Stimulation of the rat subthalamic nucleus is neuroprotective following significant nigral dopamine neuron loss.

Deep brain stimulation of the subthalamic nucleus (STN-DBS) is efficacious in treating the motor symptoms of Parkinson's disease (PD). However, the impact of STN-DBS on the progression of PD is unknown. Previous preclinical studies have demonstrated that STN-DBS can attenuate the degeneration of a relatively intact nigrostriatal system from dopamine (DA)-depleting neurotoxins. The present study examined whether STN-DBS can provide neuroprotection in the face of prior significant nigral DA neuron loss similar to PD patients at the time of diagnosis. STN-DBS between 2 and 4 weeks after intrastriatal 6-hydroxydopamine (6-OHDA) provided significant sparing of DA neurons in the SN of rats. This effect was not due to inadvertent lesioning of the STN and was dependent upon proper electrode placement. Since STN-DBS appears to have significant neuroprotective properties, initiation of STN-DBS earlier in the course of PD may provide added neuroprotective benefits in addition to its ability to provide symptomatic relief.

Functional characteristics of the midbrain periaqueductal gray.

The major functions of the midbrain periaqueductal gray (PAG), including pain and analgesia, fear and anxiety, vocalization, lordosis and cardiovascular control are considered in this review article. The PAG is an important site in ascending pain transmission. It receives afferents from nociceptive neurons in the spinal cord and sends projections to thalamic nuclei that process nociception. The PAG is also a major component of a descending pain inhibitory system. Activation of this system inhibits nociceptive neurons in the dorsal horn of the sinal cord. The dorsal PAG is a major site for processing of fear and anxiety. It interacts with the amygdala and its lesion alters fear and anxiety produced by stimulation of amygdala. Stimulation of PAG produces vocalization and its lesion produces mutism. The firing of many cells within the PAG correlates with vocalization. The PAG is a major site for lordosis and this role of PAG is mediated by a pathway connecting the medial preoptic with the PAG. The cardiovascular controlling network within the PAG are organized in columns. The dorsal column is involved in pressor and the ventrolateral column mediates depressor responses. The major intrinsic circuit within the PAG is a tonically-active GABAergic network and inhibition of this network is an important mechanism for activation of outputs of the PAG. The various functions of the PAG are interrelated and there is a significant interaction between different functional components of the PAG. Using the current information about the anatomy, physiology, and pharmacology of the PAG, a model is proposed to account for the interactions between these different functional components.

Connections between the central nucleus of the amygdala and the midbrain periaqueductal gray: topography and reciprocity.

Previous reports indicate that the midbrain periaqueductal gray and the central nucleus of the amygdala are interconnected but the organization of these projections has not been characterized. We have analyzed this reciprocal circuitry using anterograde and retrograde tracing methods and image analysis. Our findings reveal that innervation of periaqueductal gray from the central nucleus of the amygdala is extensive and discretely organized along the rostrocaudal axis of periaqueductal gray. In addition, the reciprocal projection from periaqueductal gray to the central nucleus of the amygdala is more extensive and more highly organized than previously suggested. Multiple or single discrete injections of wheatgerm agglutinin-horseradish peroxidase into several rostrocaudal levels of periaqueductal gray retrogradely labeled a substantial population of neurons, predominantly located in the medial division of the central nucleus of the amygdala. Tracer injections into the central nucleus revealed a high degree of spatial organization in the projection from this nucleus to periaqueductal gray. Two discrete longitudinally directed columns in dorsomedial and lateral/ventrolateral periaqueductal gray are heavily targeted by central amygdalar inputs throughout the rostral one-half to two-thirds of periaqueductal gray. Beginning at the level of dorsal raphe and continuing caudally, inputs from the central nucleus terminate more uniformly throughout the ventral half of periaqueductal gray. In addition, a substantial population of periaqueductal gray neurons were retrogradely labeled from the central nucleus of the amygdala; these were heterogeneously distributed along the rostrocaudal axis of periaqueductal gray, and included both raphe and non-raphe neurons. Thus, the present study demonstrates that periaqueductal gray receives heavy, highly organized projections from the central nucleus of the amygdala and, in turn, has reciprocal connections with the central nucleus. Previous studies have demonstrated that longitudinally organized columns of output neurons located in dorsomedial and lateral/ventrolateral periaqueductal gray project to the ventral medulla. Thus, there may be considerable overlap between the two longitudinally organized terminal input columns from the central nucleus of the amygdala and the two longitudinal columns of descending projection neurons from periaqueductal gray to the ventral medulla. The central nucleus of the amygdala has been implicated in a variety of emotional/cognitive functions ranging from fear and orienting responses, defensive and aversive reactions, associative conditioning, cardiovascular regulation, and antinociception. Many of these same functions are strongly represented in the periaqueductal gray. It is noteworthy that the present results demonstrate that lateral periaqueductal gray, a preeminent central trigger site for behavioral and autonomic components of the defense/aversion response, is heavily targeted by inputs from the central nucleus of the amygdala at all levels of periaqueductal gray.(ABSTRACT TRUNCATED AT 400 WORDS)

