GABA(A) receptor regulation of kyphotic nursing and female sexual behavior in the caudal ventrolateral periaqueductal gray of postpartum rats

Bilateral lesions of the ventrolateral caudal periaqueductal gray inhibit lordosis and kyphosis, the postures of female sexual receptivity and maternal nursing that are characterized respectively by dorsoflexion and ventroflexion of the spinal column. These lesions also inhibit the solicitation behaviors that accompany lordosis, but they do not impair retrieval or licking of pups. We tested the hypothesis that reproductive behaviors affected by these lesions are tonically inhibited by activity of the GABA(A) receptor via site-specific manipulations of receptor activity. Rats were bilaterally implanted during pregnancy with guide cannulae aimed at the caudal periaqueductal gray and ovariectomized on day 1 postpartum. Microinfusions (0.25 microl/side) of saline or drug took place on days 5 and 7 postpartum into the dorsolateral column and on days 9 and 11 into the ventrolateral column. Five minutes post-infusion dams were reunited with their pups and their maternal behavior was observed for 30 min. Feminine sexual behaviors were evaluated post-weaning after another set of microinfusions in the ventrolateral caudal periaqueductal gray. Potential facilitation of kyphosis and lordosis was tested with the GABA(A) antagonist bicuculline (15 ng/side) during sub-threshold conditions, i.e., non-suckling pups or sub-threshold ovarian hormone dosages; potential inhibition of these postures was tested with the GABA(A) agonist muscimol (125 ng/side) during optimal conditions, i.e., suckling pups or supra-threshold ovarian hormone treatments. Dorsolateral drug manipulations were ineffective. In the ventrolateral periaqueductal gray bicuculline significantly increased and muscimol significantly decreased kyphosis, lordosis, and sexual solicitations compared with saline. Retrieval and licking of pups were not altered by GABA(A) manipulations. These findings suggest that the reproductive postures of female rats, lordosis and kyphosis, as well as sexual solicitations, are tonically inhibited by the neurotransmitter GABA within the ventrolateral caudal periaqueductal gray in the midbrain. In contrast, retrieval and licking of pups appear to be under separate neurochemical or neuroanatomical control, or both. Further, this tonic inhibition is likely relieved by excitatory somatosensory inputs to this site, from mounting and suckling respectively.     

Oxytocin receptor activity in the ventrocaudal periaqueductal gray modulates anxiety-related behavior in postpartum rats

Postpartum rats are less anxious than diestrous virgin females, a phenomenon requiring that mothers have recent contact with their infants. Oxytocin (OT) is one of many neurochemicals released intracerebrally while mothers interact with infants, and we investigated whether OT receptor activity in the ventrocaudal periaqueductal gray (cPAGv) contributes to mothers' reduced anxiety. Infusion of the highly specific OT receptor antagonist, desGly-NH2,d(CH2)5[D-Tyr2,Thr-sup-4]OVT, into the cPAGv reduced the percentage of time dams spent in the open arms of an elevated plus-maze, but had no effect on the open-arm behavior of diestrous virgins. Conversely, after separating dams from their litters for 4 hr to increase anxiety, a lower (2 ng) but not higher (5 ng) dose of OT infused into each hemisphere of the cPAGv doubled the percentage of time dams spent in open arms, but did not do so in virgins. OTergic manipulations inconsistently affected risk-assessment behaviors (stretch-attend postures, head dips) in both virgins and dams. Therefore, OT receptor activation in the cPAGv is an important consequence of contact with infants that reduces some anxiety-related behaviors in mother rats.     

