Advances in colonic motor complexes in mice.

The primary functions of the gastrointestinal (GI) tract are to absorb nutrients, water, and electrolytes that are essential for life. This is accompanied by the capability of the GI tract to mix ingested content to maximize absorption and effectively excrete waste material. There have been major advances in understanding intrinsic neural mechanisms involved in GI motility. This review highlights major advances over the past few decades in our understanding of colonic motor complexes (CMCs), the major intrinsic neural patterns that control GI motility. CMCs are generated by rhythmic coordinated firing of large populations of myenteric neurons. Initially, it was thought that serotonin release from the mucosa was required for CMC generation. However, careful experiments have now shown that neither the mucosa nor endogenous serotonin are required, although, evidence suggests enteroendocrine (EC) cells modulate CMCs. The frequency and extent of propagation of CMCs are highly dependent on mechanical stimuli (circumferential stretch). In summary, the isolated mouse colon emerges as a good model to investigate intrinsic mechanisms underlying colonic motility and provides an excellent preparation to explore potential therapeutic agents on colonic motility, in a highly controlled in vitro environment. In addition, during CMCs, the mouse colon facilitates investigations into the emergence of dynamic assemblies of extensive neural networks, applicable to the nervous system of different organisms.

Land-Atmosphere Responses to a Total Solar Eclipse in Three Ecosystems With Contrasting Structure and Physiology.

Mid-Missouri experienced up to 2 min 40 s of totality at around solar noon during the total eclipse of 2017. We conducted the Mid-Missouri Eclipse Meteorology Experiment to examine land-atmosphere interactions during the eclipse. Here, research examining the eclipse responses in three contrasting ecosystems (forest, prairie, and soybeans) is described. There was variable cloudiness around first and fourth contacts (i.e., the start and end of partial solar obscuration) at the forest and prairie; however, solar irradiance (K ↓) signals during the eclipse were relatively clean. Unfortunately, the eclipse forcing at the soybean field was contaminated by convective activity, which decreased K ↓ beginning about an hour before first contact and exposed the field to cold outflow ~30 min before second contact. Turbulence was suppressed during the eclipse at all sites; however, there was also an amplified signal at the soybean field during the passage of a gust front. The standard deviations of the horizontal and vertical wind velocities and friction velocities decreased by ~75% at the forest (aerodynamically rough), and ~60% at the prairie (aerodynamically smooth). The eddy fluxes of energy were highly coherent with the solar forcing with the latent and sensible heat fluxes approaching 0 W/m2 and changing in direction, respectively. For the prairie site, we estimated a canopy-scale time constant for the surface conductance light response of 10 min. Although the eclipse imparted large forcings on surface energy balances, the air temperature response was relatively muted (1.5-2.5 °C decrease) due to the absence of topographic effects and the relatively moist land and atmosphere.

OCO-2 advances photosynthesis observation from space via solar-induced chlorophyll fluorescence.

Quantifying gross primary production (GPP) remains a major challenge in global carbon cycle research. Spaceborne monitoring of solar-induced chlorophyll fluorescence (SIF), an integrative photosynthetic signal of molecular origin, can assist in terrestrial GPP monitoring. However, the extent to which SIF tracks spatiotemporal variations in GPP remains unresolved. Orbiting Carbon Observatory-2 (OCO-2)'s SIF data acquisition and fine spatial resolution permit direct validation against ground and airborne observations. Empirical orthogonal function analysis shows consistent spatiotemporal correspondence between OCO-2 SIF and GPP globally. A linear SIF-GPP relationship is also obtained at eddy-flux sites covering diverse biomes, setting the stage for future investigations of the robustness of such a relationship across more biomes. Our findings support the central importance of high-quality satellite SIF for studying terrestrial carbon cycle dynamics.

Systemic gene delivery transduces the enteric nervous system of guinea pigs and cynomolgus macaques.

