The neuropeptide FLFQPQRFamide (NPFF) is a FRMRamide like peptide able to modulate morphine-induced analgesia (1). Recently, we showed the presence of both NPFF immunoreactivity and NPFF receptors in the dorsal horn superficial layers of the rat spinal cord which is the area where fine diameter nociceptive afferent fibers terminate (2, 3). Moreover, we showed that NPFF receptors are different from opiate receptors suggesting that morphine modulating activity of NPFF was supported by the activation of specific receptors (4). In the present study we address the question of the relationship between NPFF receptors and primary afferent fibers carrying nociceptive cutaneous and muscular informations from the fore- or hindlimb

The organization of the efferent projections from the pontine parabrachial (pPB) area to the amygdala has been studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L), a sensitive and selective anterograde axonal marker, into restricted subregions of the pPB area.The results confirmed that the pPB area primarily projected onto the ipsilateral nucleus centralis of the amygdala (Ce), and to a lesser extent onto the ipsilateral posterior basolateral (BLP), anterior basomedial (BMA), and amygdaloid cortical (ACo) nuclei of the amygdala. Substantial projections were also found in the substantia innominata dorsal/ventral portion of the globus pallidus (SId/GPv), substriatal (SStr), and fondus striatal (FStr) regions which continue the amygdala rostrally.The results demonstrated that the projections of the pPB area onto the Ce were topically organized: 1) The region of the pPB area mainly including the medial subnucleus (pPBm), the waist area (pPBwa), and a thin rostral lamina of the ventral lateral subnucleus (pPBvl) projects primarily to the medial portion of the Ce (CeM). Dense projections were also found in the BLP, BMA, and ACo nuclei of the amygdala, and in the SId/GPv, SStr, and FStr rostral areas. 2) The region of the pPB mainly including the rostral portion of the central lateral subnucleus (pPBcl) and the outer-rostral portion of the external lateral subnucleus (pPBel) projects primarily to the lateral portion of the Ce (CeL). 3) The region of the pPB mainly including the dorsolateral subnucleus (pPBdl), the remaining pPBel, and the external medial (pPBem) subnuclei projects primarily to the lateral capsular portion of the Ce (CeLC) and bilaterally to its rostral portion. Dense projections were also found in the regions which extend the CeLC rostrally and in the SId/GPv, SStr, and FStr rostral areas.The possible role of each of the three parabrachio-amygdaloid pathways described is discussed. It was suggested that the pPB-CeM pathway is mainly implicated in gustatory processes; the pPB-CeL pathway is mainly implicated in visceral and chemosensitive processes; and the pPB-CeLC pathway is mainly implicated in respiratory, cardiovascular, and nociceptive processes.

It has recently been demonstrated that the nucleus centralis of the amygdala contains numerous neurons specifically driven by noxious stimuli. The aim of the present study was to investigate the effect of i.v. morphine on responses of neurons located in the nucleus centralis of the amygdala to noxious mechanical or thermal stimuli. It was observed, in halothane-anesthetized rats, that i.v. morphine caused a marked depression of responses induced by noxious thermal (waterbath, 50 degrees C) and mechanical (pinch) stimuli and caused a moderate depression of spontaneous activity in a dose-related (1, 3, 9 mg/kg) and naloxone reversible fashion. The ED50 value was 1.2 and 9 mg/kg for i.v. morphine for the evoked activity and spontaneous activity, respectively. The strong depressive effect of morphine on evoked activity probably reflects a direct action of this drug at both spinal and parabrachial levels. These results could account, at least in part, for the effect of morphine on the emotional-affective aspects of pain.

1. Neurons (n = 177) were recorded with extracellular micropipettes in and around the nucleus centralis of the amygdala (Ce), in anesthetized rats. The spontaneous activity of these neurons was variable (0.25 less than 3 less than 35 Hz, n = 175; 10th percentile less than median less than 90th percentile). A majority (80%) of these neurons were excited or inhibited exclusively or preferentially by noxious stimuli. These units were separated into two groups: 1) a group of neurons excited by noxious stimuli (46% of the whole population) and 2) a group of neurons inhibited by noxious stimuli (34% of the whole population). 2. The receptive fields of both groups of neurons were very large: in about one-half the cases the neurons responded similarly from all parts of the body, and in the other cases the responses were greater when the stimuli were applied to a restricted part of the body. 3. Seventy-seven percent of the excited neurons had responses of relatively high magnitudes. In this group, most cells (75%) were exclusively driven by noxious stimuli; the others (25%) were preferentially activated by noxious stimuli. These neurons responded to mechanical (pinch or squeeze) and/or thermal (water bath or water jet greater than 44 degrees C) noxious stimuli with a marked and sustained activation. 4. Sixty percent of the inhibited neurons had a marked decrease of activity in response to noxious stimuli. In this group, most of them (81%) were exclusively inhibited by noxious stimuli, whereas the remainder (19%) were preferentially inhibited by noxious stimuli. These neurons responded to mechanical (pinch or squeeze) and/or thermal (water bath or waterjet greater than 44 degrees C) noxious stimuli with a suppression or a marked and sustained decrease in activity. 5. All of the nociceptive neurons responded to intense transcutaneous electrical stimulation with one or several components of activation or inhibition. According to their latencies, three types of components were distinguished: early, intermediate, and late components. We estimate that the early and the intermediate components would be triggered by the activity of peripheral fibers in the 6- to 20-m/s range and therefore could be in the A delta fibers range, whereas the late component would be triggered by fibers in the 0.5- to 1-m/s range and therefore could be in the C fibers range. 6. The neurons excited or inhibited by noxious stimuli were not homogeneously distributed in and around the Ce.(ABSTRACT TRUNCATED AT 400 WORDS)

