The use of throat packs in ear, nose and throat, oral and dental surgery: a systematic review.

BACKGROUND: Throat packs are placed around the airway in patients undergoing upper airway surgical procedures under general anaesthetic to prevent aspiration or ingestion of blood, and consequent chest infections or postoperative nausea and vomiting (PONV). There is no definitive evidence for this, and each time a pack is placed, it risks being retained and obstructing the upper airway. This study aimed to determine whether throat packs are of benefit to patients undergoing upper airway surgical procedures. METHODS: Medline, Embase and Central were searched from conception to 15th January 2018 using individualised search strategies. A systematic search of multiple databases was undertaken using custom strategies to identify all relevant randomised controlled trials. Screening, risk of bias assessment and data extraction were undertaken independently by two authors. Primary outcomes included throat pain and PONV. Secondary outcomes included any adverse event documented. RESULTS: Thirteen papers were eligible for inclusion. No studies reported any instances of retained throat packs. Ten studies assessed the effect of throat packs on post-operative throat pain, with four papers showing a significantly higher incidence of pain when throat packs were used. One study showed throat pain to be slightly, but significantly, worse 24 hours post-surgery when a pack was not used. No paper showed throat packs to be of benefit in preventing PONV. CONCLUSION: The study was limited by methodological flaws of included trials, overall relatively low numbers of patients and difficulty in contacting authors to clarify information and obtain raw data. However, this systematic review found no evidence to support the use of throat packs. This supports the proposal that there is no indication for the routine use of throat packs in ENT, maxillofacial and dental procedures.

Iron fertilization to enhance tolerance mechanisms to copper toxicity of ryegrass plants used as cover crop in vineyards.

Ryegrass (Lolium perenne L.) is a plant species that can express mechanisms of tolerance to copper (Cu) toxicity. Therefore, the agronomical approach of intercropping system with ryegrass may represent a promising tool to limit the onset of Cu toxicity symptoms in the other intercropped plants species, particularly when an inadequate nutrient availability like iron (Fe) shortage is also concurrently present. This study aimed at assessing the mechanisms involved in the mitigation of Cu phytotoxicity and the stress effects on plant growth, root morphology and nutrition of ryegrass fertilized with two different Fe sources. To this purpose, seedlings of ryegrass were hydroponically grown for 14 days in controlled conditions with 4 different levels of Cu (0.2, 5.0, 25 and 50 µM) and with either 100 µM Fe-EDDHA or Fe-EDTA. Results show that high levels of Cu availability enhanced the root content of organic anions as well as the root exudation. Different Fe fertilizations at the condition of 50 µM Cu induced changes in root phenolic compounds, citrate and fumarate contents and the exudation pattern of phenolic compounds. Differences in plant growth were not observed between the two Fe sources, although Cu concentration in plant tissue fed with Fe-EDTA was lower in the condition of 50 µM Cu. The enhanced root exudation of Cu-complexing organic compounds (including phenolics) in ryegrass plants when exposed to excessive Cu availability could be at the basis of the ameliorated edaphic rhizosphere conditions (lower Cu availability). For this reason, from the agronomical point of view ryegrass plants used in intercropping systems with crops like vine plants could represent a promising strategy to control Cu toxicity in vineyard soils. Further studies under the field conditions must be taken to support present findings.

Physiological responses of soybean (Glycine max (L.) Merrill) cultivars to copper excess.

Successive applications of copper fungicides on vines have resulted in increased copper content in vineyard soils over the years. This high copper content has affected the growth of young vines in eradicated vineyards. Thus, the cultivation of annual species for a few years is an alternative to copper phytostabilization, because it would be a good way to decrease copper availability to plants. The aim of this study was to evaluate the physiological responses of different soybean cultivars to copper concentration increase. Four different soybean cultivars were grown under three copper concentrations: 0.5, 20 and 40 µM in nutrient solution. The main outcomes of this study were: i) Cultivar M 6410 IPRO recorded the highest photosynthetic rate when plants were exposed to 40 µM of copper in the nutrient solution; ii) plants in cultivar M 6410 IPRO accumulated large copper concentrations in their roots although did not decrease the root dry mass, possibly due to the higher superoxide dismutase activity; iii) cultivar DM 5958 RSF IPRO recorded drastically reduced photosynthetic rate and dry mass production due to copper excess. We conclude that each cultivar responded differently to the excess of copper, but none of them showed tolerance to it.