Characterization of adeno-associated viral vector (AAV) mediated gene delivery to the enteric nervous system (ENS) was recently described in mice and rats. In these proof-of-concept experiments, we show that intravenous injections of clinically relevant AAVs can transduce the ENS in guinea pigs and non-human primates. Neonatal guinea pigs were given intravenous injections of either AAV8 or AAV9 vectors that contained a green fluorescent protein (GFP) expression cassette or phosphate-buffered saline. Piglets were euthanized three weeks post injection and tissues were harvested for immunofluorescent analysis. GFP expression was detected in myenteric and submucosal neurons along the length of the gastrointestinal tract in AAV8 injected guinea pigs. GFP-positive neurons were found in dorsal motor nucleus of the vagus and dorsal root ganglia. Less transduction occurred in AAV9-treated tissues. Gastrointestinal tissues were analyzed from young cynomolgus macaques that received systemic injection of AAV9 GFP. GFP expression was detected in myenteric neurons of the stomach, small and large intestine. These data demonstrate that ENS gene delivery translates to larger species. This work develops tools for the field of neurogastroenterology to explore gut physiology and anatomy using emerging technologies such as optogenetics and gene editing. It also provides a basis to develop novel therapies for chronic gut disorders.

Dairy Manure Total Solid Levels Impact CH Flux and Abundance of Methanogenic Archaeal Communities.

Stored liquid dairy manures are methane (CH) emission hotspots because of the large amount of slurry volatile solids (VS) converted into CH by methanogens under anaerobic conditions. Our research has indicated that a reduction of total solids (TS) of slurries before storage can reduce CH emissions. In the current study, methanogen abundance was characterized in tanks with different CH emissions. Using mesoscale slurry storage facilities equipped for continuous gaseous emission monitoring, we stored dairy slurries having TS from 9.5 to 0.3% for up to 6 mo. Samples were taken after Day 30 and Day 120 of the storage (20 May-16 Nov. 2010) from the upper and bottom layers of the slurries. Methanogenic communities were studied by targeting the gene encoding the α subunit methyl-coenzyme M reductase (), which catalyzes the final step of methanogenesis. Interestingly, mean abundances of methanogens increased by ∼8 and 23% at the top and bottom sections, respectively, as slurry TS decreased from 9.5 to 0.3%. Cumulative CH emissions, however, decreased by ∼70% as slurry TS decreased from 9.5 to 0.3%. Nevertheless, compared with Day 30 of storage, mean abundances of methanogens were relatively higher at Day 120 (up to 19%), consistent with an increase in the cumulative CH emissions. Polymerase chain reaction denaturing gel electrophoresis analysis indicated a low methanogen diversity, with most bands sequenced closely related to the genus (>95% amino acid sequence similarity), the hydrogenotrophic methanogens. Results suggest that available carbon substrate and not methanogen abundance may be limiting cumulative CH emissions at reduced TS levels of dairy slurries.

Deficiency in the mRNA export mediator Gle1 impairs Schwann cell development in the zebrafish embryo.

GLE1 mutations cause lethal congenital contracture syndrome 1 (LCCS1), a severe autosomal recessive fetal motor neuron disease, and more recently have been associated with amyotrophic lateral sclerosis (ALS). The gene encodes a highly conserved protein with an essential role in mRNA export. The mechanism linking Gle1 function to motor neuron degeneration in humans has not been elucidated, but increasing evidence implicates abnormal RNA processing as a key event in the pathogenesis of several motor neuron diseases. Homozygous gle1(-/-) mutant zebrafish display various aspects of LCCS, showing severe developmental abnormalities including motor neuron arborization defects and embryonic lethality. A previous gene expression study on spinal cord from LCCS fetuses indicated that oligodendrocyte dysfunction may be an important factor in LCCS. We therefore set out to investigate the development of myelinating glia in gle1(-/-) mutant zebrafish embryos. While expression of myelin basic protein (mbp) in hindbrain oligodendrocytes appeared relatively normal, our studies revealed a prominent defect in Schwann cell precursor proliferation and differentiation in the posterior lateral line nerve. Other genes mutated in LCCS have important roles in Schwann cell development, thereby suggesting that Schwann cell deficits may be a common factor in LCCS pathogenesis. These findings illustrate the potential importance of glial cells such as myelinating Schwann cells in motor neuron diseases linked to RNA processing defects.