Effects of induced thyroid deficiency on the development of suckling behavior in rats

The effects of propylthiouracil (PTU) on the ontogeny of suckling behavior in rats were examined. The drug was given at two dosage levels of 0.3% and 0.5% respectively mixed with rat diet throughout gestation and suckling. The thyroid glands of treated fetuses and pups and of untreated control animals of the same age groups were monitored by histologic examination. At the behavioral level, the frequency of individual movements of head, forelimbs and mouth was significantly reduced in treated fetuses. The combination movements of head, mouth and forelimbs showed severe deficits both quantitatively and qualitatively for all ages in the experimental group from day 18 of gestation. Hypothyroid pups of dams raised on 0.5% PTU were unable to attach to the nipple of the mother and died within a few days. Pups of dams raised on 0.3% PTU showed longer latencies for nipple attachment, and their gross motor movements of rooting and suckling were greatly impaired. These results have been discussed in relation to the development of suckling behavior to indicate that, during ontogeny, some decisive step in the integration of individual movements takes place in utero from day 18 of gestation. This coincides with the establishment of pituitary thyroid relationship, which is continued through postnatal stages.     

Effects of persistent nociception on periaqueductal gray glycine release

Glycine is a candidate nociception inhibitory transmitter in specific brain regions, like for example the spinal cord, the thalamic nuclei and the periaqueductal gray matter. However, quantitative changes in glycine released in these brain regions during peripheral inflammation episodes have not been characterized in awake animals. To address this issue, an in vivo microdialysis study was carried out in freely moving rats in order to analyse periaqueductal gray matter extracellular glycine concentration following unilateral formalin injection into the dorsal skin of the right hind-paw. The extracellular concentration of glutamine was also evaluated in order to analyse whether or not a non-neurotransmitter amino acid was equally modified. Intra-periaqueductal gray matter tetrodotoxin perfusion reduced extracellular glycine concentration (-44+/-5%), but did not change the glutamine dialysate values. Peripheral injection of formalin reduced the glycine release during the early phase (-62+/-8%) and the late phase (-36+/-6%) of hyperalgesia, although not during the analgesic period. Perfusion with naloxone (300microM) neither prevented the formalin-induced decreases in extacellular glycine concentration, nor modified the perfusate basal values of glycine and glutamine.These results show that, contrary to what has been recognized on the interactive role of opioids and GABA into the periaqueductal gray matter (i.e. opioid disinhibition), endogenous opioids seem not to modulate the activity of glycinergic neurons in the same midbrain area. In the light of these preliminary data, it is reasonable to suppose that GABA and glycine are probably not co-released at the level of periaqueductal gray matter of the rat.     

Sex differences in locomotor activity following beta-endorphin in the ventrolateral periaqueductal gray

Both morphine and selective opioid receptor agonists produce typical biphasic effects upon activity in male rats with initial reductions in activity followed by subsequent increases in activity. Sex differences have been observed in a number of opioid-mediated responses, including the observation that microinjection of the opioid peptide, beta-endorphin into the ventrolateral periaqueductal gray (vlPAG) produces more pronounced analgesia in male rats relative to female rats. The present study evaluated whether beta-endorphin (5.2-26 microg) in the vlPAG produced biphasic activity effects in male and female rats. Both male and female rats displayed initial reductions in total, ambulatory, and stereotypic activity following beta-endorphin in the vlPAG. Whereas male rats displayed subsequent (90-120 min) increases in total and ambulatory activity following beta-endorphin in the vlpAG, female rats tested during the estrous stage of their estrous cycle failed to display any changes. These sex differences in the opioid modulation in the pattern of activity measures in the vlPAG are discussed in terms of the roles of sex hormones in opioid peptide processing within the vlPAG.     

