Baseline Short-circuit Current Research Articles

Physiological bioactivity of a postbiotic consisting of heat-treated lactobacilli on mouse small intestine

Lactobacillus LB is a postbiotic generated following fermentation by Limosilactobacillus fermentum and Lactobacillus delbrueckii. Lactobacillus LB alleviates acute diarrhoea and ameliorates the symptoms of irritable bowel syndrome. Here, we investigated whether modulation of intestinal ion transport and motility contributes to these beneficial effects and whether the postbiotic produced with both strains contributes to a unique biophysiological profile. In Ussing chamber studies, low lactose-Lactobacillus LB (LL-LB) significantly increased baseline short-circuit current, and this was partially mediated by sodium-D-glucose transporter 1. In organ baths, LL-LB significantly decreased ileal tone and increased carbachol-induced contractility. Relative to LL-LB, preparations produced using a single strain fermentate generated from L. fermentum significantly increased baseline short-circuit current and inhibited carbachol-induced contractility. Our data demonstrate a unique biophysiological profile for the dual strain postbiotic and support a direct and immediate effect of LL-LB on host physiology ex vivo which could contribute to the clinical efficacy of Lactobacillus LB.

Acetylcholine-related proteins in non-neoplastic appearing colonic mucosa from patients with colorectal neoplasia.

The pathogenesis of colorectal neoplasia (CRN) has been associated with altered non-neuronal acetylcholine (ACh) metabolism. The aim of this study was to characterize expression, function, and cellular location of ACh-related proteins in biopsies obtained from endoscopic normal-appearing sigmoid colon in patients with and without CRN. Messenger-RNA (mRNA) levels of 17 ACh-related proteins were quantified by rt-qPCR. Functional responses to ACh, measured as electrogenic transepithelial short circuit current (SCC), were recorded using the Ussing chamber technique. Finally, cellular localization of choline transporter-like proteins (CTLs) and butyryl-cholinesterase enzyme (BChE) was determined by immunohistochemistry. mRNA expression of CTL1 and CTL4 was increased in patients with CRN (P = 0.002 and P = 0.04, respectively). In functional experiments, baseline SCC was increased in CRN patients. ACh induced rapid biphasic changes in SCC. An initial decreasing phase was observed in the minority of CRN patients versus the majority of controls (25% vs 69%, respectively, P = 0.031). For the second increasing phase of SCC, data indicated ACh-activation of two receptors. For both parts of the biphasic response, the half maximal effective concentration and maximal responses showed no difference between patient groups. Immunohistochemistry demonstrated CTL1, 3 and 4 and BChE to be localized to colonic crypt cells. We conclude that CRN is associated with increased expression of CTL1 and CTL4, augmented basal prostaglandin-dependent secretion, and altered functional channel response to ACh in human endoscopic normal-appearing colonic mucosa. The immunohistochemical findings support CTL1, CTL3, CTL4, and BChE to be involved in non-neuronal mucosal ACh metabolism.

Selenium-enriched Agaricus bisporus mushroom protects against increase in gut permeability ex vivo and up-regulates glutathione peroxidase 1 and 2 in hyperthermally-induced oxidative stress in rats.

Dietary effects of organic Se supplementation in the form of Se-enriched Agaricus bisporus mushroom on ileal mucosal permeability and antioxidant selenoenzymes status in heat induced oxidative stress in rats were evaluated. Acute heat stress (40 °C, 21% relative humidity, 90 min exposure) increased ileum baseline short circuit current (Isc; 2.40-fold) and epithelial conductance (Ge; 2.74-fold). Dietary supplementation with Se-enriched A. bisporus (1 µg Se/g feed) reduced (p < 0.05) ileum Isc and Ge during heat stress to 1.74 and 1.91 fold, respectively, indicating protection from heat stress-induced mucosal permeability increase. The expression of ileum glutathione peroxidase (GPx-) 1 and 2 mRNAs were up-regulated (p < 0.05) by 1.90 and 1.87-fold, respectively, for non-heat stress rats on the Se-enriched diet relative to the control. The interplay between heat stress and dietary Se is complex. For rats on the control diet, heat stress alone increased ileum expression of GPx-1 (2.33-fold) and GPx-2 (2.23-fold) relative to thermoneutral conditions. For rats on the Se-enriched diet, heat stress increased (p < 0.05) GPx-1 expression only. Rats on Se-enriched + α-tocopherol diet exhibited increased expression of both genes (p < 0.05). Thus, dietary Se-enriched A. bisporus protected against increase in ileum permeability and up-regulated GPx-1 and GPx-2 expression, selenoenzymes relevant to mitigating oxidative stress.

