part that contains the somatotopic representation of the ipsilateral hand. We used the sensitive anterograde tracer Phaseolus vulgaris leucoagglutinin to reexamine this projection. We made multiple iontophoretic injections into the cervical enlargements of three monkeys (two Macaca fascicularis and one Macaca mulatta). Control injections were made in the contralateral dorsal columns of one of these and in the dorsal roots of a fourth animal (M. fascicularis) to test for transport by fibers of passage. After 28-39 days, the animals were deeply anesthetized and perfused, and the tissue was processed for immunohistochemical detection of the label. In all cases (excluding control injections), labeled fibers and varicosities were distributed widely in the ipsilateral cuneate and external cuneate nuclei, including pars rotunda. The dorsal column nuclei ipsilateral to control injections contained no label or only very few poorly labeled fibers, indicating that labeling through fibers of passage did not contribute importantly to the results. This study indicates that the postsynaptic projection to the cuneate nucleus is widespread and includes pars rotunda. Such projections may contribute to transmission of information originating in nociceptors through the dorsal column-medial lemniscal system to the ventrobasal thalamus.

The organization of the electromotor nucleus and extraocular motor nuclei in the stargazer (Astroscopus y-graecum).

The organization of the oculomotor and electromotor systems was examined in the stargazer, a teleost. The electromotor system in these animals is a derivative of the oculomotor system. The extraocular motor nuclei and nerves consist of approximately equal numbers of motoneurons and axons (about 100 per muscle). In contrast, electromotor axons appear to branch several times within the intracranial portion of the IIIrd nerve. The topographical organization of the motoneurons was examined using retrograde transport of horseradish peroxidase injected into the electric organ or eye muscles. Electromotor and oculomotor neurons form distinct populations. Each electric organ receives a strong ipsilateral and a weak contralateral innervation. Individual eye muscles receive unilateral innervations with the expected laterality. Within the oculomotor nucleus there is some topographical separation of motoneurons innervating each muscle. Antidromic field potentials confirm the identity of the electromotor nucleus.

Glutamate-immunoreactive terminals synapse on primate spinothalamic tract cells.

Glutamate has been shown to excite spinothalamic tract (STT) neurons and has been localized to primary afferent neurons, spinal cord projection neurons, and interneurons in the spinal cord dorsal horn. The likelihood that glutamate-immunoreactive (GLU-IR) terminals directly innervate STT neurons was investigated. For these studies three lamina IV or V STT cells in the lumbar spinal cords of three monkeys (Macaca fascicularis) were identified electrophysiologically and characterized. Two were identified as high threshold neurons and one as a wide dynamic range neuron. Following intracellular injection of the cells with HRP and reaction to give the cells a Golgi-like appearance, the tissues were processed for electron microscopy. Postembedding immunogold methods with antibodies specific for glutamate were used to identify GLU-IR terminals apposing the somata and dendrites of the STT neurons, including dendrites that extended into laminae IV and III. The GLU-IR terminals were numerous and constituted a mean of 46% of the population counted that appose the STT soma and 50% of the profiles apposing the dendrites. Fifty-four percent of the somatic and 50% of the dendritic surface length was contacted by GLU-IR terminals. Most terminals contained round clear vesicles and some contained a variable number of large dense core vesicles. For one of the three cells examined it was determined that 45% of the terminals apposing the soma were GLU-IR and 30% of the terminals were gamma aminobutyric acid-immunoreactive (GABA-IR). In an additional monkey, a lamina I cell retrogradely labeled from the ventral posterolateral nucleus of the thalamus was found to be ensheathed in glial processes.(ABSTRACT TRUNCATED AT 250 WORDS)

GABA-immunoreactive terminals synapse on primate spinothalamic tract cells.