大鼠中脑导水管周围灰质向下丘脑orexin神经元的纤维投射

目的:明确中脑导水管周围灰质(PAG)神经元调节下丘脑orexin神经元的解剖学基础。方法:首先采用逆行示踪技术,将霍乱毒素B亚单位(CTb)作为逆行示踪剂注射到下丘脑orexin神经元胞体分布的区域,免疫组织化学方法染色观察PAG内CTb逆行标记细胞的分布特点;接着采用顺行示踪技术和免疫组化相结合的方法,将生物素化葡聚糖胺(BDA)作为顺行示踪剂注射到PAG,观察下丘脑内BDA标记的神经纤维与orexin神经元的重叠分布特点;最后运用免疫电镜技术观察BDA标记轴突终末与orexin神经元之间的突触联系。结果:CTb注射到单侧下丘脑orexin神经元区域后,在PAG观察到大量CTb标记神经元,注射点同侧标记细胞数明显占优,其中PAG外侧区(lPAG)和腹外侧区(vlPAG)可见大量CTb逆标神经元,背外侧区(dlPAG)和背内侧区(dm PAG)可见少量标记神经元;注射点对侧lPAG及vlPAG区域也观察到少量CTb标记神经元。BDA注射到单侧lPAG后,下丘脑内双侧均可观察到BDA标记的神经纤维,但注射点同侧标记神经纤维数量明显占优。BDA标记纤维与orexin神经元胞体在穹窿背侧接近不定带(ZI)的区域(本文中将此区域称作"穹窿上区")形成显著的重叠分布。透射电镜下观察到BDA标记的轴突终末与orexin神经元之间形成突触联系,且多为非对称性类型。结论:lPAG投射到下丘脑的神经纤维在穹窿上区与orexin神经元形成优势重叠分布、并与orexin神经元的胞体或树突形成以非对称性为主的突触联系,提示orexin神经元可能在lPAG的直接支配下发挥其调节觉醒状态、摄食行为等功能。     

Distinct regions of periaqueductal gray (PAG) are involved in freezing behavior in hooded PVG rats on the cat-freezing test apparatus

The periaqueductal gray (PAG) is considered to be an exit relay for defensive responses. Studies have shown that the ventrolateral periaqueductal gray (vlPAG) plays a role in the expression of freezing behavior whereas dorsolateral periaqueductal gray (dlPAG) is involved on both freezing and active forms of defensive behaviors. To further elucidate this theory, lesioned vlPAG and dlPAG rats were exposed to a cat in the cat-freezing test apparatus. Subsequently, a 7-day repeated exposure to a cat was done on the vlPAG and dlPAG lesioned rats. Results showed that the vlPAG lesioned rats demonstrated significant decrease in freezing behavior and corresponding increase in locomotor activity, while the dlPAG lesioned rats failed to show any significance. Subsequent repeated exposure of the vlPAG lesioned rats to a cat for 7 days showed a gradual decrease in freezing behavior with significance shown at days 5, 6 and 7 while the dlPAG lesioned rats failed to show any changes. These results suggest that vlPAG regulates freezing behavior in hooded PVG rats.     

Hypovolemic shock: critical involvement of a projection from the ventrolateral periaqueductal gray to the caudal midline medulla

Previous research has suggested that the ventrolateral column of the periaqueductal gray (vlPAG) plays a crucial role in triggering a decompensatory response (sympathoinhibition, hypotension, bradycardia) to severe blood loss. vlPAG excitation triggers also quiescence, decreased vigilance and decreased reactivity, the behavioral response which usually accompanies hypovolemic shock. The aim of this study was to identify, in unanesthetized rats, the main descending pathway(s) via which vlPAG neurons trigger sympathoinhibition and bradycardia in response to severe blood loss. Firstly, immediate early gene (c-Fos) expression was used to identify vlPAG neurons selectively activated by severe blood loss. Subsequently, the specific medullary projections of these vlPAG neurons were defined by combined c-Fos, retrograde tracing (double-label) experiments. It was found that vlPAG neurons selectively activated by severe hemorrhage project overwhelmingly to the vasodepressor portion of the caudal midline medulla (CMM). Previous studies indicate that this CMM region mediates behaviorally-coupled cardiovascular adjustments and the findings described here fit with the idea that CMM neurons are uniquely recruited by salient challenges, the adaptive responses to which require more than reflexive homeostatic cardiovascular adjustments.     