Up-regulated cystic fibrosis transmembrane conductance regulator after anal stenosis in rats

BackgroundThe pathophysiology of fecal incontinence from fecal impaction and rectal distension is poorly understood. We hypothesize that fecal impaction elicits up-regulation of cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated mucosal chloride channel. MethodsThe anus was ligated to produce 75% stenosis in rats. Controls received ligation without inducing stenosis. 24 to 48 hours after ligation the colon was removed. Mucosal short-circuit current was measured by Ussing chamber. Western blot analysis was used to detect CFTR expression in the colonic mucosa. Ligated rats failed to defecate, whereas control rats stooled normally. ResultsLigated colons were markedly stool filled and dilated. Water content of feces was significantly increased to 66.5% ± 1.1% (P < .01, n = 12) 24 hours after ligation, vs controls (49.5 ± 5.2%, n = 12). Baseline short-circuit current was significantly increased in the distal (78.8 ± 7.4 μA/cm2, n = 8, P < .01) and mid colon (24.5 ± 2.5 μA/cm2, n = 8, P < .05) 24 hours after ligation, compared to control rats (12.5 ± 3.2 μA/cm2, n = 8). CFTR expression was significantly increased 24 hours after ligation in the mid and distal colon. ConclusionWe observe that fecal impaction from anal ligation induces early compensatory up-regulation of CFTR, altering function from net absorption to net secretion in the mid and distal colon.

Early weaning stress impairs development of mucosal barrier function in the porcine intestine

Early life stress is a predisposing factor for the development of chronic intestinal disorders in adult life. Here, we show that stress associated with early weaning in pigs leads to impaired mucosal barrier function. Early weaning (15- to 21-day weaning age) resulted in sustained impairment in intestinal barrier function, as indicated by reductions in jejunal transepithelial electrical resistance and elevations in mucosal-to-serosal flux of paracellular probes [(3)H]mannitol and [(14)C]inulin measured at 5 and 9 wk of age, compared with that shown in late-weaned pigs (23- to 28-day weaning age). Elevated baseline short-circuit current was observed in jejunum from early-weaned pigs and was shown to be mediated via enhanced Cl(-) secretion. Jejunal barrier dysfunction in early-weaned pigs coincided with increased lamina propria immune cell density particularly mucosal mast cells. The mast cell stabilizer drug sodium cromoglycolate ameliorated barrier dysfunction and hypersecretion in early-weaned pigs, demonstrating an important role of mast cells. Furthermore, activation of mast cells ex vivo with c48/80 and corticotrophin-releasing factor (CRF) in pig jejunum mounted in Ussing chambers induced barrier dysfunction and elevations in short-circuit current that were inhibited with mast cell protease inhibitors. Experiments in which selective CRF receptor antagonists were administered to early-weaned pigs revealed that CRF receptor 1 (CRFr1) activation mediates barrier dysfunction and hypersecretion, whereas CRFr2 activation may be responsible for novel protective properties in the porcine intestine in response to early life stress.