Gamma-aminobutyric acid (GABA) is a putative inhibitory neurotransmitter in the vertebrate nervous system. Several lines of evidence suggest that GABA plays an important role in the processing and modulation of sensory input in the spinal cord dorsal horn. In the present study, the relationship between GABA-immunoreactive (GABA-IR) terminals and spinothalamic tract (STT) cells in the monkey lumbar cord was investigated. Physiologically characterized STT cells, one located in lamina V and two located in lateral lamina IV, were intracellularly injected with horseradish peroxidase (HRP). A fourth STT cell, located in lamina I, was retrogradely labeled following injection of HRP into the contralateral thalamus. Immunogold labeling of ultrathin sections through the cell bodies and proximal dendrites of the STT neurons demonstrated that the percentage of the GABA-IR terminals in contact with these profiles was 24.7% and 36%, respectively. The average STT surface length contacted by GABA-IR terminals for cell bodies and proximal dendrites was 18.2% and 26.7%, respectively. For the lamina I cell, 7 out of 35 (20%) of the terminals were GABA-IR and they covered 9.6% of the surface analyzed. These data demonstrate that GABA-IR terminals synapse directly on STT cells, constituting a substantial proportion of the terminal population on these cells. Furthermore, compared to the cell bodies, a greater percentage of the input on the proximal dendrites is GABAergic. These anatomical data are consistent with the findings of a previously published iontophoretic study that demonstrated that GABA can exert a strong inhibitory influence on STT cells. These findings are discussed in relation to GABAergic involvement in tonic and phasic inhibition of STT neurons.

The cells of origin of the primate spinothalamic tract.

Spinothalamic tract cells in the lumbar, sacral and caudal segments of the primate spinal cord were labelled by the retrograde transport of horseradish peroxidase (HRP) injected into the thalamus. The laminar distribution of stained spinothalamic cells in the lumbosacral enlargement differed according to whether the HRP was injected into the lateral or the medial thalamus. Lateral injections labelled cells in most laminae, but the largest numbers of cells were in laminae I and V. The highest concentrations of cells labelled from the medial thalamus were in laminae VI-VIII. Ninety percent or more of the stained spinothalamic cells in the lumbosacral enlargement were contralateral to the injection site. In the conus medullaris stained spinothalamic cells were most numerous in laminae I, V and VI following lateral thalamic injections of HRP. Many of the cells of the conus were in Stilling's nucleus. Twenty-three percent of the cells in the conus were ipsilateral to the injection site in the lateral thalamus. Only a few cells in the conus were labelled by medial thalamic injections. The total number of spinothalamic cells from L5 caudally was estimated to be at least 1,200-2,500. An injection of HRP into the midbrain resulted in laminar distribution of labelled cells much like that produced by a lateral thalamic injection. The types of spinothalamic tract cells and the sizes of their somata were determined for different laminae. The cell types resemble those already described from Golgi and other studies of the spinal cord gray matter. The spinothalamic tract cells in lamina I included Waldeyer cells and numerous small fusiform, pyriform or triangular cells. Those in lamina II included limitrophe and central cells. Spinothalamic cells in lamina III were central cells. Most of the labelled cells in laminae IV-X were polygonal, although there were also flattened cells in these layers. The smallest spinothalamic cells were in laminae I-III, while the largest were in laminae V and VII-IX. Spinothalamic cells in the conus medullaris included cells like those in the lumbosacral enlargement, but also a special cell type in Stilling's nucleus. Some cells in the conus had dendrites that crossed the midline. Spinothalamic axons could sometimes be traced to the ventral white commissure within one or a few sections. In longitudinal sections, most labelled axons were in the ventral part of the lateral funiculus on the side of the injection, although a few were in the ventral funiculus or on the contralateral side. The axons were widely dispersed, and a few were located adjacent to the pia-glial membrane.

Differential projections of cat medullary raphe neurons demonstrated by retrograde labelling following spinal cord lesions.

Neurons of the medullary raphe nuclei in cats were retrogradely labelled following injection of horseradish peroxidase (HRP) into the L6 spinal cord segment. Brainstems were cut in sagittal section to facilitate examination of the rostral-caudal extent of raphe neurons projecting to the spinal cord. Large numbers of HRP-labelled neurons were found in nucleus raphe magnus, nucleus raphe pallidus, and nucleus raphe obscurus (as well as a few neurons in nucleus raphe pontis). Dorsal or ventral hemisections at the T12-L1 level restricted HRP retrograde transport to those pathways within the intact portion of spinal cord, allowing a determination of the part of the cord through which raphe neurons project to the lumbar enlargement. Neurons of nucleus raphe magnus were found to project primarily in dorsolateral fasciculus. A significant number of neurons of nucleus reticularis gigantocellularis also project in dorsolateral fasciculus. Nucleus raphe obscurus neurons were found to project primarily in ventral funiculus, while nucleus raphe pallidus neurons project in the ventrolateral fasciculi and ventral funiculus. The serotonergic (5HT) fibers described by Dahlström and Fuxe ('65) to terminate in the dorsal horn, intermediolateral cell column, and ventral horn are likely to coincide with the raphe-spinal projections documented in this work.