Fluoxetine attenuates the effects of pentylenetetrazol on rat freezing behavior and c-Fos expression in the dorsomedial periaqueductal gray

The aim of the study was to investigate the role of the periaqueductal gray (PAG) in anxiolytic-like actions of fluoxetine in animals treated with an anxiogenic drug, pentylenetetrazol (PTZ), and subjected to fear conditioning procedure. The data showed that PTZ given at the dose of 30 mg/kg 15 min before a retention trial significantly decreased freezing reaction (p<0.01), and potently enhanced rat locomotor activity (p<0.01), in comparison to the control group. These effects were reversed by prior (60 min) administration of fluoxetine (20 mg/kg). Simultaneously, PTZ significantly increased c-Fos expression in the dorsomedial periaqueductal gray (DMPAG), examined 2h after the retention trial, in comparison to the control group (p<0.01). Fluoxetine (20 mg/kg) administered 60 min before PTZ reversed this effect. PTZ given at the same dose and time interval in the open field test did not affect rat locomotor behavior. Importantly, fluoxetine pretreatment did not change PTZ concentration in brain tissue. Our experiment based on PTZ-enhanced aversive conditioning revealed that acutely administered fluoxetine antagonized PTZ-induced panic-like behavior, and this phenomenon was accompanied by inhibition of activity of DMPAG.     

Buspirone induced acute and chronic changes of neural activation in the periaqueductal gray of rats

5-HT_1A modulation within the midbrain periaqueductal gray (PAG) is closely associated with anxiety- or panic-like behavior. Several findings have demonstrated that the properties of buspirone (a 5-HT_1A partial agonist) would function as either anxiolytic or panicolytic in both clinical and laboratory animal research. In this study, we have investigated the neuronal activity occurring within the different regions of the PAG induced by buspirone treatment. Twenty-eight albino Wistar rats (350–400 g) were injected with either acute or chronic saline/buspirone (each, n=7), respectively. Our results show that buspirone treatment reduced locomotor activity, body weight and fecal boli, particularly in the chronic buspirone group. Two-way ANOVA revealed a significant decrease of c-Fos-immunoreactive (ir) cells expression in all regions of the rostral PAG after both acute and chronic buspirone (acute buspirone (AB) and chronic buspirone (CB), respectively) treatment. However, no effects on c-Fos-ir were detected in the caudal lateral periaqueductal gray (lPAG) and ventrolateral periaqueductal gray (vlPAG) in both the AB and CB groups, and in the dorsolateral periaqueductal gray (dlPAG) of the CB group. Interestingly, c-Fos-ir cells in the dorsomedial periaqueductal gray (dmPAG) column were reduced consistently in both the rostral and caudal PAG in both AB and CB groups. Besides, in all regions the number of c-Fos-ir cells was higher in the AB than in the CB group with exception of the rostral lPAG. In conclusion, the main anxiolytic effect of buspirone was specifically localized in all regions of the rostral PAG and in the caudal dmPAG. However, the caudal dlPAG, lPAG and vlPAG were found to be ineffective to buspirone treatment, probably due to their distinctive function in mediating higher level of anxiety in defensive behavior. This indicates that the longitudinal anatomical structure of the PAG possesses a different level of receptor sensitivity of 5-HT_1A in the pathophysiology of anxiety and panic disorder.     

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.     

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 curent information about the anatomy, physiology, and pharmacology of the PAG, a model is proposed to account for the interactions between these different functional components.     

Neuronal activity in the periaqueductal gray and bordering structures during vocal communication in the squirrel monkey

In seven freely moving squirrel monkeys (Saimiri sciureus), the neuronal activity in the periaqueductal gray (PAG) and bordering structures was registered during vocal communication, using a telemetric single-unit recording technique. In 9.3% of the PAG neurons, a vocalization-correlated activity was found. Four reaction types could be distinguished: a) neurons, showing an activity burst immediately before vocalization onset; b) neurons, firing during vocalization, and starting shortly before vocalization onset; c) neurons, firing exclusively during vocalization; d) neurons, firing in the interval between perceived vocalizations (i.e. vocalizations produced by group mates) and self-produced vocal response. All PAG neurons showed a marked vocalization-type specificity. None of the neurons reflected simple acoustic parameters, such as fundamental frequency or amplitude, in its discharge rate. None of the neurons reacted to vocalizations of other animals not responded to by the experimental animal. All four reaction types found in the PAG were also found in the reticular formation bordering the PAG, though in lower density.     