Insight into the mechanism of short chain fatty acid absorption in the ostrich (Struthio camelus) proximal colon

1. Occurrence of short chain fatty acids (SCFA) in anionic form limits their diffusion across the absorptive membrane. The present study sought to establish the mechanism of SCFA absorption in the ostrich. 2. Epithelial tissues were taken from the sacculated part of the colon and mounted in Ussing chambers in a bathing solution. The tissues were voltage-clamped and allowed to equilibrate to obtain a baseline short circuit current (I sc). 3. Propionate (23 mm) on the mucosal side increased the I sc. The SCFA-induced I sc was completely inhibited by anoxia, ouabain (1 to 2 mm), acetazolamide (0·5 mm) and 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (0·1 mm) on the mucosal side. 4. These findings indicate that SCFA stimulate hydrogen ion secretion through an electrogenic H+-K+-ATPase, the source of hydrogen ions being carbonic anhydrase catalysed hydration of CO2. 5. Simultaneous activation of <artwork name> exchange prevents intracellular accumulation of bicarbonate ions. This system may provide hydrogen ions for protonation of SCFA anions and subsequent absorption by non-ionic diffusion.

Effect of angiotensin II on ion transport across human Fallopian tube epithelial cells in vitro.

Angiotensin II type 1 receptors have been identified in Fallopian tube epithelia. Polarized confluent human Fallopian tube epithelial cell cultures were used under short-circuit conditions to study the actions of angiotensin II on electrogenic ion transport. The results demonstrate that angiotensin II increases baseline short-circuit current, implying a net transport of negatively charged ions from a basal to apical direction. This effect was inhibited by the selective angiotensin II type 1 receptor antagonist losartan. The effects of angiotensin II on short-circuit current were rapid in onset, brief in duration, and although less than those achieved with ATP, similar in amplitude to those described for other epithelia with angiotensin II. These findings reflect a significant retention of function for these cells in monolayer culture. Immunohistochemistry using the antibody 6313/G2, which is directed against a specific sequence in the extracellular domain of the angiotensin II type 1 receptor, confirmed that the receptor was retained in cultured cells. The results indicate that angiotensin II plays a role in regulating the composition of Fallopian tube secretions.

Effect of angiotensin II on ion transport across human Fallopian tube epithelial cells in vitro

Angiotensin II type 1 receptors have been identified in Fallopian tube epithelia. Polarized confluent human Fallopian tube epithelial cell cultures were used under short-circuit conditions to study the actions of angiotensin II on electrogenic ion transport. The results demonstrate that angiotensin II increases baseline short-circuit current, implying a net transport of negatively charged ions from a basal to apical direction. This effect was inhibited by the selective angiotensin II type 1 receptor antagonist losartan. The effects of angiotensin II on short-circuit current were rapid in onset, brief in duration, and although less than those achieved with ATP, similar in amplitude to those described for other epithelia with angiotensin II. These findings reflect a significant retention of function for these cells in monolayer culture. Immunohistochemistry using the antibody 6313/G2, which is directed against a specific sequence in the extracellular domain of the angiotensin II type 1 receptor, confirmed that the receptor was retained in cultured cells. The results indicate that angiotensin II plays a role in regulating the composition of Fallopian tube secretions.

In vitro anion transport alterations and apoptosis induced by phenylbutazone in the right dorsal colon of ponies.

To study the functional and structural responses of the right dorsal colon (RDC) of ponies to phenylbutazone (PBZ) in vitro at a concentration that could be achieved in vivo. 8 adult ponies. Short circuit current and conductance were measured in mucosa from the RDC. Tissues incubated with and without HCO3- were exposed to PBZ, bumetanide, or indomethacin. Bidirectional Cl- fluxes were determined. After a baseline flux period, prostaglandin E2 (PGE2) was added to the serosal surfaces and a second flux period followed. Light and transmission electron microscopy were performed. Baseline short circuit current was diminished significantly by PBZ and indomethacin, and increased significantly after addictions of PGE2. After PGE2 was added, Cl- secretion increased significantly in tissues in HCO3- -free solutions and solutions with anti-inflammatory drugs, compared with corresponding baseline measurements and with control tissues exposed to PGE2. Bumetanide did not affect baseline short circuit current and Cl- fluxes. The predominant histologic change was apoptosis of surface epithelial cells treated with PBZ and to a lesser extent in those treated with indomethacin. Prostaglandin-induced Cl- secretion appeared to involve a transporter that might also secrete HCO3-. Both PBZ and indomethacin altered ion transport in RDC and caused apoptosis; PBZ can damage mucosa through a mechanism that could be important in vivo. The clinically harmful effect of PBZ on equine RDC in vivo could be mediated through its effects on Cl- and HCO3- secretion.

