Hippocampal input to preoptic thermosensitive neurons in the rat

The effects of electrical stimulation of ventral subiculum (VSB) of the hippocampus of the thermosensitive neurons in the preoptic area were studied in urethane-anesthetized rats. VSB stimulation affected thermosensitive neurons more frequently (92.1%, 58 of 63) than thermally insensitive neurons (71.4%, 55 of 77). The majority of the VSB-responsive thermosensitive neurons (33 of 44 warm-units and 11 of 14 cold-units) were initially inhibited following stimulation. The result provides further support for the involvement of hippocampus in the central control of thermoregulation.     

Responsiveness of solitario-parabrachial relay neurons to taste and mechanical stimulation applied to the oral cavity in rats

Summary A total of 311 units, responsive to natural stimulation of the oral cavity, were isolated from the rostral part of the solitary tract nucleus (NTS) of rats. Of these, 169 taste neurons, activated by taste stimulation, and 142 mechanoreceptive units, exclusively sensitive to mechanical stimulation of the oral cavity, were found. Most taste units (62.3%) were also excited by mechanical stimulation. Forty-three (34.1%) of the 126 taste units examined were identified as solitario-parabrachial relay (SP) neurons by antidromic stimulation from the ipsilateral dorsal pons, while only eleven (12.6%) of the 87 mechanoreceptive units were SP neurons. Taste SP neurons could be divided into two subgroups according to their antidromic latency; the fast SP units with an antidromic latency shorter than 9 ms and slow SP units with a longer antidromic latency. These two subgroups were not differentiated in any physiological properties except that the fast SP neurons were frequently excited by sucrose. Taste neurons were classified according to the best stimulus of the four basic taste solutions to produce the largest number of discharges in each neuron. All types of taste neurons were found among the SP and non-SP neurons, but only a small number of quinine-best neurons (n = 2) were found in the SP neuron group compared to the number of quinine-best neurons in the non-SP neuron group (n = 10). A histological examination of the recording sites revealed that taste relay neurons were found at the central or dorsal part of the nucleus but mechanoreceptive relay neurons were found at the peripheral part, although relay and non-relay neurons of either class were intermingled in the nucleus.Supported by a Grant from the Ministry of Education, Science and Culture of Japan (No. 58106008)     

Hypothalamic warm-sensitive neurons possess a tetrodotoxin-sensitive sodium channel with a high Q10

We have investigated the ionic current responses to temperature of dissociated cells from the preoptic and anterior hypothalamus (PO/AH) of rat, using the 'whole-cell' configuration. The majority of the recorded neurons showed a linear increase in a non-inactivating inward current during warming (30-40 degrees C), and the Q10 was about 2. However, about 24% of PO/AH neurons were markedly sensitive to warming and the increase in non-inactivating inward current to a rise in temperature in the hyperthermic range (35-40 degrees C) had a high Q10 (4.3-7.0). This increase in current in the hyperthermic range was reversibly blocked by tetrodotoxin (TTX). The inward current in neurons with a Q10 of 2 was not affected by TTX. The results show that some neurons in the PO/AH possess a non-inactivating sodium channel that is highly temperature-sensitive in the hyperthermic range. These neurons are presumably the 'primary' warm-sensitive neurons.     

Responses of preoptic thermosensitive neurons to poikilothermia-inducing peptides — bombesin and neurotensin

Microelectrophoretic application of bombesin (BS) and neurotensin (NT) to thermosensitive and non-thermosensitive neurons in the preoptic and anterior hypothalamus (PO/AH) of anesthetized rats depressed the activity of 78 (54.2 %) of 144 neurons and that of 66 (51.6 %) of 128 neurons, respectively. The depressing effects of both peptides which were uniformly observed across warm-sensitive and cold-sensitive neurons in the PO/AH are consistent with their poikilothermic actions.     