Directionally specific changes in arterial pressure induce differential patterns of fos expression in discrete areas of the rat brainstem: a double-labeling study for Fos and catecholamines.

Although the nucleus tractus solitarii (NTS) has been established as the primary site of synaptic integration for the baroreceptor reflex, the higher-order pathways responsive to, and mediating, changes in vasomotor tone are not well characterized. We used immunohistochemistry to determine the distribution of cells expressing the Fos protein following pharmacologically induced, directionally specific changes in arterial pressure. The goal of this investigation was to determine if this immediate early gene product is differentially expressed in neurons of the rat brainstem following increased (pressor) versus decreased (depressor) arterial blood pressure (AP). Because brainstem catecholaminergic (CA) cell groups have been implicated in cardiovascular regulation, a double-labeling immunohistochemical procedure was used to examine the distribution of Fos in CA cells. Animals received continuous intravenous infusion of either a vasoconstrictor (l-phenylephrine hydrochloride), a vasodilator (sodium nitroprusside), or physiological saline. Extensive Fos-like immunoreactivity (FLI) was induced in both the pressor and depressor conditions in the NTS, caudal ventrolateral medulla (CVLM), rostral ventrolateral medulla (RVLM), A5, locus coeruleus (LC), Kolliker-Fuse, and parabrachial nucleus (PBN). These regions have all been implicated in central cardiovascular regulation. There were differences in the anatomical distribution of Fos-positive cells along the rostrocaudal axis of CVLM in the pressor and depressor conditions. Specifically, increased AP induced significantly more FLI cells within the rostral aspects of CVLM, whereas decreased AP resulted in a significantly greater number of FLI cells within the caudal CVLM. This result suggests that selective vasomotor responses differentially engaged discrete subsets of neurons within this brainstem region. Overall, approximately 50% of CA-immunoreactive cells were also FLI (CA-FLI) in the A1, A5, and A7 regions. Interestingly, increased AP produced significantly more CA-FLI double-labeled cells within the caudal than rostral A1 compared with depressor and control groups. Additionally, increased AP yielded significantly less CA-FLI double-labeled cells within the caudal A2 region. This suggests that CA barosensitive neurons in the CVLM/A1 and NTS/A2 regions are functionally segregated along the rostrocaudal axis of these structures. While twice as many PNMT-FLI double-labeled neurons were found in the C1-C3 regions following vasomotor changes versus saline control, there were no differences in the numbers or anatomical locations of labeled cells between pressor versus depressor groups. The results of this study indicate that (1) tonic changes in AP induce robust Fos expression in brainstem cardiovascular areas and (2) neurons responsive to specific directional changes in arterial pressure are segregated in some brainstem regions.

Fos expression induced by changes in arterial pressure is localized in distinct, longitudinally organized columns of neurons in the rat midbrain periaqueductal gray.