Direct catecholaminergic innervation of primate spinothalamic tract neurons.

Catecholaminergic axonal varicosities identified by immunocytochemical staining for dopamine-beta-hydroxylase were observed at the light microscopic level apposing the somata of retrogradely labeled spinothalamic tract neurons in the monkey spinal cord. Three retrogradely labeled and two intracellularly labeled spinothalamic neurons were serially sectioned and examined at selected intervals at the electron microscopic level. Electron microscopic study revealed that axonal boutons directly contacted the somata and/or dendrites of lamina I, IV, and V spinothalamic tract neurons. All of the profiles apposing one of the retrogradely labeled lamina I spinothalamic tract neurons were categorized from eight planes of section spaced at 1-micron intervals. Of the 305 profiles counted that were adjacent to this soma, 17 (5.6%) stained positively for dopamine-beta-hydroxylase. Of these 17 appositions, three were followed in serial sections to confirm that they had synaptic thickenings and alignment of vesicles along the membrane contacting the spinothalamic tract soma. Catecholaminergic boutons were observed apposing the somata and dendrites of intracellularly filled STT cells characterized as high threshold and wide dynamic range neurons. These observations clearly indicate a direct innervation of spinothalamic tract neurons by catecholaminergic neurons, providing anatomical data to support previous physiological findings demonstrating that catecholamines modulate nociceptive transmission.

Electrophysiological response properties of spinoreticular neurons in the monkey.

Extracellular recordings were made from 29 spinoreticular cells in the spinal cords of anesthetized monkeys. The cells were in either the cervical or the lumbar enlargement, and they were identified by antidromic activation from the medial part of the pontomedullary reticular formation. More spinoreticular neurons were sampled in the cervical than in the lumbar cord. Most of the cells were contralateral to the side from which antidromic activation was observed, but a higher proportion of the spinoreticular neurons in the cervical enlargement than in the lumbar enlargement was ipsilateral to the antidromic stimulus. Three cells in the lumbar cord were antidromically activated not only from the reticular formation but also from the contralateral thalamus, confirming that some spinoreticular projections are formed by collaterals from spinothalamic cells. Most of the spinoreticular neurons were in the ventral horn in laminae VII and VIII, although a few were in laminae IV-VI. Nearly half of the spinoreticular cells in the sample could not be activated by any form of peripheral stimulation tested. The other cells could be activated by stimulation of receptive fields that varied from small to large, that were sometimes bilateral regions of the skin or deep tissues. Although some spinoreticular cells could be classified as low threshold or wide dynamic range, the largest proportion were high threshold, requiring noxious stimulation for their activation. Descending volleys resulting from stimulation in the reticular formation could often be shown to inhibit or to excite spinoreticular neurons. It can be concluded that at least some spinoreticular neurons may play a role in nociception.

Organization of spinothalamic tract axons within the rat spinal cord.

Two techniques have been used to examine the organization of spinothalamic tract axons within the spinal cord of the rat. In the initial experiments, the thalamus was filled on one side with horseradish peroxidase (HRP) using a series of small injections. The injections were preceded by lesions of various areas of the ventral quadrant. These studies indicated that the cells of origin of STT axons ascending within the ventral funiculus (VF) are located primarily in the ventral-most areas of the dorsal horn and the intermediate gray zone. The cells of origin of STT axons projecting within the ventrolateral funiculus (VLF) are located not only deep within the gray matter but in addition within the dorsal-most two thirds of the dorsal horn, the area of the spinal cord gray matter shown in previous studies to contain the vast majority of cells with cutaneous tactile and nociceptive input. To examine these projections directly, rats received either a series of HRP injections that filled the thalamus on one side or a small injection into either medial or lateral thalamus. Examination of the labeled axons in horizontal sections through the cervical cord indicated that STT axons ascending to lateral thalamus do so in the VLF. In contrast, axons terminating in medial thalamus ascend in the VF. Additional experiments have shown that axons ascending to the lateral thalamus are distributed throughout the VLF at lumbar levels. Within the thoracic cord, lateral projecting SST axons are distributed throughout much of the VLF but are not found in close proximity to the ventral horn. At cervical levels all lateral-projecting STT axons have assumed a position on the lateral rim of the VLF. These and previously published data have demonstrated that the rat spinothalamic tract is composed of two components that differ in the distribution of their cells of origin, the area of the cord in which they ascend, and the thalamic nuclei in which they terminate.