Measuring dynamic Eustachian tube function using tympanometry in a pressure chamber: the effect of nasal betahistine application.

OBJECTIVE: To assess the effect of topical betahistine on Eustachian tube function in subjectively abnormal subjects in a hyperbaric chamber. METHOD: Active and passive Eustachian tube function was examined using tympanometry in a pressure chamber. RESULTS: Active Eustachian tube function was tested against the negative middle ear pressure induced by increasing the chamber pressure to +3 kPa. One voluntary swallow decreased middle-ear pressure by a mean of 1.36 kPa. Passive Eustachian tube function was tested by measuring spontaneous Eustachian tube openings as the chamber pressure dropped from +10 kPa to ambient. Four distinct patterns of Eustachian tube behaviour were seen, three of which indicated Eustachian tube dysfunction. Betahistine had no positive effect on Eustachian tube opening, although previous animal studies had suggested a beneficial effect. CONCLUSION: Topical betahistine had no effect on Eustachian tube function. Combining a hyperbaric chamber with tympanometry proved ideal for evaluating Eustachian tube function.

From proliferation to target innervation: signaling molecules that direct sympathetic nervous system development.

The sympathetic division of the autonomic nervous system includes a variety of cells including neurons, endocrine cells and glial cells. A recent study (Furlan et al. 2017) has revised thinking about the developmental origin of these cells. It now appears that sympathetic neurons and chromaffin cells of the adrenal medulla do not have an immediate common ancestor in the form a "sympathoadrenal cell", as has been long believed. Instead, chromaffin cells arise from Schwann cell precursors. This review integrates the new findings with the expanding body of knowledge on the signalling pathways and transcription factors that regulate the origin of cells of the sympathetic division of the autonomic nervous system.

Effect of phosphorus on arsenic uptake and metabolism in rice cultivars differing in phosphorus use efficiency and response.

A hydroponic experiment was carried out to investigate the effect of phosphorus (P) nutrition on arsenic (As) uptake and translocation within the seedlings of rice cultivars. The experiment occurred in three stages: I 5 days of acclimatization (nutritive solution); II 10 days under P (0.0 and 0.09 mM) and As (0.0 and 100 mM) treatments; III 5 days under recovery. The As exposure had significant effect reducing dry weights of shoots or roots, resulted in elevated concentrations of As in shoot tissues. BR-IRGA 409 showed the highest susceptibility to As in biomass production and root system parameters regardless the P level. This cultivar showed contrasting responses of As translocation to shoot tissue dependent on P levels, with the highest As concentration under low P and lowest under normal P levels. P nutrition was most striking on plants recovery for all cultivars under As exposure. Clearer separation of cultivars for phosphorus use efficiency (PUE) occurred at lower shoot P contents, that was, at higher levels of P deficiency stress. IRGA 424 showed higher PUE as compared to the others cultivars. Our results go some way to understanding the role of P nutrition in controlling the effects of As in rice shoots.

Effect of phosphorus on arsenic uptake and metabolism in rice cultivars differing in phosphorus use efficiency and response

ABSTRACT A hydroponic experiment was carried out to investigate the effect of phosphorus (P) nutrition on arsenic (As) uptake and translocation within the seedlings of rice cultivars. The experiment occurred in three stages: I 5 days of acclimatization (nutritive solution); II 10 days under P (0.0 and 0.09 mM) and As (0.0 and 100 mM) treatments; III 5 days under recovery. The As exposure had significant effect reducing dry weights of shoots or roots, resulted in elevated concentrations of As in shoot tissues. BR-IRGA 409 showed the highest susceptibility to As in biomass production and root system parameters regardless the P level. This cultivar showed contrasting responses of As translocation to shoot tissue dependent on P levels, with the highest As concentration under low P and lowest under normal P levels. P nutrition was most striking on plants recovery for all cultivars under As exposure. Clearer separation of cultivars for phosphorus use efficiency (PUE) occurred at lower shoot P contents, that was, at higher levels of P deficiency stress. IRGA 424 showed higher PUE as compared to the others cultivars. Our results go some way to understanding the role of P nutrition in controlling the effects of As in rice shoots.

