STT Cells Research Articles

Neurological mechanisms of chest pain and cardiac disease.

SCS is an efficacious, reversible, and safe therapy that improves quality of life, increases exercise tolerance, and relieves angina pectoris, but clinical trials in North America are needed to confirm the data coming from Europe. Neuronal convergence onto STT cells underlies the referred pain associated with angina pectoris. With pain referred to the chest and upper arm, cardiac nociceptive information is transmitted via sympathetic afferent fibers to thoracic cells. With pain referred to the jaw and neck, cardiac nociceptive information is transmitted via vagal afferent fibers onto cervical cells. SCS can modulate the responses of thoracic STT cells to nociceptive input originating from the heart. SCS modulates cardiac function. It stabilizes neurons in the intrinsic cardiac nervous system, and can reduce infarct size via adrenoreceptors.

Pain pathways: Comparison of monkey laminae I and V STT cells with human pain sensation

The aim of this study was to compare the responses of monkey laminae I and V STT cells to repeated brief-contact heat pain and the thermal grill illusion of pain with human psychophysics. Single L5-S1 STT units are antidromically identified and characterized in barbiturate-anesthetized macaques. Repeated brief-contact (0.7 s) heat stimuli are applied at 48, 51, 54C and 5, 3, 2 s ISIs. Reset is tested by omitting 1-2 stimuli in a longer series. The thermal grill is applied with a series of parallel 2mm bars at 40/20C. All HPC lamina I STT cells tested showed augmentation, reset, and grill responses corresponding to human sensation. In contrast, NS lamina I and WDR lamina V STT cells showed little or no augmentation, reset, or grill responses. These observations support the concept that HPC lamina I STT cells have a unique role as a virtual labeled line for second pain (Craig, Ann Rev Neurosci 26:1-30, 2003). Dysfunction of HPC cells could be important for neuropathic pain.

Responses of Spinothalamic Lamina I Neurons to Repeated Brief Contact Heat Stimulation in the Cat

It was recently shown that repeated heat stimulation, using brief contacts (<1 s) with a preheated thermode at sufficiently short interstimulus intervals (ISIs <5 s) and high temperatures (> or =51 degrees C), will elicit in humans a sensation of rapidly augmenting "second" (burning) pain with only a weak "first" (sharp) pain sensation. Most strikingly, at short intertrial intervals (ITIs >5 s) such summation will reset, or begin again at baseline. In the present experiments, the responses of nociceptive lamina I spinothalamic (STT) neurons in the lumbosacral dorsal horn of barbiturate-anesthetized cats were examined using this repeated brief contact heat paradigm. The neurons were classified as nociceptive-specific (NS, n = 8) or polymodal nociceptive (HPC, n = 8) based on their responses to quantitative thermal stimuli; all had receptive fields on the glabrous ventral hindpaw. A pneumatic piston was used to apply a thermode preheated to 34, 46, 49, 53, or 58 degrees C with a contact dwell time of approximately 0.7 s to the ventral hindpaw repeatedly (15 times) at ISIs of 2, 3, and 5 s, with 3-5 min between trials. The mean responses of the 16 nociceptive lamina I STT cells showed rapid temporal summation that was directly dependent on temperature and inversely dependent on ISI, with the greatest increases occurring between the 3rd and 10th contacts. The temporal profiles of this family of curves correspond with the psychophysical data on human sensation. Further analysis showed that this summation was due to the HPC cells, which all showed strong summation; in contrast, the NS cells showed little, if any. The HPC responses to the repeated heat stimuli lagged each contact by approximately 1 s, consistent with the strong, monosynaptic C-fiber input that is characteristic of HPC cells and also with the dependence of second pain on C-fiber nociceptors. HPC cells also displayed the reset phenomenon at short ITIs, again in correspondence with the psychophysical data. The summation and the reset displayed by HPC cells were not related to skin temperature. Thus the results presented in this study, together with those in the preceding article, demonstrate a double dissociation indicating that NS and HPC lamina I STT cells can subserve the qualitatively distinct sensations of first (sharp) and second (burning) pain, respectively. These findings support the concept that the lamina I STT projection comprises several discrete sensory channels that are integrated in the forebrain to generate distinct sensations.