Active ion secretion and permeability of rabbit maxillary sinus epithelium, impact of staphylococcal enterotoxin B

The aim of this study was to investigate the impact of staphylococcal enterotoxin B (SEB), a model superantigen, on the physiologic functions of rabbit maxillary sinus epithelium. Rabbit sinus mucosae were separated under a surgical microscope and mounted in Ussing chambers to record short-circuit current, conductance, and permeability to horseradish peroxidase. The results showed that SEB evoked increases in sinus epithelial cell baseline short-circuit current, conductance, and permeability to horseradish peroxidase. When tumor necrosis factor-α (TNF-α) was added to the Ussing chambers, we got results similar to those obtained by SEB stimulation in vitro; the effects of SEB on sinus epithelial cells could be blocked by pretreatment with anti-TNF-α antibody. These results demonstrate that SEB is able to alter the function of sinus epithelial cells and to affect the capability of the epithelial defensive barrier, which may be mediated by TNF-α. (Otolaryngol Head Neck Surg 2000;123:120-3.)

Morphology and Epithelial Ion Transport of the Alkaline Gland in the Atlantic Stingray (Dasyatis sabina).

The alkaline glands are two fluid-filled sacs that lie on the ventral, posterior surface of each kidney in skates and rays. In this study, the morphology, transepithelial ion transport, fluid constituents, and histochemistry of the alkaline glands of the Atlantic stingray, Dasyatis sabina, were investigated. The duct from each gland joined the corresponding vas deferens and the resulting two common ducts emptied into the cloaca. Dark burgundy, aqueous fluid (pH 8.0-8.2) was secreted into the sacs by a simple columnar epithelium with extensive rough endoplasmic reticulum and large secondary lysosomes containing lipofuscin and membrane fragments. Zonulae occludentes were deep (~22 fibrils), reflecting an electrically tight epithelium (732 ohms/cm2). Carbonic anhydrase activity was localized histochemically within the intercellular spaces and less intensely in the mid-basal cytoplasm. In vitro electrophysiology showed that baseline shortcircuit current (Isc, 29.1 {mu}A/cm2) was reduced 67.0% after Cl- removal from the medium. Cl- removal also completely abolished luminal alkalinization (baseline 4.5 +/- 0.7 {mu}Eq of acid/cm2/h). Luminal exposure to the chloridebicarbonate exchange inhibitor, DIDS, reduced Isc by 38%. Simultaneous administration of DIDS and bumetanide (Na+/K+/Cl- cotransport inhibitor) to the serosal side of the tissue caused the Isc to decrease >100%. Serosal exposure to ouabain (Na-K, ATPase inhibitor) decreased Isc 48%, whereas amiloride (sodium ion channel blocker) and acetazolamide (carbonic anhydrase inhibitor) had no statistically significant effect on Isc or alkalinization rates. Taken together the results suggest the presence of apical epithelial bicarbonate exchangers that are chloride or sodium dependent, basal sodium and HCO3- transport, and an Isc that is not totally dependent on Na+-K+ ATPase.

Anti sense DNA down-regulates proteins kinase C-epsilon and enhances vasopressin-stimulated Na+ absorption in rabbit cortical collecting duct.

