Biomedical subjects

K M Sanders

Publications and source records attributed to K M Sanders.

At least 109 records · Page 6Linked to original sources

PubMed1997

Effects of a novel guanylate cyclase inhibitor on nitric oxide-dependent inhibitory neurotransmission in canine proximal colon.

1. Previous studies suggested that nitric oxide (NO) may cause hyperpolarization and relaxation of canine colonic smooth muscle by both cGMP-dependent and cGMP-independent mechanisms. This hypothesis was tested using 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ), a novel inhibitor of NO-stimulated guanylate cyclase. 2. In the presence of histamine (30 microM), atropine and indomethacin (both at 1 microM), electrical field stimulation of intrinsic neurons (EFS; 5 Hz) produced inhibition of phasic contractile activity that is due to NO synthesis. ODQ caused a concentration-dependent block of this response (10 nM to 10 microM). 3. Inhibitory junction potentials (IJPs) due to NO synthesis were recorded from muscle cells located near the myenteric border of the circular muscle layer, using intracellular microelectrodes. IJPs were abolished by ODQ (1-10 microM). 4. EFS (10-20 Hz) produced frequency-dependent inhibition of electrical slow waves recorded from cells located near the submucosal surface of the circular muscle layer. This inhibition is due to NO synthesis, and it was abolished by ODQ (1-10 microM). 5. Hyperpolarization and relaxation produced by an NO donor, sodium nitroprusside, were abolished by ODQ pretreatment (1-10 microM). In contrast, inhibitory responses to 8-Br-cGMP (1 mM) were unaffected by ODQ. 6. ODQ alone (1-10 microM) had no significant effect on spontaneous electrical or phasic contractile activity. In tissues pre-treated with L-NAME (300 microM), ODQ decreased the amplitude of spontaneous or histamine-stimulated phasic contractile activity. 7. These results suggest that electrical and mechanical effects of endogenously released and exogenously applied NO in canine colon are largely due to cGMP synthesis by ODQ-sensitive soluble guanylate cyclase. No evidence to support a direct (cGMP-independent) mechanism of NO action was found. ODQ also appears to cause a non-specific inhibition of muscle contractile activity; however, this effect does not contribute to block of NO-dependent effects.

Animals↗

PubMed1997

Excitatory and inhibitory actions of vasopressin on colonic excitation-contraction coupling in dogs.

BACKGROUND & AIMS: Vasopressin, a circulating hormone and agent used to treat gastrointestinal bleeding, affects gastrointestinal motility. Both excitatory and inhibitory responses that may be caused by species differences or concentration-dependent effects of this hormone have been reported. This study examined the actions of vasopressin on the canine colon and studied the mechanism underlying the excitatory effects of vasopressin. METHODS: Intracellular microelectrodes and isometric force measurements were used to measure vasopressin responses in muscle strips. The patch-clamp technique was used to record arginine vasopressin activation of nonselective cation currents in isolated myocytes. RESULTS: Vasopressin (10(-12) to 10(-9) mol/L) increased electrical slow waves and enhanced phasic contractions. Atropine and tetrodotoxin did not alter the responses to vasopressin. In myocytes, vasopressin activated a nonselective cation conductance at concentrations that enhanced slow waves. Vasopressin (> 5 x 10(-8) mol/L) caused a tetrodotoxin- or NG-nitro-L-arginine methyl ester-insensitive inhibition of slow waves and contractile activity. A V1-receptor antagonist shifted the effects of vasopressin to higher concentrations. Immunohistochemistry showed expression of vasopressin-like immunoreactivity within the colonic wall, suggesting that local concentrations may exceed circulating levels. CONCLUSIONS: These data document the concentration dependence of the biphasic effects of vasopressin and provide a mechanism for the excitatory effects of vasopressin at physiological levels.

Acetylcholine↗

PubMed1997

The soluble interleukin-2 receptor predicts mortality in older hospitalized men.