Nucleus accumbens and preoptic area stimulation: tuberoinfundibular single unit responses,modulation of electrical activity and gonadotrophin secretion

Summary Single unit activity was recorded from tuberoinfundibular neurones in urethane-anaesthetized pro-oestrous rats. Responses following single shock (0.2 Hz) stimulation of the preoptic area/ anterior hypothalamus (PO/AH) or nucleus accumbens (ACB) were recorded and computed for 137 single units. More cells were shown, by antidromic activation, to project to ACB (4 of 77) than to PO/ AH (1 of 60). Fewer units were responsive to stimulation of ACB (p < 0.02), those that were responsive showing a longer latency to the onset of response compared to that following PO/AH stimulation. Significantly (p < 0.02) more slowly firing cells were found in the group of animals tested with stimulation of PO/AH. Delivery of high frequency stimulation to PO/AH resulted in a 2 1/2 fold increase in plasma luteinizing hormone (LH) levels (p < 0.05), whereas ACB-stimulated and control animals showed no increases in plasma LH concentrations. During delivery of the high frequency stimulation only one cell in the ACB group of animals was orthodromically affected (out of five tested). However, in the PO/AH group, five of the seven cells tested showed either excitation or inhibition during stimulation. The possibility that the functional connections demonstrated between ACB and the endocrine hypothalamus are involved in the expression of higher brain functions, recognised during changing endocrine states, is discussed.     

Multiple innervation by substance P-containing fibers in the parabrachial area of the rat

The present study demonstrates that a large number of substance P (SP) fibers in the parabrachial area (PB) of the rat originate from at least three sources. The majority of SP fibers in the lateral surface of the lateral parabrachial area (PBLI) and medical parabrachial area originate from SP neurons located caudal to the PB. Some of the SP fibers in the PBLI originate from SP neurons located rostral to the PB. SP fibers in the ventral part of the lateral parabrachial area originate from SP cells from the pons at the level of the PB.     

Tissue levels and in vivo release of tachykinins and GABA in striatum and substantia nigra of rat brain after unilateral striatal dopamine denervation

Summary Brain tissue levels and in vivo release of substance P (SP) and neurokinin A (NKA) and GABA were measured bilaterally in striatum and substantia nigra of the rat, after a unilateral 6-hydroxydopamine lesion of the nigro-striatal dopamine pathway. Sham injected animals served as controls. The dopamine denervation decreased the tissue levels of SP in striatum (-38%) ipsilateral to the lesion and in substantia nigra both ipsi- (-54%) and contralateral (-38%) to the lesion. NKA was not significantly changed in the striatum, but decreased (like SP) in the substantia nigra both ipsi- (-50%) and contralateral (-40%) to the lesion. GABA tissue levels increased in the denervated striatum (+20%) and remained unchanged in substantia nigra at both sides. The extracellular levels of SP, NKA and GABA were measured with microdialysis in vivo at basal conditions and during stimulation with potassium administered locally via the microdialysis probe. The stimulated release of SP and NKA in the substantia nigra ipsilateral to the lesion was compared to in sham operated animals reduced with 39% and 64%, respectively, while no change in SP or NKA release was detected in the striatum. The basal release of GABA in the striatum was increased with 296% and with 76% during stimulation in the dopamine denervated striatum, while no change in GABA basal or stimulated release was detected in the substantia nigra. We suggest that the increased GABA release in the dopamine denervated striatum may be due to a decreased dopamine mediated inhibition of local GABA neurons. Furthermore, the decreased nigral release of SP and NKA ipsilateral to the lesion is suggested to be caused by an increased GABA inhibition in striatum of SP- and NKA-containing striato-nigral neurons.     

Substance P modulates glutamate-induced currents in acutely isolated rat spinal dorsal horn neurones.

The whole-cell patch-clamp technique was used to examine the effect of substance P (SP) on glutamate-induced currents in freshly dissociated rat spinal dorsal horn neurons (LI-III). In 48% of examined cells SP (10(-10)-10(-6) M) at -70 mV, induced in inward current that desensitized in the continued presence of SP. When applied simultaneously with, or prior to L-glutamate, SP caused a potentiation of L-glutamate-induced current in 65% of the tested cells. Since glutamate activates both N-methyl-D-aspartate (NMDA) and non-NMDA receptors in rat dorsal horn neurons, selective agonists, kainate, quisqualate, alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) and NMDA were used to determine which subtype of excitatory amino acid receptors interacted with SP. We found that the responses to quisqualate, kainate, and AMPA were not significantly affected by SP (less than 20% increase). In contrast, the inward currents induced by NMDA (30-300 microM) appear to be reduced and potentiated after the administration of 2-200 nM of SP. These results suggest that post-synaptic mechanisms of action of tachykinins may contribute to the regulation of the strength of glutamate-mediated excitatory transmission in the rat spinal dorsal horn.     

