Periaqueductal Gray Stimulation Research Articles

Lombard reflex during PAG-induced vocalization in decerebrate cats

The Lombard reflex occurs when a speaker increases his vocal effort while speaking in the presence of ambient noise. The purpose of this study was to clarify whether the Lombard reflex can be evoked during controlled vocalization in an animal model. In decerebrate cats, repetitive electrical stimulation was applied to the midbrain periaqueductal gray (PAG) to evoke vocalization. Pure tone auditory stimulation was delivered through a loudspeaker. The activities of the laryngeal adductor muscle, diaphragm and external oblique abdominal muscle and the voice intensity were measured during PAG stimulation, in the presence and absence of the auditory stimulation. To clarify the effects of the auditory–laryngeal reflex on the activity of laryngeal adductor motoneurons, the amplitude of the laryngeal reflex evoked by single shock stimulation of the superior laryngeal nerve was also measured during respiration, in the presence and absence of auditory stimulation. The sound made by the cats due to PAG-induced vocalization was augmented by exposure to auditory stimulation, and the activities of the laryngeal adductor muscle and external oblique abdominal muscle were also augmented. During respiration, auditory stimulation also increased the amplitude of the laryngeal reflex evoked in the laryngeal adductor muscle. These results demonstrate that the essential neuronal mechanisms for evoking the Lombard reflex exist within the brainstem.

Chronic spinal nerve ligation induces changes in response characteristics of nociceptive spinal dorsal horn neurons and in their descending regulation originating in the periaqueductal gray in the rat.

We studied whether a chronic neuropathy induced by unilateral spinal nerve ligation changes the response characteristics of spinal dorsal horn wide-dynamic range (WDR) neurons or their periaqueductal gray (PAG)-induced descending modulation. Experiments were performed in rats with behaviorally demonstrated allodynia induced by spinal nerve ligation and in a group of nonneuropathic control rats. The stimulus-response functions of WDR neurons for mechanical and thermal stimuli and the modulation of their peripherally evoked responses by electrical stimulation of the PAG were determined under pentobarbital anesthesia. The results showed that neuropathy caused a significant leftward shift in stimulus-response functions for mechanical stimuli. In contrast, stimulus-response functions for noxious heat stimuli in the neuropathic limb were, if anything, shifted rightward, although this shift was short of statistical significance. In neuropathic rats, PAG stimulation produced a significantly stronger attenuation of spinal neuronal responses induced by noxious heat in the unoperated than in the operated side. At the intensity that produced attenuation of noxious heat stimuli, PAG stimulation did not produce any significant change in spinal neuronal responses evoked by mechanical stimuli either from the operated or the nonoperated hindlimb of the neuropathic rats. Spontaneous activity of WDR neurons was higher in the operated side of neuropathic rats than in control rats. Afterdischarges evoked by peripheral stimuli were observed in 1/16 of the WDR neurons ipsilateral to spinal nerve ligation and not at all in other experimental groups. The WDR neurons studied were not activated by innocuous or noxious cold stimuli. The results indicate that spinal nerve ligation induces increased spontaneous activity and enhanced responses to mechanical stimuli in the spinal dorsal horn WDR neurons, whereas noxious heat-evoked responses are not significantly changed or if anything, attenuated. Moreover, the inhibition of noxious heat stimuli by PAG stimulation is attenuated in the neuropathic side. It is proposed that the observed changes in the response characteristics of the spinal dorsal horn WDR neurons and in their descending modulation may contribute to the neuropathic symptoms in these animals.

Role of pulmonary afferent inputs in vocal on-switch in the cat.

The purpose of this paper is to elucidate the possible role of pulmonary afferent inputs in triggering inspiratory-vocal (I-V) and Vocal-inspiratory (V-I) transitions during periaqueductal gray (PAG)-induced vocalization. Under ketamine anesthesia, we investigated the effects of changes in pulmonary afferent inputs on the PAG-induced vocal motor pattern by transection and electrical stimulation of the cervical vagus nerve in non-paralyzed cats and by lung inflation and deflation in paralyzed cats. After bilateral vagotomy, PAG stimulation induced apneusis; strong suppression of the I-V transition disrupted the vocal rhythmicity. Electrical stimulation of the central end of the cut vagus nerve during PAG stimulation immediately caused an I-V transition. In paralyzed cats during the withholding of lung inflation, the I-V transition was also suppressed during PAG stimulation. Lung inflation during PAG stimulation caused a phase switch from inspiration to fictive vocalization, i.e. I-V transition. In contrast, this fictive vocal phase maintained by lung inflation was terminated by lung deflation, i.e. V-I transition. These findings suggest that pulmonary vagal afferent feedback plays an important role in triggering and terminating vocalization.