Mapping study of serotoninergic input to diencephalic-projecting dorsal column neurons in the rat.

Several lines of evidence indicate that the processing of somatosensory information in the dorsal column nuclei (DCN) is subject to descending controls. Anatomical experiments have demonstrated projections to the DCN from the sensorimotor cerebral cortex and the reticular formation. Physiological studies have shown that the activity of DCN neurons can be altered following stimulation of the cerebral cortex, reticular formation, periaqueductal gray, or raphe nuclei. Recent biochemical and electrophysiological evidence suggests a serotoninergic modulation of DCN neurons. The present study identifies serotonin-containing contacts on cells in the DCN that project to the thalamus in the rat. Retrograde labeling of brainstem neurons by horseradish peroxidase demonstrated projections to the DCN from the nucleus reticularis paragigantocellularis lateralis and from several raphe nuclei, including nuclei raphe obscurus (RO), pallidus (RP), and magnus (RM). Double labeling with horseradish peroxidase and antibody for serotonin indicated that the RO, RP and RM are likely to be the sources of the serotoninergic projections to the DCN. Thus, the role of the serotoninergic output from the raphe nuclei includes modulation of activity in the DCN.

Ascending projections from the area around the spinal cord central canal: A Phaseolus vulgaris leucoagglutinin study in rats.

A single small iontophoretic injection of Phaseolus vulgaris leucoagglutinin labels projections from the area surrounding the spinal cord central canal at midthoracic (T6-T9) or lumbosacral (L6-S1) segments of the spinal cord. The projections from the midthoracic or lumbosacral level of the medial spinal cord are found: 1) ascending ipsilaterally in the dorsal column near the dorsal intermediate septum or the midline of the gracile fasciculus, respectively; 2) terminating primarily in the dorsal, lateral rim of the gracile nucleus and the medial rim of the cuneate nucleus or the dorsomedial rim of the gracile nucleus, respectively; and 3) ascending bilaterally with slight contralateral predominance in the ventrolateral quadrant of the spinal cord and terminating in the ventral and medial medullary reticular formation. Other less dense projections are to the pons, midbrain, thalamus, hypothalamus, and other forebrain structures. Projections arising from the lumbosacral level are also found in Barrington's nucleus. The results of the present study support previous retrograde tract tracing and physiological studies from our group demonstrating that the neurons in the area adjacent to the central canal of the midthoracic or lumbosacral level of the spinal cord send long ascending projections to the dorsal column nucleus that are important in the transmission of second-order afferent information for visceral nociception. Thus, the axonal projections through both the dorsal and the ventrolateral white matter from the CC region terminate in many regions of the brain providing spinal input for sensory integration, autonomic regulation, motor and emotional responses, and limbic activation.

Funicular trajectories of brainstem neurons projecting to the lumbar spinal cord in the monkey (Macaca fascicularis): a retrograde labeling study.