Increased satisfaction after total knee replacement using sensor-guided technology.

The aim of this prospective multicentre study was to report the patient satisfaction after total knee replacement (TKR), undertaken with the aid of intra-operative sensors, and to compare these results with previous studies. A total of 135 patients undergoing TKR were included in the study. The soft-tissue balance of each TKR was quantified intra-operatively by the sensor, and 18 (13%) were found to be unbalanced. A total of 113 patients (96.7%) in the balanced group and 15 (82.1%) in the unbalanced group were satisfied or very satisfied one year post-operatively (p = 0.043). A review of the literature identified no previous study with a mean level of satisfaction that was greater than the reported level of satisfaction of the balanced TKR group in this study. Ensuring soft-tissue balance by using intra-operative sensors during TKR may improve satisfaction.

Genetic fate-mapping of tyrosine hydroxylase-expressing cells in the enteric nervous system.

BACKGROUND: During development of the enteric nervous system, a subpopulation of enteric neuron precursors transiently expresses catecholaminergic properties. The progeny of these transiently catecholaminergic (TC) cells have not been fully characterized. METHODS: We combined in vivo Cre-lox-based genetic fate-mapping with phenotypic analysis to fate-map enteric neuron subtypes arising from tyrosine hydroxylase (TH)-expressing cells. KEY RESULTS: Less than 3% of the total (Hu(+) ) neurons in the myenteric plexus of the small intestine of adult mice are generated from transiently TH-expressing cells. Around 50% of the neurons generated from transiently TH-expressing cells are calbindin neurons, but their progeny also include calretinin, neurofilament-M, and serotonin neurons. However, only 30% of the serotonin neurons and small subpopulations (<10%) of the calbindin, calretinin, and neurofilament-M neurons are generated from TH-expressing cells; only 0.2% of nitric oxide synthase neurons arise from TH-expressing cells. CONCLUSIONS & INFERENCES: Transiently, catecholaminergic cells give rise to subpopulations of multiple enteric neuron subtypes, but the majority of each of the neuron subtypes arises from non-TC cells.

Subcutaneous concentrations following topical iontophoretic delivery of diclofenac.

A self-contained Wearable Electronic Disposable Drug Delivery (WEDD(®)) patch was used to demonstrate that diclofenac levels delivered by iontophoresis are greater than estimated minimal effective concentrations in local subcutaneous tissue and are also greater than either passive transdermal or intravenous delivery using hairless rats. In vitro iontophoretic delivery was evaluated to optimize donor cell formulation using Franz diffusion cells and 1000 NMWL Millipore ultrafiltration membrane. In vivo animal studies were done using patches powered with a 4-volt system, consisting of a 1-volt Zn anode and Ag/AgCl cathode with built in 3-volt lithium battery. Blood and microdialysis samples were collected at different time points after patch application. Current levels increased to 1.0 mA at 30 min, then fell to a steady state of ~ 0.4 mA. Both WEDD(®) and passive patches produced measurable levels of diclofenac in the subcutaneous tissue below the application site (C(max) ± SE = 113.3 ± 61.7 ng/mL and 36.3 ± 15.9 ng/mL, respectively). The dose delivered in six hours was calculated to be 0.226 ± 0.072 mg and 0.430 ± 0.048 mg in passive and iontophoretic delivery, respectively. Diclofenac was not detected in the subcutaneous tissue after intravenous administration of 1.5 mg/kg diclofenac solution. The trend indicates that WEDD(®) can be used to successfully deliver diclofenac to subcutaneous tissue to concentrations higher when compared to either passive delivery or intravenous dosing of 1.5 mg/kg.