The nucleus retroambigualis controls laryngeal muscle activity during vocalization in the cat.

The purpose of this study was to determine (1) whether the nucleus retroambigualis (NRA) plays an essential role in periaqueductal gray (PAG)-induced vocalization and (2) which NRA neurons are involved in the projection from the PAG to laryngeal motoneurons. Bilateral injections of the neurotoxin kainic acid into the NRA in decerebrate cats abolished PAG-induced vocalization; PAG stimulation after the injections no longer modulated vocal fold adductor or tensor activity, and only tonically, but no longer phasically, activated the abdominal muscles. In contrast, PAG-induced inspiratory excitation remained even after the injections. These results suggest that the NRA is essential for the vocal activation of the laryngeal adductor and abdominal muscles, and that an additional pathway from the PAG to respiratory motoneurons other than through the NRA is important for mediating PAG-induced inspiratory activation. Secondly, axonal projections of NRA neurons to the contralateral nucleus ambiguus (NA) were studied electrophysiologically. Five expiratory neurons, which had decrementing (n=4) or constant (n=1) firing patterns, were identified as both having axonal projections to the NA and receiving inputs from the PAG. Furthermore, following NA stimulation many constant-latency action potentials of silent cells were recorded in the vicinity of the NRA, where many silent cells were also orthodromically activated by PAG stimulation. No NRA augmenting expiratory neurons could be antidromically activated from the NA. It is suggested that the NRA and adjacent reticular formation integrate inputs from the PAG and send outputs to laryngeal motoneurons for vocalization.

SEROTONERGIC Pontomedullary Neurons Are Not Activated by Antinociceptive Stimulation in the Periaqueductal Gray

The antinociceptive and cardiovascular effects of midbrain periaqueductal gray (PAG) stimulation are mediated through a relay in the pontomedullary raphe magnus (RM) and adjacent nucleus reticularis magnocellularis (NRMC). To test whether the neurons important in mediating PAG-evoked effects are SEROTONERGIC, the responses of pontomedullary SEROTONERGIC-LIKE cells to PAG stimulation were tested. SEROTONERGIC-LIKE neurons (n = 21) were recorded extracellularly in halothane-anesthetized Sprague Dawley rats. Serotonergic-like neurons were distinguished by their slow and steady background discharge. Two neurons that were physiologically characterized as SEROTONERGIC-LIKE were intracellularly labeled and processed for serotonin immunoreactivity; both cells tested contained immunoreactive serotonin. Train stimulation of sites within the midbrain PAG, at intensities of </=50 microA, suppressed the tail withdrawal from noxious heat and evoked changes in blood pressure and heart rate. No SEROTONERGIC-LIKE cells were activated by single-pulse or short-train (two to five pulses) stimulation of the PAG at antinociceptive intensities. In most cases, SEROTONERGIC-LIKE cells were unaffected by long-train stimulation (5-6 sec) of the PAG, which produced antinociception and cardiovascular changes. In contrast, >50% of the cells in two nonserotonergic-like cell classes were activated at short latency by such PAG stimulation. In conclusion, monosynaptic excitation of SEROTONERGIC cells in RM/NRMC is unlikely to be necessary for the nociceptive and autonomic modulatory effects of PAG stimulation.