Temporomandibular joint inflammation potentiates the excitability of trigeminal root ganglion neurons innervating the facial skin in rats

The aim of this study was to test the hypothesis that temporomandibular joint (TMJ) inflammation alters the excitability of trigeminal root ganglion (TRG) neurons innervating the facial skin, by using behavioral, electrophysiological, molecular, and immunohistochemical approaches. Complete Freund's adjuvant (CFA) was injected into the rat TMJ to produce inflammation. The threshold for escape from mechanical stimulation applied to the orofacial area in TMJ-inflamed rats was significantly lower than that in na?ve rats. The TRG neurons innervating the inflamed TMJ were labeled by 2% Fluorogold (FG) injection into the TMJ. The number of FG-labeled substance P (SP)-immunoreactive neurons in the inflamed rats was significantly increased compared with that in the na?ve rats. On the other hand, medium- and large-diameter TRG neurons (>30 microm) innervating the facial skin were labeled by FG injection into the facial skin. In the FG-labeled cutaneous TRG neurons, the occurrence of SP (100 nM) induced membrane depolarization in inflamed rats (medium: 73.3%, large : 85.7%) was larger than that in the na?ve rats (medium: 29.4%, large : 0%). In addition, SP application significantly increased the firing rate evoked by depolarizing pulses in the neurons of inflamed rats compared with those of na?ve rats. Quantitative single-cell RT-PCR analysis showed the increased expression of mRNA for the NK1 receptor in FG-labeled TRG neurons in inflamed rats compared with that in naive rats. The numbers of SP and NK1 receptors/neurofilament 200 positive immunoreactive TRG neurons innervating the facial skin (FG-labeled) in the inflamed rats were significantly increased compared with those seen in na?ve rats. These results suggest that TMJ inflammation can alter the excitability of medium- and large-diameter TRG neurons innervating the facial skin and that an increase in SP/NK1 receptors in their soma may contribute to the mechanism underlying the trigeminal inflammatory allodynia in the TMJ disorder.     

Effects of peripheral chemo- and baro-receptor denervation on responses of preoptic thermosensitive neurons to inspired CO2

The purpose of this study was to see if the responses of thermosensitive neurons in the preoptic (PO) area to inspired CO₂ seen in spontaneously ventilated rats were indirectly driven by reflexive changes in respiration and circulation. In urethanized, paralyzed, and artificially ventilated (AV) rats, the effects of 10% CO₂ inhalation on PO thermosensitive neurons were examined by regression of neuronal activity on PO temperature. The experiments were made in intact rats and in rats whose peripheral chemo- and baro-receptors were denervated (AVD). In both AV and AVD rats, the slope of the regression line decreased significantly (P<0.05) during CO₂ inhalation in half of the warmsensitive neurons studied (64.3% in AV rats, 41.7% in AVD rats). Peripheral chemo- and baro-receptors thus do not appear to be responsible for decreased thermosensitivities of warm-sensitive neurons during CO₂ inhalation. The tendency for activities of warm-sensitive neurons to increase progressively at lowerT _po was seen during CO₂ inhalation in both AV and AVD rat. However, the average differences in mean firing rate between 10% CO₂ and air inhalations were 2–3 imp/s greater at anyT _po in AV rats than in AVD rats. In AVD rats, warm-sensitive neurons were rather inhibited by CO₂ at higherT _po. Excitation of warm-sensitive neurons during CO₂ inhalation in AV rats, which was independent ofT _po, was considered to be caused by the signals from peripheral receptors.     