Hormonal activation of protein kinase C (PKC) is a major signaling mechanism regulating salt and water transport in the distal nephron. We used antisense DNA to down-regulate a PKC isoform in the rabbit cortical collecting duct (CCD) and examined its role in mediating arginine vasopressin's (AVP) effect on salt transport in the CCD. Immunoblots demonstrate that PKC-epsilon (diacylglycerol sensitive) and PKC-zeta (diacylglycerol insensitive) are the major PKC isoforms in both freshly isolated and primary cultures of rabbit CCDs. Rabbit CCDs grown on semi-permeable supports, displayed a positive baseline short circuit current (Isc), which was abolished by amiloride, demonstrating active Na+ absorption. Both AVP and 8-chloro-phenylthio-cAMP (8CPTcAMP) transiently increased Isc, however, within 40 min Isc fell below baseline. Down-regulation of PKC-epsilon, as confirmed by immunoblot, was achieved either by treatment with a PKC-epsilon-specific antisense oligonucleotide or 48 h of 1 microM PMA. In PKC-epsilon down-regulated cells, 8CPTcAMP produced a sustained, rather than transient, increase in Isc. We suggest cAMP stimulates Na+ transport, but secondary activation of PKC-epsilon results in the sustained inhibition of Na+ transport seen in response to vasopressin in the CCD.

The effect of non-steroidal anti-inflammatory drugs on the electrical properties of cultured dog tracheal epithelial cells

The effect of non-steroidal anti-inflammatory drugs (NSAIDs) on the electrical properties of primary cultures of dog tracheal epithelium has been studied. The cells used were grown with an air interface in a serum-free medium on membranes coated with human placental collagen. When mounted in Ussing chambers at 37°C, mean values for the baseline short circuit current ( I sc) and the transepithelial resistance of 65 tissue specimens from 18 dogs were 24.0 ± 3.2 μ A/cm 2 and 458 ± 128 Ω · cm 2 , respectively. These tissues had been pretreated with amiloride to abolish active Na + absorption. Under these conditions, the I sc value serves as a measure of active Cl − secretion. The results of this study revealed that the I sc across a cultured monolayer of trachea was attenuated by the tested NSAIDs, indomethacin, fulfenamic acid, mefenamic acid, aspirin, and acetaminophen, with K i's that ranged from 6.0 × 10 −5 to 2.51 × 10 −3 M. Salicylic acid had no effect on baselin I sc. Cl − channel inhibitors tested was: fulfenamic acid ⪢ indomethacin > mefenamic acid ⪢ aspirin > acetaminophen > salicylic acid. The NSAIDs also significantly inhibited both the transient, Ca 2+-dependent and the sustained, cAMP-dependent increases in I sc elicited by isoproterenol. Thus, the tested NSAIDs appeared to have an effect on the electrical properties of the cells. A similar effect of NSAIDs on ion transport across the human airway epithelium may help to reduce airway fluid secretion.

Rabbit esophageal cells possess K+ channels: Effect of hyposmotic stress on channel activity

Background: In many cell types, basolateral K+ channels are important in maintaining transepithelial Na+ absorption and regulatory volume decrease (RVD) after hyposmolar stress. However, in the esophagus the effect of K+ transport in maintaining baseline short-circuit current (SCC) (Na+ absorption) and RVD is unknown. Methods: Ussing chambers were used to evaluate changes in SCC of rabbit esophageal mucosa in response to serosal Ba2+ (4 mmol/L), quinine (1 mmol/L), and increasing serosal [K+]. To determine whether K+ channel(s) are activated in RVD, changes in SCC in response to serosal hyposmolarity (156 mOsm) were assessed in the presence or absence of serosal quinine. Results: Serosal Ba2+, quinine, or increased serosal [K+] caused a decline in baseline SCC. Serosal hyposmolarity caused an increase in SCC that was not blocked by mucosal application of amiloride (10−4 mmol/L). In contrast, serosal quinine completely blocked the hyposmolar-induced increase in SCC. Conclusions: These studies suggest that rabbit esophageal cells possess Ba2+- and quinine-sensitive basolateral K+ channel(s) that are active under baseline conditions. Potassium conductance(s) also appear to be activated by external serosal hyposmolarity and may be involved in the process of RVD.