Inhibitory effects evoked from both the lateral and ventrolateral periaqueductal grey are selective for the nociceptive responses of rat dorsal horn neurones

In rats anaesthetized with alphaxalone/alphadolone a comparative study was made of the inhibitory effects on dorsal horn neurones evoked by chemical stimulation at identified pressor and depressor sites in the lateral and ventrolateral periaqueductal grey (PAG). Stimulating micropipettes were inserted stereotaxically into the lateral or ventrolateral PAG at sites where microinjection of dl-homocysteic acid (DLH) evoked increases or decreases respectively in mean arterial blood pressure. The effects of DLH microinjection at these sites were tested against the responses of dorsal horn neurones to noxious and innocuous stimuli applied to their cutaneous receptive fields. Single unit extracellular recordings were made from 15 Class 1 (low-threshold) and 37 Class 2 (wide dynamic range) dorsal horn neurones in laminae II–VI of the lower lumbar spinal cord. The responses of Class 1 neurones to innocuous prodding of their receptive fields were unaffected by neuronal activation in either the lateral or ventrolateral PAG. The nociceptive (noxious pinch/heat) responses of most Class 2 neurones were strongly inhibited by chemical stimulation in either sector of the PAG. The low threshold (prod) responses of the same neurones were generally unaffected or only weakly inhibited by identical stimulation, regardless of stimulation site. No significant differences were found between the effects of lateral vs. ventrolateral PAG stimulation on the responses of dorsal horn neurones. These results do not support the view that dorsal horn neurones may be inhibited with different selectivities by hyper- and hypotensive regions of the PAG.

Fictive vocalization in the cat.

A new animal model is described for the study of fictive vocalization, evoked by electrical stimulation of the periaqueductal grey (PAG) in decerebrate, paralyzed and artificially ventilated cats. Changes in activities of the phrenic, abdominal, recurrent and superior laryngeal nerves induced by PAG stimulation after induction of paralysis were compared with the activities of these nerves or the muscles innervated by them during actual vocalization induced before paralysis. The PAG-induced respiratory and laryngeal motor pattern in paralyzed cats is similar to that of PAG-induced vocalization. Furthermore, the switch from inspiration to vocal-like activity is usually synchronized with lung inflation in both paradigms. It is concluded that fictive vocalization can be induced by PAG stimulation in paralyzed cats.

Central origins of substance P-like immunoreactive fibers and terminals in the spinal trigeminal caudal subnucleus in the rat

After trigeminal rhizotomy, some substance P-like immunoreactive (SP-LI) fibers and terminals in the spinal trigeminal caudal subnucleus (Vc), specially in its superficial laminae (laminae I and II), still remained in the rat. Employing a combination of Fluoro-Gold retrograde tracing and immunofluorescence histochemical staining for SP, we found that the main central origins of these SP-LI fibers and terminals were midbrain periaqueductal gray (PAG), nucleus raphe magnus (NRM) and other raphe nuclei, and nucleus reticularis gigantocellularis pars α; all of them are important structures of the endogenous pain control system. The present results provided morphological evidence for PAG or NRM stimulation could inhibit neuronal activities in the Vc evoked by orofacial nociceptive stimulation and also suggested that SP might be an important neurotransmitter or neuromodulator for endogenous pain control system.

Role of GABA receptor subtypes in inhibition of primate spinothalamic tract neurons: difference between spinal and periaqueductal gray inhibition.