Association of spinothalamic lamina I neurons and their ascending axons with calbindin-immunoreactivity in monkey and human

The calbindin-immunoreactivity of spinothalamic (STT) lamina I neurons and their ascending axons was examined in two experiments. In the first experiment, lamina I STT neurons in macaque monkeys were double-labeled for calbindin and for retrogradely transported WGA∗HRP following large ( n=2) or small ( n=1) injections that included the posterior thalamus. Most, but not all (78%) of the contralateral retrogradely labeled lamina I STT cells were positive for calbindin. Calbindin-immunoreactivity was not selectively associated with any particular anatomical type of lamina I STT cell; 82% of the fusiform cells, 78% of the pyramidal cells and 67% of the multipolar cells were double-labeled. In the second experiment, oblique transverse sections from upper cervical spinal segments of three macaque monkeys, one squirrel monkey and five humans were stained for calbindin-immunoreactivity. In each case, a distinct bundle of fibers was densely stained in the middle of the lateral funiculus. This matches the location of anterogradely labeled ascending lamina I axons observed in prior work in cats and monkeys, and it matches the location of the classically described ‘lateral spinothalamic tract’ in humans. This bundle had variable shape across cases, an observation that might have clinical significance. These findings support the view that lamina I STT neurons are involved in spinal cordotomies that reduce pain, temperature and itch sensations.

Quantitative response characteristics of thermoreceptive and nociceptive lamina I spinothalamic neurons in the cat.

The physiological characteristics of antidromically identified lamina I spinothalamic (STT) neurons in the lumbosacral spinal cord were examined using quantitative thermal and mechanical stimuli in barbiturate-anesthetized cats. Cells belonging to the three main recognized classes were included based on categorization with natural cutaneous stimulation of the hindpaw: nociceptive-specific (NS), polymodal nociceptive (HPC), or thermoreceptive-specific (COOL) cells. The mean central conduction latencies of these classes differed significantly; NS = 130.8 +/- 55.5 (SD) ms (n = 100), HPC = 72.1 +/- 28.0 ms (n = 128), and COOL = 58.6 +/- 25.3 ms (n = 136), which correspond to conduction velocities of 2.5, 4.6, and 5.6 m/s. Based on recordings made prior to any noxious stimulation, the mean spontaneous discharge rates of these classes also differed: NS = 0.5 +/- 0.7 imp/s (n = 47), HPC = 0.9 +/- 0.7 imp/s (n = 59), and COOL = 3.3 +/- 2.6 imp/s (n = 107). Standard, quantitative, thermal stimulus sequences applied with a Peltier thermode were used to characterize the stimulus-response functions of 76 COOL cells, 47 HPC cells, and 37 NS cells. The COOL cells showed a very linear output from 34 degrees C down to approximately 15 degrees C and a maintained plateau thereafter. The HPC cells showed a fairly linear but accelerating response to cold below a median threshold of approximately 24 degrees C and down to 9 degrees C (measured at the skin-thermode interface with a thermode temperature of 2 degrees C). The HPC cells and the NS cells both showed rapidly increasing, sigmoidal response functions to noxious heat with a fairly linear response between 45 and 53 degrees C, but they had significantly different thresholds; half of the HPC cells were activated at ~45.5 degrees C and half of the NS cells at approximately 43 degrees C. The 20 HPC lamina I STT cells and 10 NS cells tested with quantitative pinch stimuli showed fairly linear responses above a threshold of approximately 130 g/mm(2) for HPC cells and a threshold of approximately 100 g/mm(2) for NS cells. All of these response functions compare well (across species) with the available data on the characteristics of thermoreceptive and nociceptive primary afferent fibers and the appropriate psychophysics in humans. Together these results support the concept that these classes of lamina I STT cells provide discrete sensory channels for the sensations of temperature and pain.