BACKGROUND: There is an inverse relationship between the soluble interleukin-2 receptor (sIL-2R) and serum albumin, cholesterol, transferrin, prealbumin, and hemoglobin. Inasmuch as low serum albumin and cholesterol have been associated with excess mortality, we hypothesized that elevated sIL-2R would predict mortality in older adults. OBJECTIVE: To determine if elevated sIL-2R predicts mortality in patients on a geriatric rehabilitation unit. DESIGN: Prospective cohort. SETTING: University-affiliated VA medical center. PARTICIPANTS: Seventy-two male patients aged greater than 60 years admitted to a geriatric rehabilitation unit. Patients with severe hepatic or renal disease were excluded. MEASUREMENTS: We measured serum albumin, prealbumin, cholesterol, transferrin, hemoglobin, body mass index (BMI), C-reactive protein (CRP), and sIL-2R upon admission. Subjects were followed for 1 year. RESULTS: Low serum albumin, prealbumin, and hemoglobin and high sIL-2R and CRP predicted 1-year mortality on univariate analysis. When these predictors were included as covariates in a Cox regression model, only sIL-2R was a significant independent predictor of mortality (P = .043). Multiple linear regression with the above covariates revealed that only sIL-2R predicted time to death at (P = .003). CONCLUSIONS: High sIL-2R and CRP and low albumin, prealbumin, and hemoglobin predicted mortality using univariate analysis on a rehabilitation unit. However, with multivariate analysis, sIL-2R was the sole predictor of mortality.

Aged↗

PubMed1997

Small-conductance Ca(2+)-dependent K+ channels activated by ATP in murine colonic smooth muscle.

The patch-clamp technique was used to determine the ionic conductances activated by ATP in murine colonic smooth muscle cells. Extracellular ATP, UTP, and 2-methylthioadenosine 5'-triphosphate (2-MeS-ATP) increased outward currents in cells with amphotericin B-perforated patches. ATP (0.5-1 mM) did not affect whole cell currents of cells dialyzed with solutions containing ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. Apamin (3 x 10(-7) M) reduced the outward current activated by ATP by 32 +/- 5%. Single channel recordings from cell-attached patches showed that ATP, UTP, and 2-MeS-ATP increased the open probability of small-conductance, Ca(2+)-dependent K+ channels with a slope conductance of 5.3 +/- 0.02 pS. Caffeine (500 microM) enhanced the open probability of the small-conductance K+ channels, and ATP had no effect after caffeine. Pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid tetrasodium (PPADS, 10(-4) M), a nonselective P2 receptor antagonist, prevented the increase in open probability caused by ATP and 2-MeS-ATP. PPADS had no effect on the response to caffeine. ATP-induced hyperpolarization in the murine colon may be mediated by P2y-induced release of Ca2+ from intracellular stores and activation of the 5.3-pS Ca(2+)-activated K+ channels.

Adenosine Triphosphate↗

PubMed1997

Maintenance of morphology and function of canine proximal colon smooth muscle in organ culture.

We have determined that serum source plays a critical role in optimizing conditions for an organ culture model of canine proximal colon. Previous studies using equine serum in the medium have shown that some properties of canine colonic smooth muscle can be maintained in organ culture. However, many characteristics of the tissue were altered by the culture conditions. The aims of the present study were to determine whether serum isolated from canine blood would improve the preservation of physiological properties of canine proximal colon in organ culture. Strips of canine colonic smooth muscle were cultured in 10% canine serum medium, and electrical, mechanical, morphological, and molecular analyses were performed after 0, 3, and 6 days in culture. Unlike organ culture in equine serum, in which Na+-K+-adenosinetriphosphatase (Na+-K+-ATPase) expression declined, culture in canine serum maintained Na+-K+-ATPase expression, and resting membrane potential of smooth muscle cells along the submucosal surface of the circular muscle in cultured tissue remained unchanged during the culture period. Increased sensitivity in the contractile response to acetylcholine, previously observed with tissues cultured in equine serum, was not observed. However, the mechanical performance of the muscle (maximal contractile activity) declined over time in culture. Ultrastructural organization of cellular organelles and myofilaments remained intact in the majority of cells; however, some cells possessed regions devoid of contractile filaments. The results of these studies suggest that organ cultured strips of smooth muscle may provide a useful tool for evaluating electrical and mechanical events in conjunction with molecular analysis of functional components.