1. gamma-Aminobutyric acid (GABA) is thought to inhibit both pre- and postsynaptically the transfer of nociceptive signals from primary afferent fibers to spinal dorsal horn sensory cells, including spinothalamic tract (STT) neurons. The inhibition can be mediated by both GABAA and GABAB receptors. We now attempt to characterize the synaptic inhibition of STT cells by spinal GABAA and GABAB receptors in anesthetized monkeys and to analyze the roles of these two receptor subtypes in the inhibition of STT cellular activity produced by stimulation in the periaqueductal gray (PAG). 2. Iontophoretic release of GABA or muscimol (a selective GABAA receptor agonist) onto STT cells elicited a profound and dose-related inhibition of the responses of all cells tested to noxious cutaneous stimuli. Only four cells (16.7%) were found to be inhibited when baclofen (a selective GABAB receptor agonist) was applied iontophoretically. However, a strong and dose-dependent inhibition of the responses to cutaneous mechanical and thermal stimuli was obtained in all cells examined when baclofen was administered into the dorsal horn through a microdialysis fiber. The inhibitory effects were mainly on nociceptive inputs. 3. The inhibition of cellular activity by GABAA and GABAB agonists could be selectively antagonized by specific antagonists applied through a microdialysis fiber. 4. The excitatory responses evoked by pulsed release of glutamic acid (GLUT) were also inhibited in a dose-related manner by iontophoretic application of GABA and muscimol, but not by baclofen. A high dose of baclofen administered by microdialysis resulted in only a small decrease in GLUT-evoked excitatory responses. 5. Infusion of GABAA and GABAB antagonists into the dorsal horn by microdialysis caused an increase in both background activity and responses to cutaneous stimuli, suggesting that there is a tonic GABAergic inhibition of STT cells. 6. The inhibition of responses to mechanical and thermal stimulation of the cutaneous excitatory receptive field resulting from stimulation in PAG was significantly antagonized in most of the STT cells tested when the GABAA antagonist bicuculline was infused into the spinal dorsal horn through a microdialysis fiber. In contrast, the inhibition produced by PAG stimulation in most of the cells examined was not significantly antagonized by the GABAB antagonists phaclofen or 3-amino-propyl(diethoxymethyl)phophinic acid (CGP35348) administered into the spinal dorsal horn by microdialysis. 7. Our results support the contention that GABAergic mechanisms in the spinal dorsal horn normally exert a tonic modulation of nociceptive inputs through both GABAA and GABAB receptors. The evidence provided here indicates that GABAA receptors located on primate STT neurons contribute to a postsynaptic inhibitory effect on the transmission of peripheral nociceptive inputs. A possible presynaptic GABAA action was not investigated. Our finding of a GABAB-receptor-mediated inhibition is consistent with the view that both pre- and postsynaptic GABAB receptors are involved in inhibitory modulation of spinal nociceptive transmission. Finally, it is suggested from this study that primate spinal GABAA, but not GABAB receptors, are involved in mediating the descending inhibition induced by PAG stimulation.

Chapter 20 A concept for the final common pathway of vocalization and lordosis behavior in the cat

Publisher Summary Periaqueductal gray matter (PAG) integrates the autonomic and somatic components of survival and mating behavior. For example, PAG stimulation has been demonstrated to produce simultaneous changes in nociception and blood pressure, vocalization, jumping, arching of the back, micturition, and lordosis. This chapter discusses the projections from the PAG to the nucleus retroambiguus (NRA), especially the projections from the NRA to motoneurons not so much in the framework of vocalization but in the framework of lordosis behavior. NRA neurons also project directly to motoneurons, bypassing the spinal interneurons. The NRA neurons play the role of premotor interneurons, similar to the ones in the spinal intermediate zone. Premotor interneurons usually receive peripheral Ia afferents or input from cortico- and rubrospinal tracts, but this is not the case for the premotor interneurons in the NRA. A probable explanation is that the NRA interneurons have a special function that is, activating a set of motoneuronal cell groups located throughout the lower brainstem and entire spinal cord, totally independent of voluntary input or spinal reflexes. In this context, the NRA receives afferents from other sources such as the PAG and respiration related brainstem nuclei.

Ventrolateral orbital cortex and periaqueductal gray stimulation-induced effects on on- and off-cells in the rostral ventromedial medulla in the rat.

On- and off-cells of the rostral ventromedial medulla are thought to be involved in bulbospinal inhibition of ascending nociceptive information. Experiments were carried out in lightly anaesthetized rats to assess the effects of prefrontal cortex stimulation on the responses of neurons in the rostral ventromedial medulla. For comparison purposes, effects of periaqueductal gray stimulation were also investigated. Single unit activity was recorded in the rostral ventromedial medulla and on-, off- and neutral-cells were identified based on the tail nocifensor reflex to noxious heat. Short (0.1-1 s) and long (10-15 s) trains of bipolar electrical stimulation (100-300 Hz) were delivered to the ventrolateral orbital cortex of the rat forebrain and the periaqueductal gray. Short-train stimulation of the periaqueductal gray (including dorsolateral, ventrolateral and the dorsal raphé regions) excited 58% (25 of 43) of on-cells and 44% (seven of 16) of off-cells in the rostral ventromedial medulla. Long trains blocked the noxious stimulus-evoked pause of all seven off-cells tested and blocked the excitatory response of two, and enhanced one of three on-cells. Such stimulation also inhibited or abolished the tail-flick reflex at currents below 100 microA. Glutamate microinjections into the periaqueductal gray inhibited the noxious-evoked response of two off- and two on-cells and increased the tail-flick latency. Short-train stimulation of the ventrolateral orbital cortex (100-400 microA) excited eight of 25 on-cells and inhibited the ongoing activity of 10 of 14 off-cells. Long-train ventrolateral orbital cortex stimulation (5-15 s, 100-200 microA, 200-300 Hz) enhanced the noxious evoked responses of 10 of 11 on-cells, prolonged the noxious heat-evoked pause of all of four off-cells and decreased the tail-flick latency (pronociception). The results of this study support the proposed role of on- and off-cells in descending inhibition of nociception from the periaqueductal gray and implicate the ventrolateral orbital cortex in the control of this pathway.