Inhibition of neuronally mediated secretion in rat colonic mucosa by prostaglandin D2

The effect of prostaglandin D2 (PGD2) on ion transport across the mucosa of the descending colon was studied in rats. PGD2 dose-dependently decreased baseline short-circuit current of mucosa-submucosal preparations mounted either in the Ussing chamber or mounted as an everted sac. However, with the everted sac technique, the tissue was about 1000 times more sensitive to PGD2. Concomitant with the decrease in short-circuit current, PGD2 increased the mucosal-to-serosal fluxes of sodium and chloride and decreased the serosal-tomucosal flux of chloride. PGD2 inhibited the secretory action of the PGI2 analogue iloprost, PGD2α, and neurotensin. The action of these secretagogues was dependent on the presence of the submucosal plexus. In contrast, PGD2 had no effect on the increase in short-circuit current caused by PGD2, substance P, or serotonin, the actions of which were not dependent on the presence of the submucosal plexus. The results indicate that the action site of the antisecretory mechanism of PGD2 is localized in the secretomotor neurons.

Steroid hormones: modulators of Na+ absorption and Cl- secretion in cultured tracheal epithelia.

Cultures of tracheal epithelial cells from adult nonpregnant female rabbits maintained a different balance of Na+ absorption and Cl- secretion from cells of pregnant animals and fetal cultures. Cells of nonpregnant rabbits had a large Na+ absorptive current (11 microA/cm2 or 53% of baseline short circuit current) and a small Cl- secretory current (2.4 microA/cm2 or 11%). In contrast, fetal cells had greater Cl- secretion (15.1 microA/cm2 or 61%) and much less Na+ absorption (1.6 microA/cm2 or less than 10%). Cells of pregnant animals had amounts of Cl- secretion (11.4 microA/cm2 or 41%) and Na+ absorption (6.7 microA/cm2 or 22%) that were in between that of fetal and adult nonpregnant animals. In addition to these striking differences in baseline ion transport among the three groups, epinephrine was found to stimulate Cl- secretion in cells from pregnant rabbits, both Na+ absorption and Cl- secretion in cells from nonpregnant animals, and neither in fetal cells. Steroid hormones of pregnancy, progesterone and estradiol, when added to cells cultured from nonpregnant rabbits, altered the magnitudes of the individual transport pathways to mimic cells cultured from pregnant animals. In addition to changing baseline transport, these steroids modified the epinephrine-stimulated currents to resemble those in the fetus and in pregnancy. We conclude that steroid hormones regulate the balance of Na+ absorption and Cl- secretion in tracheal epithelia in utero and in adulthood.

The involvement of afferent nerve terminals in the stimulation of ion transport by bradykinin in rat isolated colon.

1. The actions of bradykinin (Bk) were investigated on rat colon epithelium preparations that had been stripped of the muscle layers. The electrogenic ion flux was monitored by measuring changes in the short circuit current (SCC) produced by addition of drugs. Bk, administered to the basolateral side, but not apical side, of the epithelium evoked an increase in SCC which was separable into two distinct components, both of which were mediated mainly by chloride efflux. 2. The early component was robust, reproducible and exhibited clear concentration-dependency with an EC50 of 6.2 nM. The second phase of the response exhibited a much slower time course than the first phase and diminished amplitude with repeated applications of Bk. 3. In preparations of unstripped epithelium, bradykinin (Bk) evoked mainly a slow neurogenic response which was attenuated or abolished by tetrodotoxin (TTX). When the epithelium was stripped off, TTX had little effect either on the baseline SCC or on responses to Bk. 4. Perfusion with zero calcium solution did not affect the early phase but abolished the late phase of the Bk response. Verapamil (20 microM), but not nifedipine (20 microM), also attenuated the later phase of the response. 5. Capsaicin (2 microM) administered to the basolateral, but not the apical, side produced an increase in SCC. Following desensitization to capsaicin the second phase of the response to Bk was abolished with little effect on the initial response to Bk. 6. The data suggest that Bk increases the efflux of chloride ions across the colonic epithelium in at least two ways: (a) by an action on the epithelial cells and (b) by an action on neuronal elements within the epithelium. This latter effect of Bk is due to stimulation of capsaicin-sensitive nerve terminals within the mucosa of the colon epithelium causing the release of a mediator which is responsible for the second phase of the response to Bk.