Endogenous CRF in rat large intestine mediates motor and secretory responses to stress.

BACKGROUND: Corticotropin-releasing factor (CRF) mediates our body's overall responses to stress. The role of central CRF in stress-stimulated colonic motility is well characterized. We hypothesized that transient perturbation in expression of enteric CRF is sufficient to change stress-induced colonic motor and secretory responses. METHODS: Sprague-Dawley rats (adult, male) were subjected to 1-h partial restraint stress (PRS) and euthanized at 0, 4, 8, and 24 h. CRF mRNA and peptide levels in the colon were quantified by real-time RT-PCR, enzyme immuno-assay and immunohistochemistry. Double-stranded RNA (dsRNA) designed to target CRF (dsCRF) was injected into the colonic wall to attain RNA interference-mediated inhibition of CRF mRNA expression. DsRNA for ß-globin was used as a control (dsControl). Four days after dsRNA injection, rats were subjected to 1-h PRS. Fecal output was measured. Ussing chamber techniques were used to assess colonic mucosal ion secretion and transepithelial tissue conductance. KEY RESULTS: Exposure to PRS elevated CRF expression and increased CRF release in the rat colon. Injection of dsCRF inhibited basal CRF expression and prevented the PRS-induced increase in CRF expression, whereas CRF expression in dsControl-injected colons remained high after PRS. In rats treated with dsControl, PRS caused a significant increase in fecal pellet output, colonic baseline ion secretion, and transepithelial tissue conductance. Inhibition of CRF expression in the colon prevented PRS-induced increase in fecal output, baseline ion secretion, and transepithelial tissue conductance. CONCLUSIONS & INFERENCES: These results provide direct evidence that transient perturbation in peripherally expressed CRF prevents colonic responses to stress.

Silage effluent management: a review.

Silage effluent is a potent wastewater that can be produced when ensiling crops that have a high moisture content (MC). Silage effluent can cause fish-kills and eutrophication due to its high biochemical oxygen demand (BOD) and nutrient content, respectively. It has a high acidity (pH ≈ 3.5-5) making it corrosive to steel and damaging to concrete, which makes handling, storage and disposal a challenge. Although being recognized as a concentrated wastewater, most research has focused on preventing its production. Despite noted imprecision in effluent production models-and therefore limited ability to predict when effluent will flow-there has been little research aimed at identifying effective reactive management options, such as containment and natural treatment systems. Increasing climate variability and intensifying livestock agriculture are issues that will place a greater importance on developing comprehensive, multi-layered management strategies that include both preventative and reactive measures. This paper reviews important factors governing the production of effluent, approaches to minimize effluent flows as well as treatment and disposal options. The challenges of managing silage effluent are reviewed in the context of its chemical constituents. A multi-faceted approach should be utilized to minimize environmental risks associated with silage effluent. This includes: (i) managing crop moisture content prior to ensiling to reduce effluent production, (ii) ensuring the integrity of silos and effluent storages, and (iii) establishing infrastructure for effluent treatment and disposal. A more thorough investigation of constructed wetlands and vegetated infiltration areas for treating dilute silage effluent is needed. In particular, there should be efforts to improve natural treatment system design criteria by identifying pre-treatment processes and appropriate effluent loading rates. There is also a need for research aimed at understanding the effects of repeated land application of effluent on soil quality and crop yields, as spreading is a common disposal practice.

Effects of low protein diets on pigs with a lean genotype 2. Compositional traits measured with computed tomography (CT).