The distribution of neurons expressing Fos within the periaqueductal gray (PAG) following pharmacologically induced high or low blood pressure was examined to determine (1) if PAG neurons are responsive to changes in arterial pressure (AP) and (2) the relationship of these cells to the functionally defined hypertensive and hypotensive columns in PAG. Changes in AP differentially induced robust Fos expression in neurons confined to discrete, longitudinally organized columns within PAG. Increased AP produced extensive Fos-like immunoreactivity within the lateral PAG, beginning at the level of the oculomotor nucleus. At the level of the dorsal raphe, Fos expression induced by increased AP shifted dorsally, into the dorsolateral division of PAG; this pattern of Fos labeling was maintained throughout the caudal one-third of PAG. Double-labeling for Fos and nicotinamide adenine dinucleotide phosphate diaphorase confirmed that Fos-positive cells induced by increased AP were located in the dorsolateral division of PAG at these caudal levels. Fos positive cells were codistributed, but not colocalized, with nicotinamide adenine dinucleotide phosphate diaphorase-positive cells. Decreased AP evoked a completely different pattern of Fos expression. Fos-positive cells were predominantly located within the ventrolateral PAG region, extending from the level of the trochlear nucleus through the level of the caudal dorsal raphe. Double-labeling studies for Fos and serotonin indicated that only 1-2 double-labeled cells per section were present. Saline infusion resulted in very few Fos-like immunoreactive cells, indicating that volume receptor activation does not account for Fos expression in PAG evoked by changes in AP. These results indicate that (1) substantial numbers of PAG neurons are excited by pharmacologically induced changes in AP and (2) excitatory barosensitive PAG neurons are anatomically segregated based on their responsiveness to a specific directional change in AP.

Differential expression of annexins I-VI in the rat dorsal root ganglia and spinal cord.

The annexins are a family of Ca(2+)-dependent phospholipid-binding proteins. In the present study, the spatial expression patterns of annexins I-VI were evaluated in the rat dorsal root ganglia (DRG) and spinal cord (SC) by using indirect immunofluorescence. Annexin I is expressed in small sensory neurons of the DRG, by most neurons of the SC, and by ependymal cells lining the central canal. Annexin II is expressed by most sensory neurons of the DRG but is primarily expressed in the SC by glial cells. Annexin III is expressed by most sensory neurons, regardless of size, by endothelial cells lining the blood vessels, and by the perineurium. In the SC, annexin III is primarily expressed by astrocytes. In the DRG and the SC, annexin IV is primarily expressed by glial cells and at lower levels by neurons. In the DRG, annexin V is expressed in relatively high concentrations in small sensory neurons in contrast to the SC, where it is expressed mainly by ependymal cells and by small-diameter axons located in the superficial laminae of the dorsal horn areas. Annexin VI is differentially expressed by sensory neurons of the DRG, being more concentrated in small neurons. In the SC, annexin VI has the most striking distribution. It is concentrated subjacent to the plasma membrane of motor neurons and their processes. The differential localization pattern of annexins in cells of the SC and DRG could reflect their individual biological roles in Ca(2+)-signal transduction within the central nervous system.

Preoptic projections to Barrington's nucleus and the pericoerulear region: architecture and terminal organization.

The medial preoptic area (MPO), a sexually dimorphic region, plays a pivotal role in neuroendocrine function and reproductive behavior. We recently reported that MPO projects heavily to the midbrain periaqueductal gray (PAG). We also noted that MPO projects to the dorsolateral pontine tegmentum. Here we identified the cells of origin of the MPO-->tegmental projection and delineated the terminal organization of MPO projections to Barrington's nucleus, locus coeruleus (LC), and the rostromedial pericoerulear region (pLCrm). Correlative cyto- and chemoarchitectonic studies were done to define better the nuclear groups of the dorsolateral pontine tegmentum. Retrograde tracing revealed that MPO neurons projecting to the dorsolateral pontine tegmentum are preferentially distributed in distinct subregions of MPO, including the sexually dimorphic medial preoptic nucleus (MPN). Anterograde tracing with wheat germ agglutinin-horseradish peroxidase or Phaseolus vulgaris leucoagglutinin demonstrated considerable target specificity in projections from MPO to the dorsolateral pontine tegmentum. Barrington's nucleus receives a dense focal input along its entire rostrocaudal axis. In addition, pLCrm is heavily targeted by MPO inputs; pLCrm contains a concentrated plexus of extranuclear dendrites of LC neurons. The lateral dorsal tegmental (LDT) nucleus and LC proper receive only sparse input from MPO. MPO projections to Barrington's nucleus could regulate micturition reflexes during reproductive behavior. The MPO-->pLCrm projection could influence noradrenergic LC neurons in relation to reproductive and/or gonadal steroid function. Given the strong established connections from olfactory structures to MPO, it is possible that the MPO-->LC pathway provides an anatomical substrate for olfactory modulation of arousal.