Brain stem integration of vocalization: role of the nucleus retroambigualis.

1. The descending pathways that mediate the periaqueductal gray (PAG)-evoked coordination of respiratory, laryngeal, and orofacial activity for vocalization have yet to be delineated. Two hypotheses have been offered. One theory is that this activity is mediated by a diffuse descending projection to parvocellular reticular interneurons, adjacent to the relevant laryngeal and orofacial motoneuronal pools. The second hypothesis is that the motor activity for vocalization is integrated via a projection from the PAG to a caudal medullary column of neurons, the nucleus retroambigualis (NRA). These hypotheses were tested with the use of a series of medullary transections combined with PAG stimulation. Transections that eliminated, in a series of caudal-to-rostral steps, the NRA, also eliminated the PAG-evoked cricothyroid and most of the thyroarytenoid laryngeal motor activity. These results indicate that the final common pathway for much of the laryngeal activity in PAG-evoked vocalization includes un initial synapse in the caudal medulla, presumably in the NRA. 2. The electromyographic changes evoked by microinjection of D,L-homocysteic acid (DLH) in the NRA of the unanesthetized, precollicular decerebrate cat were analyzed in order to delineate the NRA contribution to the coordinated respiratory, laryngeal, and oral muscle changes in vocalization. A total of 415 DLH injection sites were located at or caudal to the level of the obex. Vocalization was evoked at 46 of these sites, which were all confined to a restricted region of the ventrolateral medulla 1-3 mm caudal to the obex. This region corresponded to the rostral half of the NRA and the immediately adjacent medullary tegmentum. 3. In all experiments evidence was obtained that variable muscle activation, rather than functional and integrated muscle patterns, was represented within the NRA. Vocalization evoked by DLH microinjection in the NRA was usually associated with excitation of the cricothyroid, thyroarytenoid, external oblique, internal oblique, internal intercostal, and diaphragm muscles that occurred in a different manner from site to site. That is, injection at sites separated by 0.3-0.5 mm evoked quite different responses. 4. NRA-evoked vocalization was compared with PAG-evoked vocalization using small injections (1.5-4.5 nl) into each region. As well, larger microinjections (15-120 nl) into NRA were made for comparison with previous results from the PAG using similar doses. Within the PAG, stereotyped and relatively "fixed" patterns of muscle activity are represented, whereas within the NRA there was no representation of specific muscle patterns, but rather a partial topographic separation of "premotor neurons" regulating different muscles. Correspondingly, stereotyped vocalizations were never evoked from the NRA. Further, most NRA-evoked vocalizations were unusual in quality and would not be identified generally as feline. 5. Evidence was obtained for a separation of pathways from the PAG regulating sound production and orofacial modulation of that sound. In contrast to the results from the PAG, excitation of NRA neurons rarely evoked activity in the oral muscles (genioglossus or anterior belly of digastric) or orofacial modulation of sound production. 6. Our finding suggests that the NRA serves as an important substrate for the generation of respiratory pressure and larynges adduction, which are two essential aspects of not only vocalization but also several behaviors involving Valsava maneuvers such as coughing, vomiting, and defecation.