Cooling-specific spinothalamic neurons in the monkey.

1. Little is known concerning the processing of innocuous thermoreceptive information in the CNS of the monkey. The aim of the present study was to confirm the prediction, based on recent studies in cat and monkey, that there must be a prominent spinothalamic (STT) projection of cooling-specific spinal cord lamina I neurons to the posterior part of the ventral medial nucleus (VMpo) of the monkey thalamus. 2. Experiments were performed on four cynomolgus monkeys anesthetized with pentobarbital sodium. A detailed mapping of somatosensory thalamus was performed in each animal, and VMpo was identified by recordings from clusters of thermoreceptive-specific and nociceptive-specific (NS) neurons. Stimulating electrodes were then implanted in VMpo. Tungsten microelectrodes were used to record the responses of neurons in the superficial dorsal horn of the lumbosacral spinal cord. 3. Many spontaneously active lamina I neurons were found that were inhibited by radiant warming and that responded to innocuous cooling of the hindpaw. These cooling-specific (COLD) neurons were excited by small temperature drops below skin temperature and increased their discharge with decreasing skin temperature. They were not excited by thermally neutral mechanical stimuli applied to the receptive fields. In passing, we also characterized with natural stimulation a few NS neurons reponsive to pinch and/ or noxious heat, multimodal (HPC) neurons responsive to noxious heat, pinch, and cold stimuli, and wide-dynamic-range neurons responsive to both innocuous and noxius cutaneous stimuli that were encountered in lamina I. 4. Twenty lamina I COLD cells were identified as STT neurons by antidromic activation from the contralateral VMpo. The mean conduction latency for these units was 26.1 ms, which corresponds to a mean conduction velocity of approximately 8.0 m/s. They were not antidromically activated from an electrode in the region of the ventral posterior nucleus in the thalamus. In addition, we antidromically activated from VMpo four NS units and three HPC cells. 5. These findings demonstrate for the first time the existence of a prominent direct projection of specific COLD lamina I STT cells to thalamus in the monkey. This is consistent with clinical inferences in humans and with prior results in cats. This result confirms that the dense lamina I STT projection to VMpo demonstrated in anatomic studies includes COLD cells, and it supports the role of VMpo as a thalamic relay nucleus for pain- and temperature-related information.

Glutamatergic and GABAergic input to rat spinothalamic tract cells in the superficial dorsal horn

The distribution of synaptic terminals onto spinothalamic tract cells (types I and II) of the superficial dorsal horn was determined with special reference to the amino acid transmitters glutamate and gamma-aminobutyric acid. Fifteen spinothalamic cells retrogradely labeled from the thalamus with the neuroanatomical tracer wheatgerm agglutinin conjugated to horseradish peroxidase were sectioned for electron microscopy. Serial sections from several levels through each cell were immunostained for glutamate and gamma-aminobutyric acid using a postembedding immunogold technique. Perimeter measurements of spinothalamic cell somata and dendrites and the lengths of apposition for all terminal profiles in contact with the spinothalamic cells were obtained from electron micrographs using a digitizing tablet. These data were used to determine the density of terminals on the soma and dendrites. In addition, the terminal population on these cells was categorized by transmitter content (glutamate, gamma-aminobutyric acid, or unlabeled). The results demonstrate that terminal density increased on dendrites relative to their distance from the soma. Glutamatergic and GABAergic input composed 37% and 20% of the terminal population, respectively, and these percentages remained uniform for the soma and dendrites. There were no significant differences among the 15 cells analyzed for this study. The results, therefore, suggest that both type I and type II STT cells of the superficial DH have similar synaptic organizations.