Susceptibility of feline spinal cord energy metabolism to severe incomplete ischemia.

Feline spinal cords were subjected to 10 to 30 minutes of severe incomplete ischemia (average reduction in blood flow of 92%) with and without 90 minutes of recirculation, and the L-2 segment was analyzed for high-energy phosphates and certain glycolytic metabolites. Spinal cord tissue lactic acid levels were stepwise elevated, and adenosine triphosphate (ATP), phosphocreatine (P-creatine), and glucose were progressively consumed by increasing durations of ischemia. However, upon restoration of blood flow, there was extensive recovery of energy metabolites and normalized lactic acid, demonstrating resumption of mitochondrial oxidative metabolism. These data indicate that the spinal cord can tolerate at least 30 minutes of severely reduced blood flow before recovery of energy metabolism is significantly impaired upon restitution of blood flow.

The effect of [Met]enkephalin on the periaqueductal gray neurons of the rat: an in vitro study.

Intracellular and extracellular recording techniques and in vitro preparation were used to examine the effect of [Met]enkephalin on the rat periaqueductal neurons. In the 20 cells that were recorded intracellularly, [Met]enkephalin caused an increase in the resting membrane conductance, hyperpolarization of the cell membrane, an increase in the firing threshold and a decrease in the spontaneous firing rate. This effect of [Met]enkephalin could be blocked by naloxone. The effect of [Met]enkephalin on 99 neurons was also examined using extracellular recording. In 59% of cells, pressure application of [Met]enkephalin caused a dose-dependent inhibition that could be blocked by naloxone; 15% of the cells were excited and the remaining neurons (26%) did not respond. Nineteen per cent of responsive cells were located in the dorsolateral subdivision; 41% in the ventrolateral and 13% in the dorsal regions. In 10 cells, perfusion with physiological saline solution/Mg did not alter the inhibitory effect of [Met]enkephalin. However, perfusion with physiological saline solution/Mg abolished the excitatory response to [Met]enkephalin in four cells. It is concluded that: (1) the major effect of [Met]enkephalin on periaqueductal gray cells is inhibition that occurs through a direct postsynaptic process. This inhibition is probably due to an increase in permeability to potassium; (2) a small population of periaqueductal gray cells are excited by [Met]enkephalin, probably through a presynaptic process.

The effects of diphenhydramine and SR142948A on periaqueductal gray neurons and on the interactions between the medial preoptic nucleus and the periaqueductal gray.

The midbrain periaqueductal gray contains both neurotensin type-1 and type-2 receptors. Behavioral studies have shown that the analgesic effect of neurotensin is mediated through its interaction with the type-2 receptors. These receptors specifically bind the type-1 histamine antagonist, levocabastine. Recently, it has been shown that another histamine-1 antagonist, diphenhydramine, blocks the analgesic effect of neurotensin. In addition, it has been shown that a non-peptide neurotensin antagonist, SR142948A, binds to both types of neurotensin receptors and blocks the analgesic effect of exogenously applied neurotensin. Major afferents to the periaqueductal gray arise from the medial preoptic nucleus of the hypothalamus. This region contains neurotensinergic neurons, and the expression of neurotensin mRNA in this region increases following cold-water swim stress that leads to opioid-independent analgesia. The goal of this study was to determine whether the responses of periaqueductal gray neurons to stimulation of the medial preoptic nucleus are modified by local injection of diphenhydramine and SR142948A. Because the cellular basis of the effects of diphenhydramine on periaqueductal gray neurons had not been reported, we also examined the effects of diphenhydramine on the baseline-firing rate and synaptic transmission using in vivo and in vitro methods. The results of the in vitro studies indicate that diphenhydramine concentrations above 500 nM significantly reduce the baseline firing of the periaqueductal gray neurons without a significant effect on the frequency of postsynaptic potentials. At concentrations below 100 nM, diphenhydramine has little effect on the baseline-firing rate but partially blocks the response to neurotensin. The results of the in vivo studies showed similar effects of diphenhydramine. At high concentrations it inhibited periaqueductal gray neurons, but at low concentrations it had no effect on the baseline-firing rate and it blocked the response to neurotensin and to medial preoptic nucleus stimulation. Unlike diphenhydramine, SR142948A had virtually no effect on the baseline-firing rate but blocked the response to neurotensin and to stimulation of the medial preoptic nucleus. It is concluded that: (1) SR142948A, at a dose that completely blocks the effect of exogenously applied neurotensin on periaqueductal gray neurons, has little effect on their baseline-firing rates. (2) Because of its effect on the baseline-firing rate, only low doses of diphenhydramine can be used as an antagonist of the neurotensin analgesic effect. (3) Responses of periaqueductal gray neurons to medial preoptic nucleus stimulation is, in part, mediated by a neurotensinergic network within the periaqueductal gray.