Brainstem nuclei projecting to the lumbar spinal cord in the monkey were identified by using horseradish peroxidase and the fluorescent dye granular blue. These retrogradely transported tracers were used in fluid and/or gel forms to determine the funicular trajectories of the brainstem-spinal projections. The major descending components of the dorsal funiculus arose from the n. gracilis, n. cuneatus, and the n. of the solitary tract. Major components of the dorsolateral funiculus (DLF) came from the raphe complex, medullary and pontine reticular formation, locus coeruleus, Edinger-Westphal n., and red n. Other nuclei giving rise to minor contributions to the DLF included n. gracilis, n. cuneatus, n. of the solitary tract, medial and spinal vestibular n., subcoeruleus, periaqueductal gray, interstitial n. of Cajal, n. of Darkschewitsch, and the anteromedian n. The major components of ventral cord paths (ventrolateral and ventral funiculi) arose from the raphe complex, the medullary and pontine reticular formation, lateral and spinal vestibular n., and the coerulean complex. Minor contributions to the ventral paths descended from the dorsal motor n. of X, n. of the solitary tract, medial vestibular n., paralemniscal reticular formation, dorsal parabrachial n., n. cuneiformis, periaqueductal gray, Kölliker-Fuse n., and red n. The possible functional implications of the funicular distribution of these descending pathways are discussed from the perspective of descending inhibition and pain modulation.

Unmyelinated preganglionic fibers in frog ventral roots.

It has been suggested that the preganglionic visceral fibers in the frog are unmyelinated. To obtain further evidence for this suggestion: (1) the percentages of unmyelinated fibers and the myelinated fiber histograms were done for all frog ventral roots, (2) the number of unmyelinated fibers were counted on either side of a chronic section of the seventh ventral root and (3) compound action potentials were obtained for roots 7-10. The results are that slightly more than 80% of the unmyelinated fibers in the seventh ventral root arise in the spinal cord and are presumably unmyelinated preganglionic efferents. Furthermore, a C fiber volley could be followed from the seventh ventral root into caudal parts of the sympathetic chain. Thus this data provides confirmation for the idea that many frog preganglionic fibers are unmyelinated. This study also shows that 24% of the ventral root axons in the frog are unmyelinated and that, on the basis of percentages of unmyelinated fibers and myelinated fiber histograms, there are five categories of frog ventral roots.

Collaterals of primate spinothalamic tract neurons to the periaqueductal gray.

Collateral projections are an important feature of the organization of ascending projections from the spinal cord to the brain. Primate spinothalamic tract (STT) neurons with collaterals to the periaqueductal gray (PAG) were studied by means of a fluorescent double-labeling method. Granular Blue and rhodamine-labeled latex microspheres were placed in the ventral posterior lateral (VPL) nucleus of the thalamus and the periaqueductal gray, respectively. Single and double labeled neurons were studied in the upper cervical cord, cervical enlargement, thoracic cord, lumbar enlargement, and sacral segments. The laminar distribution of double labeled neurons was similar to that of spinomesencephalic tract (SMT) neurons. Most double labeled (STT-SMT) neurons were located in contralateral laminae I, V, VII, and X. Relatively more lamina I STT-SMT neurons were found in the cervical enlargement and more lamina V STT-SMT neurons in the lumbar enlargement. The density of STT-SMT neurons in the upper cervical segments and cervical enlargement was almost equal. The density of STT-SMT neurons in the lumbar enlargement was 40% of that in the cervical enlargement. The thoracic and sacral segments had the lowest density of STT-SMT neurons, about 10% of that in the cervical enlargement. STT-SMT neurons constituted 14.7% of SMT neurons and 6% of STT neurons in the cervical enlargement and 15.3% of SMT neurons and 2.9% of STT neurons in the lumbar enlargement. The branch points of eight STT-SMT axons were studied electrophysiologically. The average percentage of conduction time spent in the parent axon was more than 85% for an antidromic action potential from the VPL nucleus and 91% from the PAG. Branch points of STT-SMT axons were calculated to be 9-13 mm caudal to the PAG, in the pons or rostral medulla.

Nuclei in which functionally identified spinothalamic tract neurons terminate.

The approximate level of termination of the axons of individual, functionally characterized spinothalamic tract neurons within the monkey thalmus was mapped by antidromic activation using a monopolar electrode which was moved in a systematic grid of tracks through the thalamus. The course of individual axons could be followed through several thalamic levels, and in a few cases branches to both the VPL nucleus and to the intralaminar nuclei were demonstrated. Most of the axons studied, however, projected just to the VPLc or VPLo nuclei. The spinothalamic tract cells that projected to the VPLc nucleus included representative of all known functional categories: low threshold, wide dynamic range, high threshold and "deep." It is speculated that these different classes of spinothalamic projections could make contributions to such sensory modalities as touch, proprioception and pain.