Release into ventriculo-cisternal perfusate of β-endorphin- and met-enkephalin-immunoreactivity: effects of electrical stimulation in the arcuate nucleus and periaqueductal gray of the rat

To examine the resting and evoked release of the endogenous opioid peptides β-endorphin and Met-enkephalin from brain, we examined the levels of the respective immunoreactivities in the lateral ventricle-cisterna magna perfusate of the halothane-anesthetized rat. Ten Hz but not 100 Hz stimulation in the arcuate nucleus (ARC) of the hypothalamus released β-endorphin immunoreactivity (β-EPir) to the perfusate, whereas 100 Hz but not 10 Hz stimulation in the periaqueductal gray (PAG) of the mid brain released Met-enkephalin immunoreactivity (MEir). MEir was not released by stimulation in ARC and β-EPir was not released by stimulation in PAG. Characterization of the released β-EPir and MEir by high performance liquid chromatography showed that authentic β-endorphin and Met-enkephalin were the major constituents of β-EPir and MEir, respectively. Systemic administration of the dopaminergic antagonist haloperidol increased plasma, but not perfusate levels of β-EPir. Both the opioid antogonist naloxone and the NMDA antagonist MK-801 failed to affect β-EPir or MEir release. ARC and PAG stimulation inhibited a nociceptive reflex (tail-dip in 52.5°C water), and naloxone did not reliably reverse this inhibition. These data support the previously suggested possibility of opioid mediation of stimulation induced analgesia, although we were unable to confirm the theory by naloxone reversibility in this study. Furthermore, the data support the assumption that measurement of opioid peptides in cerebrospinal fluid is a relevant approach in research aimed at elucidating the physiological and pathophysiological roles of endogenous opioid peptides.

Midbrain suppression of limb withdrawal and tail flick reflexes in the rat: correlates with descending inhibition of sacral spinal neurons.

1. The aim of this study was to utilize new quantitative behavioral methods in rats to investigate the effects of electrical stimulation in midbrain analgesia areas on the magnitude of flexion hindlimb withdrawal and tail flick reflexes evoked by graded noxious heating. Electrophysiological experiments were then done with the use of these animals to correlate behavioral data with the effects of identical midbrain stimulation on sacral dorsal horn neuronal responses to graded heating of the tail. 2. To quantify limb withdrawals, electromyographs (EMGs) were recorded in biceps femoris during withdrawals elicited by noxious heat stimuli (40-52 degrees C, 5 s, 2-min intervals) delivered to the plantar surface of the hind paw, without and during concomitant electrical stimulation (100 ms, 100-Hz trains, 3/s, 10-600 microA) in midbrain periaqueductal gray (PAG) or laterally adjacent reticular formation (LRF) via previously implanted electrodes. The same animals were tested with the use of a tail flick paradigm modified to allow measurement of the force of tail movements in three orthogonal planes and thereby calculate the overall force vector of tail flicks elicited by graded noxious radiant heat pulses (38-58 degrees C, 5 s, 2-min intervals) delivered to the tail, again with and without concomitant PAG or LRF stimulation. Finally, the same rats were anesthetized with pentobarbital sodium and microelectrode recordings made from single sacral dorsal horn neurons responsive to the same noxious heat stimuli delivered to the tail to assess effects of PAG and LRF stimulation. 3. PAG and LRF stimulation suppressed the magnitude of limb flexor EMGs, and tail flick force vectors, in an intensity-dependent manner. Recruitment of suppression of both limb withdrawal EMGs and tail flicks was generally more effective for LRF compared with PAG stimulation, although mean thresholds for suppression were similar. Tail flick force and limb withdrawal EMGs recorded from the same rat in separate sessions were suppressed about equally in a majority of cases. 4. Limb withdrawal EMG magnitude increased monotonically from threshold (approximately 40 degrees C) to 52 degrees C. The population stimulus-response function was fit equally well by linear regression or a 2 degrees polynomial function (r2 = 0.79 for both). PAG stimulation significantly reduced the mean slope of the stimulus-response function (to 73%; n = 15), whereas LRF stimulation shifted it toward the right with a smaller slope reduction (to 85%) and 3 degrees C increase in threshold.(ABSTRACT TRUNCATED AT 400 WORDS)

Electrophysiological analysis of midbrain periaqueductal gray influence on cardiovascular neurons in the ventrolateral medulla oblongata