5-HT1A受体亚型与P物质、Ⅰ型囊泡膜谷氨酸转运体和甘丙肽在大鼠背根神经节神经元中的共表达

为探讨5-HT1A受体亚型参与感觉信息调控的机制,本文利用免疫荧光组织化学双重染色技术观察了该受体亚型与P物质(SP)、I型囊泡膜谷氨酸转运体(VGLUT1)和甘丙肽(Gal)在大鼠背根神经节(DRG)神经元内的共存状况。结果表明:5-HT1A受体亚型阳性神经元占DRG神经元总数的46.2%,阳性神经元以大型及小型神经元为主。在DRG内观察到了5-HT1A/SP、5-HT1A/VGLUT1以及5-HT1A/Gal双标神经元。其中5-HT1A/SP双标神经元占5-HT1A受体亚型阳性神经元的34.6%,占SP阳性神经元的72.0%;5-HT1A/VGLUT1双标神经元占5-HT1A受体亚型阳性神经元的24.1%,占VGLUT1阳性神经元的18.5%;5-HT1A/Gal双标神经元占5-HT1A免疫阳性神经元的17.6%,占Gal免疫阳性神经元的63.8%。5-HT1A/SP和5-HT1A/Gal双标神经元主要为DRG的小型神经元,而5-HT1A/VGLUT1双标神经元主要为大、中型神经元。上述结果提示,5-HT1A受体亚型可能通过调节SP、谷氨酸以及Gal在初级传入终末及外周神经末稍的释放发挥其感觉信息的调节作用。     

P物质对大鼠DRG神经元H+门控电流的调制作用

 目的：观察P物质（SP）对神经元H+门控电流的调制作用及其可能存在的机制。方法：采用全细胞膜片钳技术分别记录pH 4.0、pH 5.0、pH 6.0及共加H+ 与SP情况下,急性分离的大鼠背根神经节（DRG)神经元H+门控电流的形式。通过胞内透析技术分析SP对H+门控电流可能的调控机制。结果：H+门控电流可划分为短暂内流型（T型）、持续内向电流型（S型）、反向电流型（O型）及双相内向电流型（B型）。SP对S型和B型中持续成分的H+门控电流有明显增强作用,这种电流幅值增强达（85.53±22.93）%,约81.8%细胞的这种增强效应不能被选择性的SP受体NK1拮抗剂阻断,而非肽类的SP受体NK1拮抗剂对约75%的DRG神经元可明显阻断这种增强效应;SP对S型和B型中持续成分的H+门控电流也有明显抑制作用,抑制的幅值达（48.46±4.45）%,并且约88.9%细胞的这种抑制效应不能被SP受体NK1拮抗剂所阻断。通过胞内透析技术在细胞内液中加入GDP-β-S后不能消除SP对H+门控电流的调制作用。结论：H+门控电流有T型、S型、O型及B型4种电流类型。SP对H+门控电流有增强和抑制的双向调节作用。SP可能通过G蛋白偶联受体通路及与H+门控离子通道的某一位点直接结合来发挥对H+门控电流的双向调节调制作用。     

Spatial association of prolyl oligopeptidase, inositol 1,4,5-triphosphate type 1 receptor, substance P and its neurokinin-1 receptor in the rat brain: an immunohistochemical colocalization study

Prolyl oligopeptidase (POP) is a serine endopeptidase which hydrolyzes proline-containing peptides shorter than 30 amino acids. It has been suggested that POP is associated with cognitive functions, possibly via the cleavage of neuropeptides such as substance P (SP). Recently, several studies have also linked POP to the inositol 1,4,5-triphosphate (IP(3)) signaling. However, the neuroanatomical interactions between these substances are not known. We used double-labeled immunofluorescence to determine the POP colocalization with SP, SP receptor (neurokinin-1 receptor, NK-1R) and IP(3) type 1 receptor (IP(3)R1) in the rat brain. Furthermore, since striatal and cortical GABAergic neurons are involved in SP neurotransmission, we studied the coexpression of POP, SP and GABA by triple-labeled immunofluorescence. POP was moderately present in IP(3)R1-containing cells in cortex; the colocalization was particularly high in the thalamus, hippocampal CA1 field and cerebellar Purkinje cells. Colocalization of POP with SP and NK1-receptor was infrequent throughout the brain, though some POP and SP coexpression was observed in cerebellar Purkinje cells. We also found that POP partially colocalized with SP-containing GABAergic neurons in striatum and cortex. Our findings support the view that POP is at least spatially associated with the IP(3)-signaling in the thalamus, hippocampus and cerebellar Purkinje cells. This might point to a role for POP in the regulation of long-term potentiation and/or depression. Moreover, the low degree of colocalization of POP, SP and its NK-1R suggests that a transport system is needed either for POP or SP to make hydrolysis possible and that POP may act both intra- and extracellularly.     