Animals↗

PubMed1996

Enteric neurons express Steel factor-lacZ transgene in the murine gastrointestinal tract.

The cells that express Steel factor (SLF) in the gastrointestinal (GI) tract were studied using SLF-lacZ transgenic mice. Expression, detected by beta-galactosidase histochemistry, was evident in cells between the circular and longitudinal muscle layers in the GI tract. Double staining with antibodies specific for the neural markers, PGP 9.5, MAP2 and c-Ret, showed that SLF-lacZ positive cells were enteric neurons. Enteroglia did not express SLF-lacZ. The distribution of expressing cells was complimentary to the expression of c-Kit in myenteric interstitial cells.

Animals↗

PubMed1996

Interstitial cells of Cajal mediate inhibitory neurotransmission in the stomach.

The structural relationships between interstitial cells of Cajal (ICC), varicose nerve fibers, and smooth muscle cells in the gastrointestinal tract have led to the suggestion that ICC may be involved in or mediate enteric neurotransmission. We characterized the distribution of ICC in the murine stomach and found two distinct classes on the basis of morphology and immunoreactivity to antibodies against c-Kit receptors. ICC with multiple processes formed a network in the myenteric plexus region from corpus to pylorus. Spindle-shaped ICC were found within the circular and longitudinal muscle layers (IC-IM) throughout the stomach. The density of these cells was greatest in the proximal stomach. IC-IM ran along nerve fibers and were closely associated with nerve terminals and adjacent smooth muscle cells. IC-IM failed to develop in mice with mutations in c-kit. Therefore, we used W/W(V) mutants to test whether IC-IM mediate neural inputs in muscles of the gastric fundus. The distribution of inhibitory nerves in the stomachs of c-kit mutants was normal, but NO-dependent inhibitory neuro-regulation was greatly reduced. Smooth muscle tissues of W/W(V) mutants relaxed in response to exogenous sodium nitroprusside, but the membrane potential effects of sodium nitroprusside were attenuated. These data suggest that IC-IM play a critical serial role in NO-dependent neurotransmission: the cellular mechanism(s) responsible for transducing NO into electrical responses may be expressed in IC-IM. Loss of these cells causes loss of electrical responsiveness and greatly reduces responses to nitrergic nerve stimulation.

Animals↗

PubMed1996

Activation of delayed rectifier potassium channels in canine proximal colon by vasoactive intestinal peptide.

1. Vasoactive intestinal peptide (VIP) inhibits phasic contractions and tone of gastrointestinal smooth muscles. This study examines electrical mechanisms that may mediate the inhibitory actions of VIP. 2. Electrical slow waves were recorded from canine proximal colon circular muscles. VIP (0.1 microM) decreased basal slow wave frequency but had no effect on amplitude or duration. When slow waves were enhanced with Bay K 8644 (1 microM), VIP decreased slow wave duration and inhibited contractions. 3. VIP inhibited slow waves and phasic contractions stimulated by tetraethylammonium chloride (TEA; 10 mM), but did not significantly reduce events stimulated by 4-amino-pyridine (4-AP; 10 mM). 4. Whole-cell outward currents were recorded from isolated myocytes, using the amphotericin B perforated patch technique. VIP (1 microM) increased charybdotoxin-insensitive outward currents. 5. Single voltage-dependent K+ channels were recorded in cell-attached patches. VIP increased reversibly the open probability, mean open time and mean burst duration of 4-AP-sensitive, charybdotoxin-insensitive K+ channels (KDR1). Two additional 4-AP- and charybdotoxin-insensitive K+ channels (approximately 90 pS and < 4 pS) were also observed in these patches, but were not significantly affected by VIP. 6. In summary, the effects of VIP on electrical slow waves may be due, in part, to activation of 4-AP-sensitive, 'delayed rectifier' K+ channels. Activation of these channels may contribute to premature slow wave repolarization, reduced Ca2+ entry, and inhibition of contractile force.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1996

Expression of nitric oxide synthase in mucosal cells of the canine colon.