Stimulation of sites in the rostral or caudoventral periaqueductal gray (PAG) results in substantial increases in mean blood pressure (MBP) and heart rate (HR). The efferent pathways from these PAG subregions possibly include a relay in the ventrolateral medulla oblongata (VLM), where neurons involved in maintaining vasomotor tone are located. Extracellular recordings were made from 21 cardiovascular neurons in the rostral VLM (RVLM) and from 6 cardiovascular neurons in the caudal VLM (CVLM) of the rat. These neurons showed barosensitivity and cardiac rhythmicity. In addition, the activity of 54 noncardiovascular and nonrespiratory units was recorded. Responses to electrical stimulation of sites in the (rostral or caudal) PAG were studied in 16 of the 21 cardiovascular RVLM neurons, the 6 CVLM neurons, and 46 of the 54 noncardiovascular neurons. Eight of the RVLM neurons were excited by rostral PAG stimulation; the poststimulus time histograms showed a constant latency in five units (32 ± 3 ms). This suggests the presence of relatively direct (although not monosynaptic) excitatory pathways from the rostral PAG to cardiovascular neurons in the RVLM, consisting of slowly conducting fibers (0.2-0.3 m/s). Five RVLM neurons did not respond to rostral PAG stimulation. Three units were tested with caudal PAG stimulation: one was excited, one inhibited, and one was unresponsive. The six cardiovascular CVLM neurons did not respond to PAG stimulation. Of the 46 noncardiovascular neurons, 14 cells were excited, 7 inhibited, and 2 cells antidromically activated. These results confirm earlier findings, extending them to the rostral PAG. They supply further evidence for the influence of the PAG on the cardiovascular function-related neuronal circuitry in the VLM.

Gastric distension modulates hypothalamic neurons via a sympathetic afferent path through the mesencephalic periaqueductal gray

The effects of gastric distension on extracellulary recorded single neuronal activity in the lateral hypothalamuslateral preoptic area-medial forebrain bundle (LPA-LH-MFB), other areas of the hypothalamus, mesencephalic periaqueductal gray (PAG), and other areas associated with the mesencephalic reticular formation were determined in the anesthetized rat Gastric distension was produced by filling a gastric balloon with water using a calibrated infusion pump. Experimental conditions were based on previous studies that simulated gastric distension during fluid consumption In the rat. The effects of stomach distension using water at body temperature and room temperature were compared. Neurons in both the hypothalamus and mesencephalon were modulated by gastric distension. Hypothalamic neurone exhibited responses associated with gastric distension and exhibited interactions between distension and temperature stimulation of the stomach. Neurons in the mesencephalic periaqueductal gray (PAG) and associated reticular formation also were modulated by these gastric stimuli. When the PAG was electrically stimulated, similar responses to gastric distension and PAG stimulation were observed for hypothalamic neurons. The effects of gastric distension on hypothalamic neurons were reduced or eliminated when the PAG stimulating electrode site was destroyed by electrocoagulation. In addition, the microiontophoretic application of horseradish peroxidase at hypothalamic neuronal recording sites where gastric distension effects were observed resulted in the retrograde labeling of neurons in the PAG. These gastric stimulation-induced effects on hypothalamic and mesencephallc neuronal activity were attenuated but were not permanently eliminated by bilateral cervical vagotomy. However, these effects were significantly reduced or eliminated by bilateral transection of the cervical sympathetic chain or spinal transection at the first cervical level. Because the filling of balloons placed into the abdominal cavity close to the stomach had no similar effects on neural activity, these results can be attributed primarily to the activation of gastric mechano- and temperature-sensitive receptors. These results indicate that the effects of gastric temperature/distension stimulation under these conditions are mediated to a large degree by sympathetic afferents. The PAG is clearly involved as one of the mesencephalic relays for gastric afferent input to the hypothalamus.

Differential inhibitory mechanisms in VPL versus intralaminar nociceptive neurons of the cat: I. Effects of periaqueductal gray stimulation.