Neural regulation of inflammation: no neural connection from the vagus to splenic sympathetic neurons.

The 'inflammatory reflex' acts through efferent neural connections from the central nervous system to lymphoid organs, particularly the spleen, that suppress the production of inflammatory cytokines. Stimulation of the efferent vagus has been shown to suppress inflammation in a manner dependent on the spleen and splenic nerves. The vagus does not innervate the spleen, so a synaptic connection from vagal preganglionic neurons to splenic sympathetic postganglionic neurons was suggested. We tested this idea in rats. In a preparatory operation, the anterograde tracer DiI was injected bilaterally into the dorsal motor nucleus of vagus and the retrograde tracer Fast Blue was injected into the spleen. On histological analysis 7-9 weeks later, 883 neurons were retrogradely labelled from the spleen with Fast Blue as follows: 89% in the suprarenal ganglia (65% left, 24% right); 11% in the left coeliac ganglion; but none in the right coeliac or either of the superior mesenteric ganglia. Vagal terminals anterogradely labelled with DiI were common in the coeliac but sparse in the suprarenal ganglia, and confocal analysis revealed no putative synaptic connection with any Fast Blue-labelled cell in either ganglion. Electrophysiological experiments in anaesthetized rats revealed no effect of vagal efferent stimulation on splenic nerve activity or on that of 15 single splenic-projecting neurons recorded in the suprarenal ganglion. Together, these findings indicate that vagal efferent neurons in the rat neither synapse with splenic sympathetic neurons nor drive their ongoing activity.

Analysis of in situ manganese(II) oxidation in the Columbia River and offshore plume: linking Aurantimonas and the associated microbial community to an active biogeochemical cycle.

The Columbia River is a major source of dissolved nutrients and trace metals for the west coast of North America. A large proportion of these nutrients are sourced from the Columbia River Estuary, where coastal and terrestrial waters mix and resuspend particulate matter within the water column. As estuarine water is discharged off the coast, it transports the particulate matter, dissolved nutrients and microorganisms forming nutrient-rich and metabolically dynamic plumes. In this study, bacterial manganese oxidation within the plume and estuary was investigated during spring and neap tides. The microbial community proteome was fractionated and assayed for Mn oxidation activity. Proteins from the outer membrane and the loosely bound outer membrane fractions were separated using size exclusion chromatography and Mn(II)-oxidizing eluates were analysed with tandem mass spectrometry to identify potential Mn oxidase protein targets. Multi-copper oxidase (MCO) and haem-peroxidase enzymes were identified in active fractions. T-RFLP profiles and cluster analysis indicates that organisms and bacterial communities capable of oxidizing Mn(II) can be sourced from the Columbia River estuary and nearshore coastal ocean. These organisms are producing up to 10 fM MnO2 cell⁻¹ day⁻¹. Evidence for the presence of Mn(II)-oxidizing bacterial isolates from the genera Aurantimonas, Rhodobacter, Bacillus and Shewanella was found in T-RFLP profiles. Specific Q-PCR probes were designed to target potential homologues of the Aurantimonas manganese oxidizing peroxidase (Mop). By comparing total Mop homologues, Aurantimonas SSU rRNA and total bacterial SSU rRNA gene copies, it appears that Aurantimonas can only account for ~1.7% of the peroxidase genes quantified. Under the broad assumption that at least some of the peroxidase homologues quantified are involved in manganese oxidation, it is possible that other organisms oxidize manganese via peroxidases.

The relationship of the birth date of rat sympathetic neurons to the target they innervate.

In many parts of the nervous system, neurons with the same function often have similar "birth dates" (the time their precursor withdrew from the cell cycle). We investigated the birth dates of eight functional classes of rat sympathetic postganglionic neurons by injecting bromodeoxyuridine during embryonic development, while retrograde tracing and immunohistochemistry were used to identify postganglionic neurons of different functional classes in the mature animals. The times of withdrawal from the cell cycle overlapped, but there were significant differences in the peak time of withdrawal for most of the classes. Furthermore, sympathetic cholinergic postganglionic neurons had a significantly greater proportion of their total population labelled with bromodeoxyuridine than did any of the noradrenergic classes of neurons, indicating prenatal class-specific differences in the handling of bromodeoxyuridine. Together, our findings indicate that, prior to extending axons to their targets, different functional classes of sympathetic neurons show differences in phenotype.