The effects on compositional changes across the growing-finishing period (40-115 kg) of feeding pig diets with different protein and amino acid levels were investigated using CT scanning (at 60, 85 and 115 kg live weight). Pigs of a lean commercial genotype were fed a commercial control regime (C), or a low protein regime with either high (LP1) or low (LP2) essential amino acid levels, all balanced for net energy. In vivo CT measurements agreed well with post-slaughter sample joint dissection results for carcass tissue weights/proportions, and CT-measured muscle density predicted intramuscular fat accurately. Pigs on C and LP1 regimes did not differ significantly in composition during growth. However, pigs on the LP2 regime had significantly more fat (in carcass, internal and intra-muscular depots) and less muscle, from 85 kg onwards. Although fat levels differed depending on diet regime (LP2>others), proportions of fat in different body depots were unaffected.

Effects of low protein diets on pigs with a lean genotype. 1. Carcass composition measured by dissection and muscle fatty acid composition.

Pigs with a lean genotype were fed diets differing in protein and amino acid contents between 40 and 115 kg live weight. A high protein control regime (C) was compared with one supplying 11% less total protein but the same essential amino acid levels (LP1) and one supplying 16% less protein but lower amino acid levels (LP2). Regime LP2 produced fatter pigs in terms of subcutaneous, intermuscular and intramuscular fat (IMF), the latter measured in longissimus and semimembranosus muscles. The percentage of linoleic acid was lowest and that of oleic acid highest in IMF from LP2 pigs (11.57 and 34.59% respectively in longissimus). Pigs in regime LP1 had more longissimus IMF than C but similar semimembranosus IMF although both muscles had lower percentages of linoleic acid in LP1 than C, suggesting a tendency towards greater fat deposition in LP1. The high IMF content in LP2 produced the most tender, juicy steaks.

Neurogenic mucosal bicarbonate secretion in guinea pig duodenum.

BACKGROUND AND PURPOSE: To test a hypothesis that: (i) duodenal pH and osmolarity are individually controlled at constant set points by negative feedback control centred in the enteric nervous system (ENS); (ii) the purinergic P2Y(1) receptor subtype is expressed by non-cholinergic secretomotor/vasodilator neurons, which represent the final common excitatory pathway from the ENS to the bicarbonate secretory glands. EXPERIMENTAL APPROACH: Ussing chamber and pH-stat methods investigated involvement of the P2Y(1) receptor in neurogenic stimulation of mucosal bicarbonate (HCO(3)(-)) secretion in guinea pig duodenum. KEY RESULTS: ATP increased HCO(3)(-) secretion with an EC(50) of 160 nM. MRS2179, a selective P2Y(1) purinergic receptor antagonist, suppressed ATP-evoked HCO(3)(-) secretion by 47% and Cl(-) secretion by 63%. Enteric neuronal blockade by tetrodotoxin or exposure to a selective vasoactive intestinal peptide (VIP, VPAC(1)) receptor antagonist suppressed ATP-evoked HCO(3)(-) secretion by 61 and 41%, respectively, and Cl- by 97 and 70% respectively. Pretreatment with the muscarinic antagonist, scopolamine did not alter ATP-evoked HCO3(-) or Cl(-) secretion. CONCLUSION AND IMPLICATIONS: Whereas acid directly stimulates the mucosa to release ATP and stimulate HCO(3)(-) secretion in a cytoprotective manner, neurogenically evoked HCO(3)(-) secretion accounts for feedback control of optimal luminal pH for digestion. ATP stimulates duodenal HCO(3)(-) secretion through an excitatory action at purinergic P2Y(1) receptors on neurons in the submucosal division of the ENS. Stimulation of the VIPergic non-cholinergic secretomotor/vasodilator neurons, which are one of three classes of secretomotor neurons, accounts for most, if not all, of the neurogenic secretory response evoked by ATP.

Relationships between dairy slurry total solids, gas emissions, and surface crusts.