Physiological influence of lateral proisocortex on the midbrain periaqueductal gray: evidence for a role of an excitatory amino acid in synaptic activation.

Recent anatomical studies in this laboratory have demonstrated that the proisocortex cortex adjacent and dorsal to the rhinal sulcus is one of the major forebrain afferent inputs to the midbrain periaqueductal gray matter in the rat. The physiological influence(s) of this projection has not been examined. The present studies investigated the responses of periaqueductal gray neurons to chemical and electrical stimulation of proisocortex in chloral hydrate-anesthetized rats. In addition, the role of glutamate as a possible transmitter in excitatory proisocortex-periaqueductal gray synaptic responses was tested. Microinjection of D,L-homocysteate into proisocortex excited 44% (19/43), inhibited 37% (16/43) and had no effect on 19% of periaqueductal gray cells. The onset of D,L-homocystic acid-evoked responses ranged from 2 to 60 s; the duration of responses ranged from 1 to 18 min. Low-frequency, single-pulse electrical stimulation of proisocortex robustly altered neuronal discharge in 25% of periaqueductal gray neurons sampled; 10% (74/724) of neurons were excited and 15% (107/724) were inhibited. Insular cortex-evoked excitatory responses had a mean onset latency of 19.5 +/- 4.2 ms and a mean duration of 38.5 +/- 26.9 ms. Inhibitory responses had a mean onset latency of 26.2 +/- 15.6 ms and mean duration of 108.0 +/- 84.9 ms. Trains of high-frequency electrical stimulation of proisocortex excited 22% (13/59) and inhibited 25% (15/59) of periaqueductal gray cells tested. In separate experiments, stimulation electrodes were placed in periaqueductal gray to antidromically activate proisocortex neurons that project to periaqueductal gray.(ABSTRACT TRUNCATED AT 250 WORDS)

Cold water swim stress increases the expression of neurotensin mRNA in the lateral hypothalamus and medial preoptic regions of the rat brain.

Stress-induced analgesia is a well-documented phenomenon that occurs in all mammalian species. Forced cold water swim produces a type of stress-induced analgesia that is independent of mu opioid receptors. The neuropeptide neurotensin (NT) has been implicated in mu opioid-independent analgesia (MOIA), but the circuitry of this system is largely unknown. The medial preoptic area (MPO) and lateral hypothalamus (LH) are two regions that are known to modulate pain processing. These two regions also contain neurotensinergic projections to the periaqueductal gray, a region that has been shown to produce MOIA upon injection of NT. The goal of this study was to determine if cold water swim (CWS) stress, which produces MOIA, activates the NT-ergic systems in these two regions. In situ hybridization results indicate that CWS increases the level of NT mRNA within neurons in the MPO and LH, suggesting that these two regions are activated during this process.

BMY 7378: Partial agonist at spinal cord 5-HT(1A) receptors.