The neurochemistry and innervation patterns of extrinsic sensory and sympathetic nerves in the myenteric plexus of the C57Bl6 mouse jejunum.

In vitro anterograde tracing of axons in mesenteric nerve trunks using biotinamide in combination with immunohistochemical labelling was used to characterize the extrinsic nerve projections in the myenteric plexus of the mouse jejunum. Anterogradely-labelled spinal sensory fibres innervating the enteric nervous system were identified by their immunoreactivity for calcitonin gene-related peptide (CGRP), while sympathetic noradrenergic fibres were detected with tyrosine hydroxylase (TH), using confocal microscopy. The presence of these markers has been previously described in the spinal sensory and sympathetic fibres. Labelled extrinsic nerve fibres in the myenteric plexus were identified apposing enteric neurons that were immunoreactive for either calretinin (CalR), calbindin (CalB) or nitric oxide synthase (NOS). Of the total anterogradely labelled axons in the myenteric plexus, 20% were CGRP-immunoreactive. Labelled CGRP-immunoreactive varicosities were closely apposed to CalR-immunoreactive myenteric cells, many of which were Dogiel type I (40%; interneurons) or type II (20%; intrinsic sensory) neurons. Labelled CGRP-immunoreactive varicosities were also observed in close appositions to CalB-immunoreactive myenteric cell bodies, of which a small subset had type II morphology (18%; intrinsic sensory neurons). A further 43% of all biotinamide-filled fibres were immunoreactive for TH and these fibres were apposed to CalR-immunoreactive cell bodies (small-sized; excitatory motor neurons) and NOS-immunoreactive cell bodies (either type I or small neurons; inhibitory motor neurons and interneurons) in the myenteric plexus. The results provide a neurochemical and neuroanatomical basis for connections between dorsal root afferent neurons and myenteric neurons and suggest an anatomical substrate for the well-known modulation of enteric circuits from sympathetic nerves. No anterogradely-labelled fibres were stained for NOS-immunoreactivity, despite more than 60% of dorsal root ganglion (DRG) neurons retrogradely labelled from the jejunum showing NOS-immunoreactivity. This was due to a substantial, time-dependent, and apparently selective, loss of NOS from extrinsic axons under in vitro conditions. Lastly, a small population of non-immunoreactive biotinamide-filled fibres (<1%) gave rise to dense terminal structures around individual myenteric cell bodies lacking CalR, CalB or NOS. These specialized endings may represent vagal fibres or a subset of spinal sensory neurons that do not contain CGRP.

Generating diversity: Mechanisms regulating the differentiation of autonomic neuron phenotypes.

Sympathetic and parasympathetic postganglionic neurons innervate a wide range of target tissues. The subpopulation of neurons innervating each target tissue can express unique combinations of neurotransmitters, neuropeptides, ion channels and receptors, which together comprise the chemical phenotype of the neurons. The target-specific chemical phenotype shown by autonomic postganglionic neurons arises during development. In this review, we examine the different mechanisms that generate such a diversity of neuronal phenotypes from the pool of apparently homogenous neural crest progenitor cells that form the sympathetic ganglia. There is evidence that the final chemical phenotype of autonomic postganglionic neurons is generated by both signals at the level of the cell body that trigger cell-autonomous programs, as well as signals from the target tissues they innervate.

Aurantimonas manganoxydans, sp. nov. and Aurantimonas litoralis, sp. nov.: Mn(II) oxidizing representatives of a globally distributed clade of alpha-Proteobacteria from the order Rhizobiales.