The effect of phorbol esters on the responses of primate spinothalamic neurons to mechanical and thermal stimuli

1. Sensitization of dorsal horn neurons is thought to play an important role in pain perception, secondary hyperalgesia, and allodynia. Recent experimental evidence suggests that the sensitization of dorsal horn neurons is induced by combined increased release of excitatory amino acids and peptides in the spinal cord dorsal horn from nociceptive primary afferents due to an injury-caused barrage of impulses. We tested the hypothesis that protein kinase C (PKC) is involved as a second messenger in this process of neuronal sensitization. To activate PKC, infusion of a phorbol ester [12-O-tetradecanoylphorbol-13-acetate (TPA)] into the dorsal horn through a microdialysis fiber was used. During TPA infusion the background activity of spinothalamic (STT) neurons increased substantially. After TPA application, while the background activity of the STT neurons was still increased, the responses evoked by either innocuous or noxious mechanical stimulation of the cutaneous receptive field did not change from the control level. However, 1 h after TPA administration the background activity returned to the control level and responses to innocuous mechanical stimuli were significantly elevated. The responses of STT cells to noxious heat and noxious mechanical stimuli did not change significantly after TPA administration. When a phorbol ester that does not activate PKC was applied (alpha-TPA), no significant changes in background or evoked activity of STT cells were observed. Our results provide evidence that PKC may play an important role in the process of sensitization of dorsal horn neurons to innocuous mechanical stimuli.

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.

Structure-function relationships in identified afferent neurones

The review deals with structure-function relationships in primary afferent and spinal cord neurones that were intracellularly injected with a marker substance (mostly HRP) after physiological identification. At the level of dorsal root ganglion (DRG) cells, there is a significant correlation between soma size and conduction velocity (or diameter) of the afferent fibre for most subpopulations of DRG cells, but the scatter of data is considerable, so that the size of a DRG cell soma cannot be predicted from the diameter of its axon or vice versa. The spinal terminations of primary afferent fibres are the best example of a relationship between structure and function, since most of the afferent units possess characteristic patterns of spinal arborization, e.g. the "flame-shaped arbors" of hair follicle afferents in lamina III of the dorsal horn, or the projection of nociceptive afferents onto lamina I. The morphological features of spinal cord neurones can be used only to a limited extent for functional identification. Thus, many SCT neurones can be recognized by their triangular dendritic tree and STT cells in lamina VII/VIII by their dendritic projection into the white matter. It is still not possible, however, to distinguish a nociceptive STT cell from a low-threshold mechanoreceptive one on the basis of morphological criteria.

Primate spinothalamic pathways: I. A quantitative study of the cells of origin of the spinothalamic pathway

In six monkeys spinothalamic (STT) cells were retrogradely labeled by injecting 2% wheat germ agglutinin-conjugated horseradish peroxidase into the somatosensory thalamus. Following a 5-day survival period, the animals were perfused and the tissue was removed and processed with the tetramethyl benzidine technique. In all animals there were HRP-labeled STT cells in all segments of the spinal cord. In one old world monkey, the injection included most of the thalamus and resulted in 18.235 estimated total number of STT cells. Of this total, 35% were located in the upper cervical segments (C1-C3), 18% were located in C4-C8, 19% were in the thoracic spinal cord with most found in T1-T3; 6% were in L1-L3, 13% were in L4-L7, and 7% were in the coccygeal segments. Of the total labeled STT cells, 17% were found in the spinal cord ipsilateral to the thalamic injections; 53% of these cells were located in C1-C3 primarily in lamina VIII. The percentage of label found in the contralateral lower cervical region laminae I-III (43-50%), IV-VI (33-48%), and VII-X (8-17%) was similar among three animals with similar thalamic injections. The distributions of the shapes of the labeled STT cells were similar for each lamina between the lower cervical and lower lumbar regions. The mean diameter of the labeled STT cells varied with spinal cord segment and lamina. The lamina I STT cells were the smallest. In the cervical spinal cord, lamina VIII STT cells had the largest diameters, while in the lumbar region laminae IV-VI had the largest STT cells.