Recent data indicate that BMY 7378 demonstrates high affinity, selectivity and low intrinsic activity at hippocampal 5-HT(1A) receptors, suggesting that BMY 7378 may represent the first selective 5-HT(1A) functional antagonist. The present study examined the agonist and antagonist properties of BMY 7378 at spinal cord 5-HT(1A) receptors. In electrophysiological studies, iontophoretic administration of either the 5-HT(1A) agonist 8-OH-DPAT (43.8 +/- 5.4 nA) or BMY 7378 (46.3 +/- 5.2 nA) significantly inhibited the firing rate of wide-dynamic-range dorsal horn units indicating that BMY 7378 demonstrates significant intrinsic activity at spinal cord 5-HT(1A) receptors. Concomitant BMY 7378 and 8-OH-DPAT administration identified no BMY 7378 ejection current (20-100 nA) which antagonized the 8-OH-DPAT induced inhibition of dorsal horn unit activity. In behavioral studies in the spinal rat, 8-OH-DPAT increased the animals' sensitivity to noxious levels of mechanical stimulation (ED(50) = 269 +/- 24 nmol/kg) as did BMY 7378 (ED(50) = 295 +/- 70 nmol/kg) with no statistically significant difference in the maximal response (Y(max)) observed. Concomitant BMY 7378 and 8-OH-DPAT administration identified no BMY 7378 dose (10-1100 nmol/kg) which blocked the hyperalgesic effect of 8-OH-DPAT, rather, each drug produced similar effects which were additive. Further, the 5-HT(1A) agonist effects of BMY 7378 were blocked by the 5-HT(1A) antagonist, spiperone. Therefore, both the electrophysiologic and reflex data indicate that BMY 7378 possesses significant intrinsic activity at spinal cord 5-HT(1A) receptors, and like 8-OH-DPAT is a partial agonist at these receptors. Differences in BMY 7378 intrinsic activity at spinal cord as opposed to hippocampal 5-HT(1A) receptors are discussed in terms of regional differences in G-proteins coupled to 5-HT(1A) receptors in these two CNS regions.

Identification of a novel 5-HT(1) binding site in rat spinal cord.

High affinity, specific [(3)H]5-hydroxytryptamine (5-HT) binding to spinal cord synaptosomes was examined to identify the 5-HT receptor subtypes present. Computer nonlinear regression analysis of competition studies employing 8-OH-DPAT indicated that this 5-HT(1A) selective agonist demonstrated high affinity competition (K(i = 1.3 nM)) for 24.6 +/- 0.7% of the total [(3)H]5-HT binding sites. Competition studies employing the 5-HT(1B) selective agonist RU24969, in the presence of 100 nM 8-OH-DPAT, indicated that RU24969 demonstrated high affinity (K(i = 1.1 nM)) competitive inhibition for 26.2 +/- 1.4% of all [(3)H]5-HT binding sites. Neither 5-HT(1C), 5-HT(1D), 5-HT(2) nor 5-HT(3) selective compounds demonstrated any high affinity competition for the residual 49% of specific [(3)H]5-HT binding. Therefore, three major classes of [(3)H]5-HT binding sites could be demonstrated in spinal cord synaptosomes: 5-HT(1A), 5-HT(1B) and a novel [(3)H]5-HT binding site which respectively represented 25, 26 and 49% of spinal cord synaptosomal [(3)H]5-HT binding. Further studies focusing on the function of the latter binding site are needed to determine if the presently identified novel binding site is the major 5-HT(1) receptor subtype present in spinal cord.

Bulbospinal and intraspinal thyrotropin releasing hormone systems: modulation of spinal cord pain transmission.

Recent research indicates that pain processing by spinal nociceptive neurons is modulated by: 1) descending bulbospinal pathways originating in the rostral ventral medulla (RMV) and 2) short intraspinal peptide systems located in the dorsal horn. Immunohistochemical studies have identified both bulbospinal and intraspinal thyrotropin releasing hormone (TRH) systems, however, the role of these two TRH systems in pain modulation has not been defined. The purpose of the present study was to explore the role of the bulbospinal and intraspinal TRH systems in pain modulation. Three TRH mediated neural network were examined in electrophysiologic experiments; each model predicting specific testable outcomes. Model A represented a direct TRH projection from RVM to spinal cord. This model predicted that the effect of RVM stimulation and TRH micropressure administration on dorsal horn unit activity would be in the same direction, either both inhibitory effects or both excitatory. Of 44 dorsal horn units inhibited by RVM stimulation, 82% were excited by TRH, therefore, Model A was rejected. Model B interposed an intraspinal neuron between the bulbospinal TRH projection and the dorsal horn nociceptive unit. This model predicted that desensitization of spinal TRH receptor systems should block the effect of RVM stimulation on dorsal horn neuron. In TRH desensitization experiments, no attenuation or blockade of RVM stimulation was observed, therefore, the effect of RVM stimulation did not appear mediated by TRH and Model B was rejected.(ABSTRACT TRUNCATED AT 250 WORDS)