Livestock slurry storages are sources of methane (CH4), nitrous oxide (NO2), and ammonia (NH3) emissions. Total solids (TS) content is an indicator of substrate availability for CH4 and N2O production and NH3 emissions and is related to crust formation, which can affect these gas emissions. The effect of TS on these emissions from pilot-scale slurry storages was quantified from 20 May through 16 Nov. 2010 in Nova Scotia, Canada. Emissions from six dairy slurries with TS ranging from 0.3 to 9.5% were continuously measured using flow-through steady-state chambers. Methane emissions modeled using the USEPA methodology were compared with measured data focusing on emissions when empty storages were filled, and retention times were >30 d with undegraded volatile solids (VS) remaining in the system considered available for CH4 production (VS carry-over). Surface crusts formed on all the slurries. Only the slurries with TS of 3.2 and 5.8% were covered completely for ∼3 mo. Nitrous oxide contributed <5% of total greenhouse gas emissions for all TS levels. Ammonia and CH4 emissions increased linearly with TS despite variable crusting, suggesting substrate availability for gas production was more important than crust formation in regulating emissions over long-term storage. Modeled CH4 emissions were substantially higher than measured data in the first month, and accounting for this could improve overall model performance. Carried-over VS were a CH4 source in months 2 through 6. The results of this study suggest that substrate availability regulates emissions over long-term storage and that modifying the USEPA model to better describe carbon cycling is warranted.

Effect of dietary chicory on boar taint.

Following preliminary screening and feeding trials on farms supplying a commercial abattoir, 360 entire male pigs were used to evaluate the effects of different percentages of chicory (Cichorium intybus L.) on levels of boar taint compounds and sensory aspects in backfat. Pigs were fed 0, 3, 6 or 9% chicory in the diet, 30 pigs being sampled at 3 different times: initially to measure basal levels of skatole and androstenone and after 1 and 2 weeks on the test diets. Cooked samples of backfat were presented to a trained sensory panel for "sniff" tests. Chicory fed at 9% for 2 weeks reduced skatole levels significantly (P<0.001), with 0.55 of pigs below 0.05 µg/g, typical of levels in castrated males. Abnormal odour scores were significantly lower for pigs in this group compared with 0% pigs (P<0.001), however, androstenone concentration was significantly higher in this group after the 2 week feeding period (P<0.005). Thus, feeding 9% chicory for 2 weeks was effective in reducing backfat skatole concentrations and abnormal odour scores of cooked fat but not androstenone concentration.

Nonruminant Nutrition Symposium: Neurogastroenterology and food allergies.

Neurogastroenterology is a subspecialty encompassing relations of the nervous system to the gastrointestinal tract. The central concept is emergence of whole organ behavior from coordinated activity of the musculature, mucosal epithelium, and blood vasculature. Behavior of each effector is determined by the enteric nervous system (ENS). The ENS is a minibrain positioned close to the effectors it controls. The ENS neurophysiology is in the framework of neurogastroenterology. The digestive tract is recognized as the largest lymphoid organ in the body with a unique complement of mast cells. In its position at the "dirtiest" of interfaces between the body and outside world, the mucosal immune system encounters food antigens, bacteria, parasites, viruses, and toxins. Epithelial barriers are insufficient to exclude fully the antigenic load, thereby allowing chronic challenges to the immune system. Observations in antigen-sensitized animals document direct communication between the mucosal immune system and ENS. Communication is functional and results in adaptive responses to circumstances within the lumen that are threatening to the functional integrity of the whole animal. Communication is paracrine and incorporates specialized sensing functions of mast cells for specific antigens together with the capacity of the ENS for intelligent interpretation of the signals. Immuno-neural integration progresses sequentially, beginning with immune detection, followed by signal transfer to the ENS, followed by neural interpretation and then selection of a neural program with coordinated mucosal secretion and a propulsive motor event that quickly clears the threat from the intestinal lumen. Operation of the defense program evokes symptoms of cramping abdominal pain, fecal urgency, and acute watery diarrhea. Investigative approaches to immuno-ENS interactions merge the disciplines of mucosal immunology and ENS neurophysiology into the realm of neurogastroenterology.

Effect of vitamin E supplementation and diet on fatty acid composition and on meat colour and lipid oxidation of lamb leg steaks displayed in modified atmosphere packs.