Several closely related Mn(II)-oxidizing alpha-Proteobacteria were isolated from very different marine environments: strain SI85-9A1 from the oxic/anoxic interface of a stratified Canadian fjord, strain HTCC 2156 from the surface waters off the Oregon coast, and strain AE01 from the dorsal surface of a hydrothermal vent tubeworm. 16S rRNA analysis reveals that these isolates are part of a tight phylogenetic cluster with previously characterized members of the genus Aurantimonas. Other organisms within this clade have been isolated from disparate environments such as surface waters of the Arctic and Mediterranean seas, a deep-sea hydrothermal plume, and a Caribbean coral. Further analysis of all these strains revealed that many of them are capable of oxidizing dissolved Mn(II) and producing particulate Mn(III/IV) oxides. Strains SI85-9A1 and HTCC 2156 were characterized further. Despite sharing nearly identical 16S rRNA gene sequences with the previously described Aurantimonas coralicida, whole genome DNA-DNA hybridization indicated that their overall genomic similarity is low. Polyphasic phenotype characterization further supported distinguishing characteristics among these bacteria. Thus SI85-9A1 and HTCC 2156 are described as two new species within the family 'Aurantimionadaceae': Aurantimonas manganoxydans sp. nov. and Aurantimonas litoralis sp. nov. This clade of bacteria is widely distributed around the globe and may be important contributors to Mn cycling in many environments. Our results highlight the difficulty in utilizing 16S rRNA-based approaches to investigate the microbial ecology of Mn(II) oxidation.

Neurochemical and morphological phenotypes of vagal afferent neurons innervating the adult mouse jejunum.

Whilst much is known about the function and influence of vagal afferents on the mammalian upper gastrointestinal tract, the phenotypes of the different types of vagal afferent neurons innervating the jejunum is unknown. We have previously shown that spinal afferents supplying the jejunum are predominantly medium-sized sensory neurons that express specific combinations of transient receptor potential vanilloid type 1 (TRPV1), neuronal nitric oxide synthase (NOS) and calcitonin-gene related peptide (CGRP) and that they lack binding for isolectin B4 (IB4). This study aimed to identify the chemical phenotypes and somal sizes of jejunal afferent neurons in the mouse vagal ganglion. Jejunal vagal afferents were identified by retrograde labelling with sub-serosal injections of cholera toxin B (CTB) into the jejunal wall and assessed for IB4-binding, TRPV1-, NOS- and CGRP-immunoreactivities using fluorescent microscopy. Almost all (99%) of CTB-labelled vagal afferent neurons were small- and medium-sized sensory cells. Most (81%) jejunal vagal afferents bound IB4 but fewer (32%) expressed TRPV1. A quarter (25%) of those that bound IB4 co-expressed TRPV1-immunoreactivity whilst 77% of TRPV1-expressing jejunal vagal afferent neurons bound IB4. NOS (0%) and CGRP (0%) expression was absent from all CTB-labelled cells examined. In conclusion, vagal afferents innervating the jejunum differ in their expression of IB4, TRPV1, CGRP and NOS from their spinal counterparts, suggesting that the peripheral endings for extrinsic sensory neurons terminating within the enteric nervous system can be identified selectively.

Mn(II) oxidation is catalyzed by heme peroxidases in "Aurantimonas manganoxydans" strain SI85-9A1 and Erythrobacter sp. strain SD-21.

A new type of manganese-oxidizing enzyme has been identified in two alphaproteobacteria, "Aurantimonas manganoxydans" strain SI85-9A1 and Erythrobacter sp. strain SD-21. These proteins were identified by tandem mass spectrometry of manganese-oxidizing bands visualized by native polyacrylamide gel electrophoresis in-gel activity assays and fast protein liquid chromatography-purified proteins. Proteins of both alphaproteobacteria contain animal heme peroxidase and hemolysin-type calcium binding domains, with the 350-kDa active Mn-oxidizing protein of A. manganoxydans containing stainable heme. The addition of both Ca(2+) ions and H(2)O(2) to the enriched protein from Aurantimonas increased manganese oxidation activity 5.9-fold, and the highest activity recorded was 700 microM min(-1) mg(-1). Mn(II) is oxidized to Mn(IV) via an Mn(III) intermediate, which is consistent with known manganese peroxidase activity in fungi. The Mn-oxidizing protein in Erythrobacter sp. strain SD-21 is 225 kDa and contains only one peroxidase domain with strong homology to the first 2,000 amino acids of the peroxidase protein from A. manganoxydans. The heme peroxidase has tentatively been named MopA (manganese-oxidizing peroxidase) and sheds new light on the molecular mechanism of Mn oxidation in prokaryotes.