Maternal behaviour in lactating rats stimulates c-fos in glutamate decarboxylase-synthesizing neurons of the medial preoptic area, ventral bed nucleus of the stria terminalis, and ventrocaudal periaqueductal gray

Increased activity of the immediate-early gene c-fos can be observed in many areas of the lactating rat brain after dams physically interact with pups and display maternal behaviour. These sites include the medial preoptic area, ventral bed nucleus of the stria terminalis, and the ventrolateral caudal periaqueductal gray, each of which is critical for the normal performance of particular maternal behaviours. The phenotype of cells in these areas that show increased c-fos activity after maternal behaviour, however, is unknown. Via double-label immunocytochemistry, we determined if the population of cells in these sites that express c-fos after maternal behaviour in lactating rats overlaps with the population that expresses the 67,000 mol. wt isoform of glutamate decarboxlyase, the synthesizing enzyme for the inhibitory neurotransmitter GABA. Lactating rats were separated from pups beginning on day 5 postpartum, and 48h later half were allowed to interact with a litter of pups for 60min whereas the other half were not. Dams re-exposed to pups were highly maternal, retrieving and licking them as well as displaying prolonged nursing behaviour that included milk letdown. Both groups of dams had a similar number of 67,000 mol. wt glutamate decarboxylase-immunoreactive cells in each site, although the number of 67,000 mol. wt glutamate decarboxylase-immunoreactive cells per microscopic field was significantly greater in the caudal ventrolateral periaqueductal gray than in the ventral bed nucleus of the stria terminalis, which in turn was greater than the medial preoptic area. In pup-stimulated dams, two to fourfold more Fos-immunoreactive cells were found in these three sites compared with non-stimulated controls. Labeling for Fos immunoreactivity and 67,000 mol. wt glutamate decarboxylase immunoreactivity was heterogeneous within each site. In the medial preoptic area, more Fos-immunoreactive and 67,000 mol. wt glutamate decarboxylase-immunoreactive cells (either single or dual-labeled) were found dorsally than ventrally. In the ventral bed nucleus of the stria terminalis, more Fos-immunoreactive and 67,000 mol. wt glutamate decarboxylase-immunoreactive cells were found medially than laterally. Within the caudal ventrolateral periaqueductal gray, 67,000 mol. wt glutamate decarboxylase-immunoreactive labeling was greatest ventromedially, while high numbers of Fos-immunoreactive nuclei were found both ventromedially and ventrolaterally. In pup-stimulated dams, more than half (53% in the medial preoptic area, 59% in the ventral bed nucleus of the stria terminalis, and 61% in the caudal ventrolateral periaqueductal gray) of the total population of Fos-immunoreactive cells also expressed 67,000 mol. wt glutamate decarboxylase.These results suggest that many of the neurons in these sites that show elevated c-fos activity after maternal behaviour are either local inhibitory interneurons or provide inhibitory input to other neural sites. These inhibitory mechanisms may be critical for the display of postpartum nurturance, possibly facilitating maternal behaviour by removing tonic inhibition on sites necessary for maternal responding or by restricting activity in neural sites that inhibit it.     