Groups of 8 lambs were allocated to one of five concentrate diets supplemented with all-rac-α-tocopheryl acetate containing 30 (C30), 60 (C60), 120 (C120), 250 (C250) and 500 (C500) mg/kg dry matter. Two other groups were fed grass silage and 400 g/day concentrate with 60 (S60) or 500 (S500) mg α-tocopheryl acetate/kg dry matter. Within diet, vitamin E level did not affect growth performance or carcass characteristics. Basal diet did not affect final live weight, conformation and fatness scores. M. semimembranosus from S lambs contained more α-tocopherol than that of C lambs on the same intake and by day 6 in MAP (75%O2/25%CO2) chroma and a* were below acceptable levels in C30 lambs. TBARS were higher in C30 and C60 muscle than in other treatments (P<0.001) after 3 and 6 days display. Muscle fatty acid composition varied with basal diet but lipid oxidation depended more on vitamin E concentration with an initial concentration of 1.9 µg/g muscle preventing significant lipid oxidation.

Inducing electronic changes in graphene through silicon (100) substrate modification.

We have performed scanning tunneling microscopy and spectroscopy (STM/STS) measurements as well as ab initio calculations for graphene monolayers on clean and hydrogen(H)-passivated silicon (100) (Si(100)/H) surfaces. In order to experimentally study the same graphene piece on both substrates, we develop a method to depassivate hydrogen from under graphene monolayers on the Si(100)/H surface. Our work represents the first demonstration of successful and reproducible depassivation of hydrogen from beneath monolayer graphene flakes on Si(100)/H by electron-stimulated desorption. Ab initio simulations combined with STS taken before and after hydrogen desorption demonstrate that graphene interacts differently with the clean and H-passivated Si(100) surfaces. The Si(100)/H surface does not perturb the electronic properties of graphene, whereas the interaction between the clean Si(100) surface and graphene changes the electronic states of graphene significantly. This effect results from the covalent bonding between C and surface Si atoms, modifying the π-orbital network of the graphene layer. The local density of states shows that the bonded C and Si surface states are highly disturbed near the Fermi energy.

Comparison of heating methods and the use of different tissues for sensory assessment of abnormal odours (boar taint) in pig meat.

Five heating methods (microwave, hotwire, boiling at 25 °C and 75 °C and melting) were used to generate cooking odours from backfat of entire male pigs and a 'composite' sample consisting of fat and muscle from the head along with salivary glands. The methods elicited significantly different scores for odours from 4 groups of 10 samples differing in their concentrations and ratios of skatole and androstenone. The odours (pork odour, abnormal odour, skatole odour and androstenone odour) were assessed by 3 experienced assessors. Correlations between skatole and androstenone concentrations and abnormal odour score in backfat were higher for skatole, suggesting it is the more important boar taint compound. In the composite sample, androstenone concentration was much higher than in backfat and androstenone was a more important contributor to boar taint. The microwave, hotwire and boiling (75 °C) methods produced the clearest separation between samples and the microwave method was considered the most suitable for on-line use.

Population dynamics of Escherichia coli inoculated by irrigation into the phyllosphere of spinach grown under commercial production conditions.

Recent outbreaks of food-borne illnesses associated with the consumption of fresh produce have increased attention on irrigation water as a potential source of pathogen contamination. A better understanding of the behaviour of enteric pathogens introduced into agricultural systems during irrigation will aid in risk assessments and support the development of appropriate farm-level water management practices. For this reason, the survival dynamics of two nalidixic acid resistant strains of Escherichia coli after their spray inoculation into the phyllosphere and soil of field spinach were examined over two growing seasons. E. coli strains NAR, an environmental isolate, and DM3n, a non-pathogenic serotype O157:H7, were applied at rates of 104 to 107 cfu/100ml to the fully developed spinach plants that arose subsequent to the harvesting of their upper leafy portions for commercial purposes (secondary-growth plants). After 72 h, E. coli on spinach were reduced by 3-5 logs. Culturable E. coli were recovered from plants up to 6 days post-inoculation. Survival in soil was greater than in the phyllosphere. Under ambient conditions, the mean 72 h first order decay constant computed by Chick's Law was 0.1 h⁻¹. Although light reduction studies indicated UV irradiation negatively influenced the persistence of E. coli, a simple relationship between UV exposure and phyllosphere E. coli densities could not be established. E. coli introduced to the leafy portions of spinach via spray irrigation displayed rapid declines in their culturability under the open environmental conditions experienced during this study. A 6 day period between the last irrigation and harvest would minimize the risks of E. coli survival in the spinach phyllosphere. E. coli NAR was identified as a possible surrogate for the O157:H7 strain, DM3n.