Effect of nitric oxide on substance P release from the peripheral endings of primary afferent neurons.

To elucidate the interaction between nitric oxide (NO) and substance P (SP) in neurogenic inflammatory responses, we studied the effect of drugs related to nitric oxide (NO) on the levels of Evans blue and SP released into perfusate from the subcutaneous space in the rat instep. Noxious heat stimulation (47 degrees C for 30 min) caused an increase in SP release in parallel with plasma extravagation. N(omega)-nitro-L-arginine methylester (L-NAME: 100 mg/kg, once daily) given intraperitoneally (i.p.) five times (chronic treatment) significantly suppressed the heat-evoked SP release and Evans blue leakage. FK409 (10(-4) M), which evokes a release of NO, applied locally caused a remarkable increase in the SP release into the perfusate. These results suggest that heat-induced NO generation causes SP release from the peripheral endings of small-diameter primary afferent neurons.     

Physiological characteristics of the solitario-parabrachial relay neurons with tongue afferent inputs in rats

Summary Among 180 units in the solitary tract nucleus (NTS) of rats, 34 solitario-parabrachial relay neurons (SP neurons), were identified by antidromic activation from the parabrachial nucleus. The SP neurons were classified into two groups, fast and slow, according to their antidromic latencies. The responsiveness of the SP in comparison with non-SP neurons was studied by electrical stimulation of three tongue nerves: the lingual (L), chorda tympani (CT) and glossopharyngeal (G) nerves. About half the SP neurons produced a single spike with an orthodromic latency of 2–5 ms, while about one third of them discharged more than two spikes. A few neurons gave rise to a long-lasting discharge consisting of five or six spikes. Some SP neurons were excited by stimulation of the tongue afferents with a low stimulus intensity, but other SP neurons produced spikes at only very high voltages. Fast SP cells were not differentiated from slow SP cells, except that latency of orthodromic responses to CT stimulation was significantly shorter in the former than in the latter (P<0.05, Mann-Whitney U-test). Locations of the SP and non-SP neurons, reconstructed histologically, indicate that they do not distribute evenly throughout the mediolateral extent at the rostral pole of the NTS, but clustered in its medial half.     

Complementary action of substance P and vasoactive intestinal peptide on the rat parotid secretion

Augmentation of the rat parotid salivary secretion to intravenous injections of substance P (SP) occurred when SP was combined with vasoactive intestinal peptide (VIP), or stimulation of the auriculo-temporal nerve in the presence of atropine and the adrenergic blockers, dihydroergotamine and propranolol. The largest increase was obtained when SP (0.5 micrograms kg-1) was used together with subthreshold doses of VIP (84% at 0.05 micrograms kg-1 and 105% at 0.5 micrograms kg-1) and low frequency stimulation (92% at 2 Hz and 97% at 5 Hz), which did not produce any salivary secretion by itself. There was no facilitated secretion when VIP and nerve stimulation were combined. Amylase output was much larger (250-500%) when SP was combined with nerve stimulation (0.5-5 Hz) or VIP (0.005-5 micrograms kg-1) than when SP was used alone. Similar results were obtained in rats where the auriculo-temporal nerve was stimulated during the early phase (24-90 h) of Wallerian degeneration, when the nerve-induced responses were seemingly completely blocked. Our results are consistent with the hypothesis that both VIP and SP contribute to the atropine-resistant parotid secretion, and that they have a complementary role in the rat parotid exocrine function.     