Small iontophoretic applications of Phaseolus vulgaris leucoagglutinin (PHA-L) were used to study the ascending efferent projections in the rat from the external parabrachial (PBe) area (i.e. external lateral (PBel) and external medial (PBem) subnuclei). It was found that fibers of the caudal two third of PBe project mainly to nucleus centralis of the amygdala (Ce) with a precise pattern: the PBel subnuclei mainly project to the caudomedial subdivision of the Ce and the PBem subnuclei mainly project to the rostrolateral subdivision of the Ce. Another dense and common projection of both subnuclei was found in the ventral pallidal area adjacent to the Ce. This study delineates and extends the terminal area of the spino(trigemino)-ponto-amygdaloid nociceptive pathway demonstrated by our previous studies.

The release of [3H]γ-aminobutyric acid ([3H]GABA) continuously formed from [3H]glutamine has been measured with a push—pull cannula implanted in the substantia nigra of the rat anesthetized with ketamine. Consistent with the high density of GABA terminals coming from both the striato-pallido-nigral afferents, and from GABA nigrofugal neurons, our results showed that a large amount of [3H]GABA was spontaneously released in the reticulata, about 4 times higher than in the compacta. In the absence of calcium the spontaneous [3H]GABA release was reduced (−30%), as well as the K+-induced release of [3H]GABA (−66%). Bicuculline (10−4 M) did not affect the K+-evoked release of [3H]GABA, suggesting that autoreceptors on GABA afferent fibers, are distinct from the GABAA subtype. Partial lesions of striato- and pallido-nigral GABA neurons with kainic acid (1.2 μg) decrease by 40% the glutamic acid decarboxylase (GAD) activity in the ipsilateral SN without decreasing the spontaneous release of [3H]GABA; even following extensive lesions with kainic acid (2.5 μg), GAD activity (−72%) and spontaneous [3H]GABA release (−83%) were not completely abolished. These results suggest that a non-negligible contribution of GABA nigral neurons accounts for the spontaneous GABA release measured in the substantia nigra. This is further supported by the decrease (−20%), and the increase (+40%) of [3H]GABA release produced by the local application of glycine (10−6 M), and bicuculline (10−4 M), which respectively, inhibits and activates the nigral neuron activity. The contribution of nigral GABA neurons to the amount of [3H]GABA release from the substantia nigra, is likely to their high spontaneous firing rate.

The effect of administering low doses (0.5-1.5 micrograms) of the mu-opioid receptor agonist fentanyl into the right brachial plexus sheath of the rat was examined using the vocalization threshold to paw pressure test. Both forepaws were tested in each rat. Fentanyl injected into the right brachial plexus sheath at 0.5-1.5 micrograms/kg produced a localized, dose-dependent, potent and long lasting antinociceptive effect, as gauged on the right forepaw. At the lower dose used (0.5 microgram/kg of fentanyl), the antinociceptive effect was restricted to the right forepaw and lasted for more than 2 h. Increasing doses of fentanyl (1 and 1.5 micrograms/kg) induced potent effects, lasting up to 5-6 h or even longer. In complete contrast, fentanyl administered i.v. at the dose of 1 microgram/kg had a very transient effect, only lasting up to 25 min. The results of injection of low doses of the opioid antagonist naloxone when administered either i.v. or locally into the paw, on the effect of fentanyl suggest the involvement of a peripheral site of action of the opioid. The present findings suggest that, as already observed in patients in clinical situations, low doses of opiates delivered using this administration route may provide prolonged regional analgesia, with the potential of avoiding centrally mediated side effects.