Nociceptive thalamic units receiving afferent input from the greater splanchnic nerve (SPL) were recorded from the nucleus ventralis posterolatealis (VPL) and intralaminar nuclei in urethane-chloralose anesthetized cats. The effects of stimulating the periaqueductal gray (PAG), or the nucleus raphe dorsalis (NRD) on responses of nociceptive thalamic units were investigated. Forty-eight nociceptive specific (NS) and 20 wide dynamic range (WDR) units with SPL input were found in the shell region of the caudal VPL. Following electrical stimulation of either the ventral PAG or the NRD, responses to SPL input were inhibited in all NS and WDR units tested. Responses of these units to electrical stimulation of spinothalamic tract fibers in the ventrolateral funiculus (VLF) were also inhibited following the PAG/NRD stimulation. These results suggest that PAG/NRD stimulation-produced inhibition of both NS and WDR units may be partially mediated by an ascending antinociceptive mechanism. Intralaminar nociceptive units with SPL input were found in the nuclei centralis lateralis (CL), paracentralis (Pc), and parafascicularis (Pf). The effects of conditioning electrical stimulation of either the ventral PAG or the NRD on responses of intralaminar nociceptive units were studied. Of 113 intralaminar nociceptive units studied, 68 units were unaffected, 23 units were excited and 22 units were inhibited following the conditioning stimulation. In the units in which responses to SPL stimulation were inhibited, responses elicited by electrical stimulation of the mesencephalic reticular formation (MRF) were also inhibited. These data suggest that although there is an ascending inhibitory pathway from PAG/NRD to intralaminar nuclei, this system is far less potent compared with the ascending inhibitory system acting upon the VPL.

Descending modulation of central neural plasticity in the formalin pain test

Subcutaneous injection of formalin produces a biphasic profile of pain response: a transient early phase followed by a tonic late phase. A number of studies have indicated that the development of the late phase of formalin pain is dependent upon prolonged changes in central neural function produced by neural activity that is generated during the early phase (i.e. central sensitization). In support of this, the present study demonstrates that stimulation- or morphine-produced analgesia derived from the periaqueductal grey (PAG) during the early phase prevents the developement of the late phase. these results suggest that descending mechanisms of pain inhibition, as reflected by PAG stimulation- and morphine-produced analgesia, can prevent the development of central neural plasticity following injury.

Electrical Stimulation of the Dorsal Midbrain Periaqueductal Gray Suppresses Peripheral Blood Natural Killer Cell Activity

This study examined the effects of dorsal midbrain periaqueductal gray (PAG) stimulation on both splenic and peripheral blood natural killer (NK) cell function as well as the proliferative response of lymphocytes to phytohemagglutinin mitogen. Male Sprague-Dawley rats were implanted with bipolar electrodes in the dorsal PAG. Following recovery, bipolar electrical stimulation eliciting a flight response was delivered at the rate of one/min for 30 min to freely moving rats. While dorsal PAG stimulation did not alter mitogen response or splenic NK activity, stimulation of this region of the PAG produced a marked decrease in peripheral blood NK response. In order to begin to explore a possible mechanism regulating suppression of peripheral blood NK activity, naltrexone (10 mg/kg) was administered prior to dorsal PAG stimulation. The results of this experiment replicated the findings that demonstrated suppression of peripheral blood NK following dorsal PAG stimulation. Naltrexone did not effect PAG induced suppression of peripheral blood NK. These findings point to the importance of the dorsal aspect of the PAG in the regulation of peripheral blood NK activity and further suggest that this phenomenon may not be opioid mediated.

Glycine and GABA A antagonists reduce the inhibition of primate spinothalamic tract neurons produced by stimulation in periaqueductal gray

Amino acids are demonstrated to be important neurotransmitters mediating the inhibitory transmission from nucleus raphe magnus to spinal nociceptive dorsal horn neurons. In this study, the role of glycine and GABA in the inhibitory processes evoked by stimulation in periaqueductal gray (PAG) of responses of primate spinothalamic tract (STT) neurons to cutaneous mechanical and thermal stimuli was investigated by examining the effects of strychnine and bicuculline, antagonists of glycine and GABA A receptors, respectively, introduced into the dorsal horn through a microdialysis fiber. The inhibitory effects of iontophoretic application of glycine and GABA A agonists on STT cell activity evoked by noxious mechanical stimulation of the skin were selectively blocked by their specific antagonist, strychnine or bicuculline, infused into the dorsal horn. Similarly, intra-spinal application of strychnine or bicuculline resulted in a significant reduction in the PAG stimulation-induced inhibition of responses of STT cells to cutaneous stimuli. This reduction was mainly on the PAG-induced inhibition of the responses to noxious mechanical stimuli. Our results suggest that glycinergic and GABAergic inhibitory interneurons in the spinal cord dorsal horn synapsing on STT cells are activated during stimulation in PAG and contribute to descending antinociceptive actions.