Neurochemical phenotype of vagal afferent neurons activated to express C-FOS in response to luminal stimulation in the rat

The vagus nerve conveys meal-induced primary afferent responses to the brainstem. Electrophysiological studies indicate that luminal stimuli such as osmolarity and the digestion products of carbohydrates elicit powerful vagal nodose neuronal responses by activating serotonin 3 (5-hydroxytryptamine-3, 5-HT3) receptors on intestinal mucosal afferent fibers. To characterize the neurochemical phenotype of neurotransmitters in vagal nodose neurons that are activated by luminal stimulation, we examined c-fos protein (c-Fos) expression in response to luminal stimulation in conscious rats. A double-labeling technique using antisera to glutamate (Glu), substance P (SP), calcitonin gene-related peptide (CGRP), and somatostatin (SS) was used to determine the neurochemical profile of c-Fos-positive neurons. c-Fos immunoreactivity was insignificant in vehicle-treated rats. Luminal perfusions of NaCl (500 mOsm), tap water (5 mOsm), maltose (300 mmol/l), and 5-HT (10(-5) mol/l) each elicited a significant increase in the number of cells expressing c-Fos. Chronic vagotomy eliminated an increase in nodose neuronal c-Fos expression, and the 5-HT3 receptor antagonist granisetron significantly reduced it. Glu-, SP-, and CGRP-containing neurons represented 28%, 53%, and 19%, respectively, of the total population of nodose neurons. Few neurons contained SS. Double-labeling studies revealed that of the c-Fos-positive neurons responsive to hypertonic NaCl, 52%, 41%, and 3% exhibited immunoreactivity for Glu, SP, and CGRP, respectively. Of those responsive to tap water, 47%, 50%, and 4% exhibited immunoreactivity for Glu-, SP- and CGRP, respectively. In addition, 44%, 38%, and 8% of 5-HT-stimulated and 30%, 32%, and 5% of maltose-stimulated c-Fos-positive neurons exhibited, respectively, Glu, SP, and CGRP immunoreactivity. The few neurons that contained SS did not express c-Fos. CONCLUSIONS: Vagal primary afferent neurons that respond to 5-HT-dependent luminal stimuli, such as hyperosmolarity and maltose, contain mainly Glu and SP. These neurons appear to play an important role in the mediation of the vago-vagal reflex elicited by luminal stimuli.     

Single-cell RT-PCR, in situ hybridization histochemical, and immunohistochemical studies of substance P and enkephalin co-occurrence in striatal projection neurons in rats

Single-cell RT-PCR studies in 3-4-week-old rats have raised the possibility that as many as 20% of striatal projection neurons may be a unique type that contains both substance P (SP) and enkephalin (ENK). We used single-cell RT-PCR, retrograde labeling, in situ hybridization histochemistry, and immunolabeling to characterize the abundance of this cell type, its projection target(s), and any developmental changes in its frequency. We found by RT-PCR that 11% of neurons containing either SP or ENK contained both in 4-week-old rats, while in 4-month-old rats SP/ENK colocalization was only 3%. SP-only neurons tended to co-contain dynorphin and ENK-only neurons neurotensin, while SP/ENK neurons tended to contain dynorphin. Single-cell RT-PCR showed SP/ENK co-occurrence in 4-week-old rats to be no more common among striatal neurons retrogradely labeled from the substantia nigra than among those retrogradely labeled from globus pallidus. Double-label in situ hybridization showed SP/ENK perikarya to be scattered throughout striatum, making up 8% of neurons containing either SP or ENK at 4 weeks, but only 4% at 4 months. Immunolabeling showed that presumptive striatal terminals in globus pallidus externus, globus pallidus internus and substantia nigra pars reticulata that colocalized SP and ENK were scarce. Terminals colocalizing SP and ENK were, however, abundant in the substantia nigra pars compacta. Thus, SP-only and ENK-only neurons make up the vast majority of striatal projection neurons in rats, the frequency of SP/ENK colocalizing striatal neurons is low in adult rats (3-4%), and SP/ENK colocalizing neurons primarily project to SNc but do not appear to be confined to striosomes.     