Origin of thalamically projecting somatosensory relay neurons in the immature rat

The locations of spinothalamic (STT) and trigeminothalamic (TTT) neurons in 14–18-day rats using retrograde transport of fluorescent latex microspheres were examined. The aim of this study was to determine whether the connections between the somatosensory relay neurons and the thalamus were established in these immature rats. The majority of the labeled STT and TTT neurons was found in the brainstem and upper cervical cord (C 1–C 4). These cells were distributed to a number of distinct groups. Among them, the nucleus of trigeminal spinal tract interpolaris (SP5I) had the highest number of TTT cells, and the internal basilar nucleus (IB) comprised the largest population of STT cells. Except for the cells located in the dorsal portion of the ventral horn (VHd), most of the labeled STT and TTT cells were contralateral to the thalamic injection sites. This pattern of distribution of the projecting neurons in immature rats is very similar to the pattern observed in adult rats. The development of the STT and TTT projections in 14–18 day rats is therefore largely complete. This result will allow us to extrapolate our understanding of the membrane properties of projection cells obtained in the immature rats to the adult rats.

Central organization of sympathetic cardiovascular response to pain.

Figure 1 summarizes the pathways discussed in this review. Noxious and innocuous cardiac and somatic information converge on SRT neurons in the upper thoracic spinal cord; the third thoracic segment (T3) is shown in Figure 1. The SRT pathway and collaterals of the STT convey this information to the MRF (mainly RGC). This input is integrated with other inputs (eg. auditory and visual) by MRF neurons, some of which project to the spinal cord. RS motor neurons (dotted pathway) might mediate motor responses to cardiac pain, as well as motor responses associated with escape or alerting behavior. RS neurons (cross-hatched and clear pathways) can also modulate ascending traffic by altering activity of SRT and STT cells. Finally, RS neurons (dotted pathway) might modify sympathetic function via collaterals to IML or to interneurons, or MRF cells could project to medullary areas possessing neurons (solid pathway) that project directly to IML. The small figurines represent the heart (top), blood vessels (middle right) and muscle (bottom).

Effects of intracardiac bradykinin on T2-T5 medial spinothalamic cells.

Effects of injecting bradykinin (2 micrograms/kg) into the left atrium on spinothalamic tract neurons projecting to medial thalamus (M-STT cells), to the ventral posterior lateral nucleus of the thalamus (L-STT cells), or to both (LM-STT cells) were examined in 18 monkeys (Macaca fascicularis) anesthetized with alpha-chloralose. Bradykinin increased cell activity in 11/16 M-STT cells, 10/15 L-STT cells, and 4/7 LM-STT cells. One M-STT cell was inhibited. Peak responses to bradykinin of the three cell groups were not different. LM-STT cells began to respond and reached peak responses slightly earlier than the other two groups. Six M-STT, four L-STT, and two LM-STT cells became entrained to the cardiac cycle during their responses to bradykinin. Responses to bradykinin were not dependent on the type of somatic input or cell location. Responding cells most often received A delta- and C-fiber sympathetic input, but some responding cells had only A delta-input. These results demonstrate that in addition to L-STT cells STT cells projecting to the medial thalamus respond to a potentially noxious cardiac stimulus. These cells may participate in the motivational-affective component of cardiac pain.

The effects of chronic section of the lateral spinothalamic tract on the responses of ventral spinothalamic tract neurons in the cat

Kerr's hypothesis (J. Comp. Neurol., 159 (1975) 335–356) that chronic lesion of the lateral spinothalamic tract (LSTT) might cause the ventral STT (VSTT) to take over the nociceptive function of the LSTT was tested electrophysiologically in 4 cats. Recordings were made from 15 antidromically identified deep STT cells in L7 6 months after lesion of the LSTT at L1, confirming that these cells do not project rostrally in the LSTT. The receptive fields of 12 deep STT cells were studied and found to be characteristic of these cells in normal animals; they had not developed the nociceptive characteristics of lamina LSTT cells. Immunohistochemistry showed that substance P-containing afferents were still contained within the dorsal laminae of the L7 grey matter. These results disprove Kerr's hypothesis and suggest that plasticity in other ascending systems must be responsible for the return of pain after lesion of the LSTT.

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.