C1 neurons in the rat rostral ventrolateral medulla differentially express vesicular monoamine transporter 2 in soma and axonal compartments.

Vesicular monoamine transporter 2 (VMAT2) packages biogenic amines into large dense core and synaptic vesicles for either somatodendritic or synaptic release from neurons of the CNS. Whilst the distribution of VMAT2 has been well characterized in many catecholaminergic cell groups, its localization amongst C1 adrenergic neurons in the medulla has not been examined in detail. Within the rostral ventrolateral medulla (RVLM), C1 neurons are a group of barosensitive, adrenergic neurons. Rostral C1 cells project to the thoracic spinal cord and are considered sympathetic premotor neurons. The majority of caudal C1 cells project rostrally to regions such as the hypothalamus. The present study sought to quantitate the somatodendritic expression of VMAT2 in C1 neurons, and to assess the subcellular distribution of the transporter. Immunoreactivity for VMAT2 occurred in 31% of C1 soma, with a high proportion of these in the caudal part of the RVLM. Retrograde tracing studies revealed that only two of 43 bulbospinal C1 neurons contained faint VMAT2-immunoreactivity, whilst 88 +/- 5% of rostrally projecting neurons were VMAT2-positive. A lentivirus, designed to express green fluorescent protein exclusively in noradrenergic and adrenergic neurons, was injected into the RVLM to label C1 neurons. Eighty-three percent of C1 efferents that occurred in close proximity to sympathetic preganglionic neurons within the T(3) intermediolateral cell column contained VMAT2-immunoreactivity. These data demonstrate differential distribution of VMAT2 within different subpopulations of C1 neurons and suggest that this might reflect differences in somatodendritic vs. synaptic release of catecholamines.

Distinct chemical classes of medium-sized transient receptor potential channel vanilloid 1-immunoreactive dorsal root ganglion neurons innervate the adult mouse jejunum and colon.

Physiological studies suggest visceral spinal afferents are generally small diameter, unmyelinated C-fibers or myelinated Adelta-fibers, but little is known about the size and chemical phenotypes of visceral sensory neurons supplying the small intestine. This study examines the size and expression patterns of transient receptor potential vanilloid 1 (TRPV1), calcitonin gene-related peptide (CGRP), substance P (SP), neuronal nitric oxide synthase (NOS) and isolectin B4-binding (IB4) in dorsal root ganglion (DRG) neurons projecting to the gastrointestinal tract. The spinal afferent innervation of mouse jejunum and distal colon was investigated with retrograde neuronal tracing and multi-label immunohistochemistry. Expression of histochemical markers and soma sizes of retrogradely labeled DRG profiles were determined with confocal microscopy. Most (>75%) jejunal and colonic afferent neurons were medium- and large-sized cells. The majority (82%) of jejunal afferents expressed TRPV1, but few bound IB4. All retrogradely labeled jejunal afferents expressing NOS-immunoreactivity (64%) also expressed TRPV1 and CGRP and most expressed SP. Most labeled colonic afferents expressed TRPV1 (62%) and half expressed NOS. Taken together these data demonstrate that the spinal afferent supply of the jejunum and colon is largely from medium and large sensory neurons, suggesting most intestinal afferent axons are A fibers. The various chemically-defined subpopulations of afferents may play multiple roles in sensory innervation of the jejunum apart from nociceptive transduction. Additionally, we have identified a unique chemical code, TRPV1/NOS/CGRP/SP, that distinguishes many spinal afferent terminals from those of enteric neurons.