Oxygen sensitivity of NMDA receptors: relationship to NR2 subunit composition and hypoxia tolerance of neonatal neurons

Neonatal rats survive and avoid brain injury during periods of anoxia 25 times longer than adults. We hypothesized that oxygen activates and hypoxia suppresses NMDA receptor (NMDAR) responses in neonatal rat neurons, explaining the innate hypoxia tolerance of these cells. In CA1 neurons isolated from neonatal rat hippocampus (mean postnatal age [P] 5.8 days), hypoxia (PO(2) 10 mm Hg) reduced NMDA receptor-channel open-time percentage and NMDA-induced increase in [Ca(2+)](i) (NMDA DeltaCa(2+)) by 38 and 68% (P<0.01), respectively. In P20 neurons the reductions were not significant. In P3-10 CA1 neurons within intact hippocampal slices, hypoxia reduced NMDA DeltaCa(2+) by 52% (P=0.002) and decreased NMDA-induced death by 45% (P=0.004). Phalloidin, a microtubule stabilizer, prevented hypoxia-induced inhibition of NMDA DeltaCa(2+) in P3-10 neurons. To test whether NMDARs prevalent in neonates (NR1 plus NR2B or NR2D subunits) are inhibited by hypoxia compared with those in mature neurons (NR2A and NR2C), we expressed these receptors in Xenopus oocytes. Compared with responses in 21% O(2), hypoxia (PO(2) 17 mm Hg) reduced currents from neonatal type NR1/NR2D receptors by 25%, increased currents from NR1/NR2C by 18%, and had no effect on NR1/NR2A or NR1/NR2B. Modulation of NMDARs by hypoxia may play an important role in the hypoxia tolerance of the mammalian neonate. In addition, oxygen sensing by NMDARs could play a significant role in postnatal brain development.     

Stress induces substance P in vagal sensory neurons innervating the mouse airways

BACKGROUND: Tachykinins-like substance P (SP) have been shown to play an important role in initiating and perpetuating airway inflammation. Furthermore, they are supposed to be released into tissues in response to stress. OBJECTIVE: The aim of this study was to investigate the effects of stress alone or in combination with allergic airway inflammation on SP expression in sensory neurons innervating the mouse airways. METHODS: Balb/c mice were systemically sensitized to ovalbumin (OVA), followed by allergen aerosol exposure, and compared with non-sensitized controls. Additionally, OVA-sensitized and -challenged and non-sensitized mice were exposed to sound stress. SP expression in airway-specific and overall vagal sensory neurons of the jugular and nodose ganglion complex was analysed using retrograde neuronal tracing in combination with immunohistochemistry. Preprotachykinin A (PPT-A) mRNA, the precursor for SP, was quantified in lung tissue by real-time PCR. Bronchoalveolar lavage (BAL) fluid was obtained, and cell numbers and differentiation were determined. RESULTS: Stress and/or allergic airway inflammation significantly increased SP expression in retrograde-labelled vagal sensory neurons from the mouse lower airways compared with controls [stress: 15.7+/-0.8% (% of retrograde-labelled neurons, mean+/-SEM); allergen: 17.9+/-0.4%; allergen/stress: 13.1+/-0.7% vs. controls: 6.3+/-0.3%]. Similarly, SP expression increased in overall vagal sensory neurons identified by the neuronal marker protein gene product (PGP) 9.5 [stress: 9.3+/-0.6% (% of PGP 9.5-positive neurons, means+/-SEM); allergen: 12.5+/-0.4%; allergen/stress: 10.2+/-0.4% vs. controls: 5.1+/-0.3%]. Furthermore, stress significantly increased PPT-A mRNA expression in lung tissue from OVA-sensitized and -challenged animals, and immune cells were identified as an additional source of SP in the lung by immunohistochemistry. Associated with enhanced neuronal SP expression, a significantly higher number of leucocytes were found in the BAL following allergen exposure. Further, stress significantly increased allergen-induced airway inflammation identified by increased leucocyte numbers in BAL fluids. CONCLUSION: The central event of sound stress leads to the stimulation of SP expression in airway-specific neurons. However, in sensitized stressed mice an additional local source of SP (probably inflammatory cells) might enhance allergic airway inflammation.