The role of acetylcholine in the function of the nucleus raphe magnus and in the interaction of this nucleus with the periaqueductal gray.

The nucleus raphe magnus (NRM) plays an important role in the inhibition of pain. Although this region receives afferents from several areas of the brain, the afferent input from the periaqueductal gray (PAG) has been shown to have significant physiological importance. Together, these two sites constitute the major component of a descending network involved in pain inhibition. In this study the role of acetylcholine (ACh) in the function of the NRM was investigated and the possibility that ACh may be a transmitter between the PAG and the NRM was tested. ACh was applied iontophoretically. Scopolamine and gallamine were used to test the type of cholinergic receptors that are present in the NRM. The results of this study shows the following. (1) The majority of the cells in the NRM are excited by ACh. (2) This response to ACh is partially or totally blocked by scopolamine whereas gallamine does not block the response. (3) There is no correlation between the excitatory response to stimulation of PAG and to ACh. There are cells that respond to PAG stimulation by inhibition but are excited by ACh and there are a few cells that are inhibited by ACh but are excited by PAG stimulation. (4) Scopolamine, at a dose that blocks the ACh response, does not block the response to PAG stimulation. (5) There is no correlation between the response to ACh and the type of projection (direct or indirect) to the spinal cord, as tested by stimulation of the dorsolateral funiculus. From these results it is concluded that ACh is an excitatory transmitter at the NRM region but this transmitter does not mediate the interaction between the PAG and NRM.

Electrophysiological characterization of the projection from the nucleus raphe magnus to the lateral reticular nucleus: possible role of an excitatory amino acid in synaptic activation.

Numerous studies have shown that the lateral reticular nucleus (LRN), located in the caudal ventrolateral medulla, is an important nuclear region in the descending analgesia system. Activation of this brainstem region, either electrically or chemically, results in a reduction in nociceptive threshold. In addition, destruction of LRN abolishes the tonic descending inhibition present on dorsal horn neurons. Recent neuroanatomical tracing studies have shown that the nucleus raphe magnus (NRM), long implicated in nociception, sends direct projections to LRN; however, no information exists regarding the physiological characteristics of this pathway, nor its role in the endogenous descending analgesia system. The purpose of this study was to physiologically characterize the synaptic influence(s) of projections from the NRM to the LRN using electrophysiological recording, electrical and chemical stimulation, and iontophoretic techniques. Sixty-one percent of LRN neurons responded to single pulse stimulation of NRM; 52% of the responsive cells were excited and 48% were inhibited. The mean latency to onset of excitation was 4.9 +/- 1.2 ms. High frequency (100 Hz) electrical stimulation of NRM influenced 69/102 neurons; 52% (36/69) were excited, while 48% (33/69) were inhibited. Microinjection of glutamate into NRM significantly modified the discharge of 83% (93/112) of LRN cells tested; of these, 71% were inhibited, while 29% were excited. In 35 cells the effects of the excitatory amino acid antagonist kynurenic acid (KYN) were studied. In 75% of the cells excited by glutamate administration into the NRM (18/24), KYN partially antagonized this response. In 11 LRN cells inhibited by NRM chemical stimulation, KYN had no effect on this inhibition. Overall, 95% of the LRN cells responsive to NRM stimulation were also responsive to noxious peripheral stimulation, indicating that these cells are receiving ascending information from the spinal cord regarding somatosensory stimulation as well as receiving descending input from the NRM. It is concluded that LRN neurons are highly responsive to both noxious peripheral stimulation and NRM efferent activation, and that this region plays a significant role as an integrator for both ascending and descending information.

Evidence that an excitatory connection between the periaqueductal gray and nucleus raphe magnus mediates stimulation produced analgesia.