Incidence rates of cutaneous malignant melanoma (CMM) have increased worldwide. Long-term studies examining rates and anatomic site-specific incidence on a population-based level are infrequent.We sought to examine the historical changes in the incidence and anatomic site presentation of CMM during a 50-year period in Manitoba, Canada.Using population-based data, all first diagnoses of CMM reported between 1956 and 2005 were identified. Age-specific rates, age-standardized incidence rates, and anatomic sites were recorded.Incidence rates of CMM slowed for each sex beginning in 1981 for female patients and 1992 for male patients. Annual percent change revealed decreasing rates among male patients younger than 40 years (1992-2005: –5.3% [P = .03]) and female patients younger than 40 years (1987-2005: –1.8% [P = .15]). Similarly, middle-aged individuals (age 40-59 years) also had diminished annual percent change (men 1992-2005: 0.6% [P = .65]; women 1983-2005: –0.3% [P = .68]). The annual percent change for older men and women (60-79 and ≥80 years) continued to increase. Anatomic site-specific analyses revealed that the trunk was the most frequent site of CMM for young male patients (<60 years) whereas the lower extremities were the most common among young female patients (<60 years). Incidence rates for each site, however, are slowing. Among those aged 60 years and older, the rates for each anatomic site increased.Determining changes in tumor thickness would have been useful in determining whether the nature of tumors have changed over time; however, this is not recorded in our registry.The rates of CMM are slowing; however, this change is confined to younger individuals. Anatomic site-specific CMMs are changing; rates among older individuals continue to increase for both sexes.

Several studies have considered the establishment of vascularization in intracerebral solid transplants of neural tissue. The widely supported interpretation of the results is that the vascular network of the solid grafts is already present before implantation into the host brain. The situation is different when dissociated fetal tissue is transplanted as a cell suspension because in these conditions the fetal vascular network is disrupted. The present study has, therefore, been undertaken to follow the angiogenesis in a transplant of dissociated fetal cells implanted into the excitotoxically neuron-depleted thalamus. The vascular network is compared to that observed in the intact and in the lesioned thalamus both in terms of morphology of the capillaries and of the function of the blood-brain barrier (BBB). In the transplant, capillaries, stained by Indian ink, are very few in number and have very fine calibers during the first 20 days after grafting. Some structures can be identified as immature blood vessels at the electron microscopic level. The blood vessels are progressively more numerous in the graft and they demonstrate mature ultrastructural features 2 months after grafting. Last, there is no leakage of the BBB for peroxidase. The vascularization seems to follow a pattern of maturation comparable to that described during development in the literature. In contrast, in the lesioned area, there is a reactive angiogenesis: 10 days after the excitotoxic injection (shortest time studied), there are many wide caliber vessels with expanded perivascular spaces engorged with mesodermal cells. A microvascularization also develops transiently during the first two months. Capillaries are abnormal from the functional point of view, since there is a leakage of the BBB to macromolecules. The use of an experimental model in which transplant had to grow in a lesioned area permits to determine two types of vascularization: an apparently normal developmental timetable, normal morphological and functional characteristics, in the transplant; a reactive angiogenesis, in the lesioned area.

In a previous study, it was shown that, one month after kainic acid (KA) injection into the thalamus, afferents deprived of postsynaptic target neurons exhibit structural alteration, including the loss of synaptic vesicles. The present study was undertaken to determine whether these long-term morphological changes were associated with changes in biochemical markers of monoaminergic pathways. In situ injection of KA was performed into the right ventrobasal complex of the rat thalamus (VB). Protein content and total amount of norepinephrine, dopamine, 5-hydroxytryptamine and 5-hydroxyindolacetic acid were analyzed in the lesioned area one, two, three and four months after injection using high-performance liquid chromatography and electrochemical detection. The results were compared to those obtained in an equal volume of tissue dissected out from the opposite (intact) VB. Protein content per unit volume decreased progressively to 50% of control in the neuron-depleted area. In contrast, whatever the amine considered, its total amount remained unaltered up to 4 months after the excitotoxic lesion. 5-hydroxyindolacetic acid was also unchanged 4 months after lesion. This study suggests that (i) the quantity of monoamines in afferents to the rat VB does not depend upon the presence of post-synaptic target neurons, (ii) a non-vesicular storage compartment may compensate the loss of synaptic vesicles in afferent fibers to the lesioned area.

In the anaesthetized rat the intravenous injection of 5-HT antagonists, cinanserin and methysergide, induces in two-thirds of neurones studied, an increase in the responsiveness of dorsal horn convergent neurones to C-fibre stimulation. These results are in favour of the existence of serotonergic tonic descending inhibitory effects on the spinal transmission of noxious messages.

The purpose of the present study was to investigate whether the level of norepinephrine and its rate of disappearance after decarboxylase inhibition were modified in the spinal cord of a chronic pain model: the arthritic rat. Chromatographic studies allowing the simultaneous determination of norepinephrine and uric acid by means of HPLC with electrochemical detection are described. The norepinephrine and uric acid levels in the spinal cord were higher in arthritic rats than in normal rats. In addition the rate of disappearance of the amine was increased in the dorsal part of the cord in arthritic rats. These results agree with previous reports suggesting an activating effect of nociceptive stimuli on descending noradrenergic systems. They also indicate that studies on purine metabolism in the CNS in inflammatory and/or pain processes will be of interest.