Afferent projections to the periaqueductal gray in the rabbit

The afferents to the periaqueductal gray in the rabbit have been described following hydraulic pressure injection of horseradish peroxidase at various sites throughout this structure. Every third section was reacted with tetramethylbenzidine, for the localization of afferent neurons. At the site of the deposit alternate sections were reacted with tetramethylbenzidine, Hanker-Yates reagent, or diaminobenzidine, for comparative assessment of the injection site. A large number of retrogradely labelled cells, assessed by bright- and dark-field microscopy, were observed in a wide range of areas throughout the brain. Major labelled areas within the telencephalon were cortical areas 5, 20, 21, 32 and 40. Within the diencephalon, the hypothalamus contained quantitatively by far the largest number of labelled cells. Of these nuclei, the dorsal pre-mammillary nucleus contained the largest number of labelled cells. Considerable labelling was also found within medial and lateral preoptic nuclei, anterior hypothalamic area, and ventromedial hypothalamic nucleus. Another diencephalic region containing a significant number of retrogradely labelled neurons was the zona incerta. At midbrain, pontine and medullary levels, additional labelled regions were: the substantia nigra, cuneiform nucleus, parabigeminal nucleus, raphe magnus, and reticular areas. Heavy labelling was seen within the periaqueductal gray itself, rostral and caudal to deposits placed within each subdivision. In addition, a large number of other areas labelled throughout the brain (Tables 2A-D). Not only were some differences noted in the pattern of labelled cells with deposits placed rostrally or caudally within periaqueductal gray, but certain topographical differences with respect to the degree of labelling within nuclei were also seen with injection sites ventral, lateral or dorsal to the aqueduct. In addition, a further difference was noted, in that over one third of the areas labelled with deposits in just one or other of the "divisions" within periaqueductal gray. The results therefore suggest that the periaqueductal gray might be divisible to some extent on the basis of connectivity with intrinsic subdivisions of the complex. It is hoped that, with time, it might prove possible to resolve any such differential input in functional terms. The wide variety of afferent input to the periaqueductal gray, and its strategic location, would seem to place it in a unique position for integrating and modifying a diversity of motor, autonomic, hormonal, sensory and limbic influences.(ABSTRACT TRUNCATED AT 400 WORDS)     

Contrasting effects of nitric oxide and corticotropin-releasing factor within the dorsal periaqueductal gray on defensive behavior and nociception in mice

The anxiogenic and antinociceptive effects produced by glutamate N-methyl-D-aspartate receptor activation within the dorsal periaqueductal gray (dPAG) matter have been related to nitric oxide (NO) production, since injection of NO synthase (NOS) inhibitors reverses these effects. dPAG corticotropin-releasing factor receptor (CRFr) activation also induces anxiety-like behavior and antinociception, which, in turn, are selectively blocked by local infusion of the CRF type 1 receptor (CRFr1) antagonist, NBI 27914 [5-chloro-4-(N-(cyclopropyl)methyl-N-propylamino)-2-methyl-6-(2,4,6-trichlorophenyl)aminopyridine]. Here, we determined whether i) the blockade of the dPAG by CRFr1 attenuates the anxiogenic/antinociceptive effects induced by local infusion of the NO donor, NOC-9 [6-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-hexanamine], and ii) the anxiogenic/antinociceptive effects induced by intra-dPAG CRF are prevented by local infusion of N^ω-propyl-L-arginine (NPLA), a neuronal NOS inhibitor, in mice. Male Swiss mice (12 weeks old, 25-35 g, N = 8-14/group) were stereotaxically implanted with a 7-mm cannula aimed at the dPAG. Intra-dPAG NOC-9 (75 nmol) produced defensive-like behavior (jumping and running) and antinociception (assessed by the formalin test). Both effects were reversed by prior local infusion of NBI 27914 (2 nmol). Conversely, intra-dPAG NPLA (0.4 nmol) did not modify the anxiogenic/antinociceptive effects of CRF (150 pmol). These results suggest that CRFr1 plays an important role in the defensive behavior and antinociception produced by NO within the dPAG. In contrast, the anxiogenic and antinociceptive effects produced by intra-dPAG CRF are not related to NO synthesis in this limbic midbrain structure.     

Lidocaine blockade of amygdala output in fear-conditioned rats reduces Fos expression in the ventrolateral periaqueductal gray