Both electrical stimulation and injection of morphine into the midbrain periaqueductal gray (PAG) produce analgesia in the rat. There is evidence that this analgesic effect is mediated by a descending system that involves nucleus raphe magnus (NRM) and adjacent reticular formation. In the studies reported here, the activity of the cells in the PAG was increased by microinjection of glutamate in this area and its effect on both the activity of single cells in the NRM and on a flexion reflex elicited by noxious heat was measured. It is shown that an increase in the firing rate of the cells in the PAG is associated with a raised threshold for flexion and is also correlated with an increase in the firing rate of a majority of the cells in the NRM. This effect on the flexion reflex can be abolished by (a) lesion of the nucleus raphe magnus and a small area of the reticular formation surrounding this nucleus and (b) by nalazone 20 min after its i.v. injection. It is concluded that there is an excitatory connection between the periaqueductal gray and the nucleus raphe magnus and that activation of this system can cause analgesia.

Response of nucleus raphe magnus neurons to electrical stimulation of nucleus cuneiformis: role of acetylcholine.

There is considerable evidence that cells in the ventral medulla which includes nucleus raphe magnus (NRM) and nucleus magnocellularis are involved in a descending pain inhibitory system. Anatomical studies indicate a strong projection from nucleus cuneiformis (NCF) to the ventral medulla and histochemical studies suggest that many NCF neurons are cholinergic. Therefore, we investigated the effect of NCF stimulation on NRM unit activity and explored the possible role of acetylcholine (ACh) in this interaction. Of 180 NRM neurons examined, 43% were excited and 14% were inhibited by NCF stimulation. The average latency to the peak excitatory response was about 14 ms with a range of 5-32 ms. There was a tendency for the response latencies to cluster around 5 and 14 ms. Inhibitory responses were between 10 and 65 ms in duration. The anatomical specificity of the effective stimulation site was assessed by determining the response of a given NRM neuron to stimulation of areas dorsal and ventral as well as within NCF. The most reliable and intense responses of NRM neurons was observed with electrode placements within NCF. The most effective NCF region for activating NRM neurons corresponded to that region of NCF that contains a large population of neurons that project directly to NRM as seen in the present histochemical studies. The involvement of ACh in the interaction between NCF and NRM was examined with iontophoretic application of ACh and its antagonists. Of NRM neurons that responded to ACh, 79% were excited, an effect which was blocked by scopolamine.(ABSTRACT TRUNCATED AT 250 WORDS)

The medial preoptic nucleus of the hypothalamus modulates activity of nitric oxide sensitive neurons in the midbrain periaqueductal gray.

Stimulation of the medial preoptic nucleus of the hypothalamus (MPO) has been shown to produce decreases in mean arterial pressure (MAP) by a pathway involving the periaqueductal gray region of the midbrain (PAG). Previous studies have shown that the injection of nitric oxide (NO) donating compounds into the dorsal PAG also decreases MAP, while the injection of nitric oxide synthase (NOS) inhibitors increases MAP. Collectively these studies suggest that the MPO elicited hypotensive response may involve NO production in PAG neurons. In this study, we investigated this hypothesis. We found that: (1) Bilateral injection of the NOS inhibitor 7-nitro indazole (7-NI) into the dorsolateral PAG cell columns produced elevations in MAP in a highly consistent and site specific fashion. (2) Microinjection of 7-NI in quantities that were too low to directly influence MAP blocked the MPO evoked hypotensive response in 9/11 cases. (3) While 41% of dorsal PAG neurons had baseline firing rates that were sensitive to 7-NI, 69% of PAG neuronal responses to MPO stimulation were blocked by 7-NI. (4) Inhibitory responses that were not blocked by 7-NI had significantly shorter latencies to onset in the presence of 7-NI. (5) PAG neurons that projected to the medulla exhibited similar electrophysiologic response patterns. Our results suggest the following: (1) The dorsolateral PAG contains a NO producing hypotensive network. (2) The MPO elicited hypotensive response may utilize this network. (3) Stimulation of the MPO elicits NO dependent responses from PAG neurons, some of which do project to medullary-cardiovascular control centers.