Identification of QTL with effects on fatty acid composition of meat in a Charolais x Holstein cross population.

A whole-genome scan was carried out to detect quantitative trait loci (QTL) influencing beef fatty acid composition using a CharolaisxHolstein population established using a balanced F2 and Backcross breeding design. The phenotypes considered in this study included a total of 24 fatty acid related traits determined in loin muscle samples of the 235 second-generation cross-bred bull calves of the herd. The QTL regression analysis performed, based on 165 microsatellite markers distributed across the 29 bovine autosomes, identified 34 QTL with F-ratios exceeding the 5% chromosome-wide significance threshold. Three of these QTL, one located on chromosome 1 (for the content on linoleic acid, C18:2n-6) and two on chromosome 10 (for the content of gamma-linoleic DPA-docosapentaenoic and DPA-docosapentaenoic, C20:3n-6 and C22:5n-3), also exceeded the 5% genome-wide significance level. A follow-up analysis correcting for intramuscular fat content showed that some of the QTL detected initially (e.g. those localised on chromosome 22) were influenced by fat deposition differences between the founder breeds. The coincident location of some of the linkage associations identified and QTL previously reported for beef fatty acid composition and other meat quality traits, in the same or other cattle populations, provides supporting evidence for the results reported here.

Differential actions of urocortins on neurons of the myenteric division of the enteric nervous system in guinea pig distal colon.

BACKGROUND AND PURPOSE: Urocortins (Ucns) 1, 2 and 3 are corticotropin-releasing factor (CRF)-related neuropeptides and may be involved in neural regulation of colonic motor functions. Nevertheless, details of the neural mechanism of action for Ucns have been unclear. We have, here, tested the hypothesis that Ucns act in the enteric nervous system (ENS) to influence colonic motor behaviour. EXPERIMENTAL APPROACH: We used intracellular recording with 'sharp' microelectrodes, followed by intraneuronal injection of biocytin, and immunohistochemical localization of CRF(1) and CRF(2) receptors in guinea pig colonic tissue. KEY RESULTS: Application of Ucn1 depolarized membrane potentials and elevated excitability in 58% of AH-type and 60% of S-type colonic myenteric neurons. In most of the neurons tested, depolarizing responses evoked by Ucn-1 were suppressed by the CRF(1) receptor antagonist NBI 27914, but were unaffected by the CRF(2) receptor antagonist antisauvagine-30. The selective CRF(2) receptor agonists, Ucn2 and Ucn3, evoked depolarizing responses in 12 and 8% of the AH-type myenteric neurons, respectively, and had no effect on S-type neurons. Antisauvagine-30, but not NBI 27914, suppressed these Ucn2- and Ucn3-evoked responses. Immunohistochemical staining identified CRF(1) as the predominant CRF receptor subtype expressed by ganglion cell somas, while CRF(2)-immunoreactive neuronal somas were sparse. Ucns did not affect excitatory synaptic transmission in the ENS. CONCLUSIONS AND IMPLICATIONS: The results suggest that Ucns act as neuromodulators to influence myenteric neuronal excitability. The excitatory action of Ucn1 in myenteric neurons was primarily at CRF(1) receptors, and the excitatory action of Ucn2 and Ucn3 was at CRF(2) receptors.