We showed recently that conditioned fear to context induces Fos expression in the ventrolateral periaqueductal gray [Neuroscience (1997) 78, 165-177]. Neurons in this region are thought to play an important role in the expression of freezing during conditioned fear. To test the possibility that this activation comes directly from the amygdala, we looked at changes in Fos expression after a unilateral blockade of the ventral amygdalofugal pathway with lidocaine. The pathway contains fibres originating from the central nucleus of the amygdala that project directly and mainly ipsilaterally to the ventrolateral periaqueductal gray. Conditioned fear was evoked by re-exposing rats to the same box in which they had previously received electric footshocks. The test re-exposure was preceded by a unilateral microinjection of lidocaine (2%, 0.5-1 microl; n = 20) or saline (n = 14). Lidocaine was also tested in non-conditioned animals (n = 13). The results show that, when lidocaine was microinjected in the medial part of the central nucleus of the amygdala or along the ventral amygdalofugal pathway of conditioned rats, fear-induced Fos expression in the ventrolateral periaqueductal gray was reduced on the side ipsilateral to the injection (up to 37% reduction in comparison to the contralateral side). Ipsilateral reductions were also observed with saline, but they were weaker (maximum of 27% reduction). Fos expression remained low on both sides in the non-fear-conditioned animals injected with lidocaine. Finally, although freezing was only partly reduced in the conditioned animals unilaterally injected with lidocaine, it was significantly correlated to the ipsilateral reduction in Fos expression. This study provides direct evidence that the projection from the central nucleus of the amygdala to the ventrolateral periaqueductal gray is activated during fear and that it contributes to the Fos response of the ventrolateral periaqueductal gray.     

Activation of mu opioid receptors in the ventrolateral periaqueductal gray inhibits reflex micturition in anesthetized rats

This study tested the hypothesis that morphine and other opiates cause urinary retention by activating mu opioid receptors in the midbrain periaqueductal gray (PAG) region. Selective mu, delta and kappa receptor agonists were microinjected into the PAG of urethane-anesthetized rats and the amplitude and incidence of bladder contractions were recorded during continuous saline infusion. Arterial pressure was monitored through a femoral artery catheter. Microinjection of the mu receptor agonist DAMGO into the ventrolateral PAG (vlPAG) suppressed volume-evoked bladder contractions completely. Bladder contractions ceased within 5 min of DAMGO injection and remained essentially undetectable for the rest of the 20 min recording period. Microinjection of the delta receptor agonist DPDPE into the vlPAG did not significantly affect either the amplitude of bladder contractions or the time interval separating contractions. The kappa receptor agonist U-69593 caused no discernible change in amplitude but increased the interval between bladder contractions significantly. Microinjection of DAMGO, DPDPE or U-69593 into the lateral or dorsolateral PAG columns was ineffective. DAMGO injection into the vlPAG increased arterial pressure whereas DPDPE and U-69593 produced a small but significant depressor response. These data support the hypothesis that mu and kappa receptors in the vlPAG participate in the micturition reflex.     

Lidocaine blockade of amygdala output in fear-conditioned rats reduces Fos expression in the ventrolateral periaqueductal gray

We showed recently that conditioned fear to context induces Fos expression in the ventrolateral periaqueductal gray [Neuroscience (1997) 78, 165–177]. Neurons in this region are thought to play an important role in the expression of freezing during conditioned fear. To test the possibility that this activation comes directly from the amygdala, we looked at changes in Fos expression after a unilateral blockade of the ventral amygdalofugal pathway with lidocaine. The pathway contains fibres originating from the central nucleus of the amygdala that project directly and mainly ipsilaterally to the ventrolateral periaqueductal gray. Conditioned fear was evoked by re-exposing rats to the same box in which they had previously received electric footshocks. The test re-exposure was preceded by a unilateral microinjection of lidocaine (2%, 0.5–1 μl; n=20) or saline (n=14). Lidocaine was also tested in non-conditioned animals (n=13). The results show that, when lidocaine was microinjected in the medial part of the central nucleus of the amygdala or along the ventral amygdalofugal pathway of conditioned rats, fear-induced Fos expression in the ventrolateral periaqueductal gray was reduced on the side ipsilateral to the injection (up to 37% reduction in comparison to the contralateral side). Ipsilateral reductions were also observed with saline, but they were weaker (maximum of 27% reduction). Fos expression remained low on both sides in the non-fear-conditioned animals injected with lidocaine. Finally, although freezing was only partly reduced in the conditioned animals unilaterally injected with lidocaine, it was significantly correlated to the ipsilateral reduction in Fos expression.This study provides direct evidence that the projection from the central nucleus of the amygdala to the ventrolateral periaqueductal gray is activated during fear and that it contributes to the Fos response of the ventrolateral periaqueductal gray.