Muscarinic receptor subtypes mediating the mucosal response to neural stimulation of guinea pig ileum

Muscarinic receptors involved in the secretory response evoked by electrical stimulation of submucosal neurons were investigated in muscle-stripped flat sheets of guinea pig ileum set up in flux chambers. Neural stimulation produced a biphasic increase in short-circuit current due to active chloride secretion. Atropine and 4-diphenylacetoxy-N-methylpiperadine methiodide (4-DAMP) (10(-7) M) were more potent inhibitors of the cholinergic phase of the response than was pirenzepine. Dose-dependent increases in base-line short-circuit current were evoked by carbachol and bethanechol; 4-hydroxy-2-butynyl trimethylammonium chloride (McN A343) produced a much smaller effect. Tetrodotoxin abolished the effects of McN A343 but did not alter the responses of carbachol and bethanechol. McN A343 significantly reduced the cholinergic phase of the neurally evoked response and caused a rightward shift of the carbachol dose-response curve. All muscarinic compounds inhibited [3H]quinuclidinyl benzilate binding to membranes from mucosal scrapings, with a rank order of potency of 4-DAMP greater than pirenzepine greater than McN A343 greater than carbachol greater than bethanechol. These results suggest that acetylcholine released from submucosal neurons mediates chloride secretion by interacting with muscarinic cholinergic receptors that display a high binding affinity for 4-DAMP. Activation of neural muscarinic receptors makes a relatively small contribution to the overall secretory response.

Vasoactive intestinal polypeptide actions on the guinea pig intestinal mucosa during neural stimulation

Electrical stimulation of the mucosal innervation of the guinea pig ileum results in an increase in chloride secretion that is mediated in part by excitation of muscarinic cholinergic receptors on enterocytes. This study investigated the involvement of vasoactive intestinal peptide in the cholinergic and noncholinergic phases of the secretory response evoked by electrical stimulation of submucosal neurons in the guinea pig ileum. Flat sheets of ileum set up in Ussing flux chambers responded to exogenous vasoactive intestinal peptide by an increase in baseline short-circuit current which was reduced by furosemide and by vasoactive intestinal peptide antiserum. When submucosal neurons were electrically stimulated, a biphasic change in short-circuit current was evoked. Vasoactive intestinal peptide, forskolin, and isobutylmethylxanthine enhanced the cholinergic portion of the response, whereas the antiserum prevented or reduced the effects of the peptide but not of forskolin. In the presence of atropine to eliminate the cholinergically mediated response, vasoactive intestinal peptide reduced the noncholinergic phase of the response and its action was prevented by the antiserum. Vasoactive intestinal peptide enhanced the increase in short-circuit current evoked by the muscarinic agonist bethanechol. These results demonstrate that vasoactive intestinal peptide and other substances that stimulate secretion by increasing cyclic 3′,5′-adenosine monophosphate levels in enterocytes potentiate the calcium-dependent, cholinergic phase of the chloride secretory response evoked by neural stimulation of the guinea pig ileum. No evidence was found for vasoactive intestinal peptide as the mediator of the noncholinergic phase of the response.

Ion transport in human cecum, transverse colon, and sigmoid colon in vitro: Baseline and response to electrical stimulation of intrinsic nerves

In a flux chamber study of ion transport in human colon, we compared baseline rates with those measured during electrical stimulation of intrinsic nerves. In baseline studies, sodium was absorbed throughout, but maximally in transverse colon. In cecum, sodium absorption accounted for the short circuit current and chloride was not absorbed. Chloride was absorbed in transverse and sigmoid colon, however. Residual current was minimal in cecum and transverse colon, but increased in sigmoid colon. Neural stimulation caused chloride secretion in cecum, reduced chloride absorption in sigmoid colon, but caused no change in transverse colon; sodium absorption decreased in cecum. A neurotransmitter of unknown identity affects baseline short circuit current in sigmoid colon. Half of the increase in short circuit caused by neural stimulation in sigmoid colon is mediated by muscarinic receptors. The identity of the other transmitter(s) is not known. It is not substance P or histamine. The three divisions of the colon differ in relative rates of baseline ion transport and in their transport responses to intrinsic nerve stimulation.