The expression of nitric oxide synthase (NOS) in the mucosa of the canine colon was investigated with in situ hybridzation, immunohistochemistry (using isoform specific antibodies), western analysis, and NADPH diaphorase (NADPH-d) histochemistry. In situ hybridization using a common probe for known isoforms of NOS showed that NOS mRNA was strongly expressed in mucosal cells. A gradient in the degree of hybridization was noted from the base of the crypts to the luminal surface. This gradient was also apparent using an endothelial NOS (eNOS)-specific probe. Neural NOS-like immunoreactivity (nNOS-LI) was observed in columnar epithelial cells, and the same population of cells was stained with NADPH-d. Endothelial NOS-like immunoreactivity (eNOS-LI) was also found in mucosal cells; however, this eNOS-LI was confined to mucous cells. These cells were not stained with NADPH-d. The existence of eNOS in mucosal cells was confirmed by in situ hybridization using the probe which specifically hybridized with mRNA of eNOS and by western blots which demonstrated the expression of a 135-kDa protein in mucosal homogenates. The differential expression of NOS isoforms and the gradient in expression along the length of the crypts suggest complex roles for NO in the development of colonic epithelial cells and in secretion and transport functions of the colonic mucosa.

Animals↗

PubMed1996

Regulation of smooth muscle delayed rectifier K+ channels by protein kinase A.

We identified voltage-activated K+ channels in freshly dispersed smooth muscle cells from the circular layer of the canine colon in patch-clamp experiments using 200 nM charybdotoxin to suppress 270-pS Ca2+-activated K+ channels (BK channels). Three channel types were distinguished in symmetrical 140 mM KCl solutions: 19.5 +/- 1.7 pS channels (KDR1), 90.6 +/- 5.4 pS channels (KDR2) and 149 +/- 4 pS intermediate-conductance Ca2+-activated K+ channels (IK channels). All three types showed an increase in open probability with membrane depolarization. Ensemble average current from KDR1 channels inactivated with a time constant of 1.7 +/- 0.1 s at +60 mV test potential, while KDR2 and IK channels did not show inactivation. IK channels were activated by free cytoplasmic [Ca2+] (10(-6 )M) but were insensitive to 4-aminopyridine (4-AP, 10 mM) and intracellular tetraethylammonium (TEA, 1 mM). KDR1 channels were sensitive to 4-AP (10 mM) and intracellular TEA (1-10 mM) but not to Ca2+. KDR2 channels did not have a consistent pharmacological profile, suggesting that this class may be comprised of several subtypes. At +40 mV membrane potential, the catalytic subunit of protein kinase A (PKA) increased the open probability of KDR1 channels 3.4-fold and of KDR2 channels 3.9-fold, but had no effect on IK channels. In the absence of Mg-ATP, PKA did not affect channel open probabilities. At physiological membrane potentials (-60 mV) only openings of KDR1 channels could be induced by PKA, suggesting that these 4-AP-sensitive 20-pS K+ channels are primarily responsible for the cAMP-mediated hyperpolarization of colonic smooth muscle cells.

4-Aminopyridine↗

PubMed1996

A case for interstitial cells of Cajal as pacemakers and mediators of neurotransmission in the gastrointestinal tract.

Electrical rhythmicity in gastrointestinal muscles has been studied for a century, but the pacemakers driving this phenomenon have been elusive. Anatomic studies suggest that interstitial cells of Cajal (ICC) may be pacemakers and conductors of electrical activity. ICC may also mediate neurotransmission from enteric neurons. Functional evaluations of ICC include the following. (1) Electrophysiology experiments on dissected muscle strips show that slow waves originate from specific sites. These pacemaker areas are populated by networks of ICC that make gap junctions with smooth muscle cells. Removal of pacemaker regions interferes with slow wave generation and propagation. (2) Chemicals that label ICC histochemically can damage ICC and abolish rhythmicity. (3) isolated ICC are spontaneously active, and several voltage-dependent ion channels, including a low-threshold Ca2+ conductance, are expressed. (4) ICC are innervated by enteric neurons, and they respond to neurotransmitters. ICC may produce nitric oxide and amplify inhibitory neurotransmission. (5) Some classes of ICC fall to develop in animals with mutations in c-kit or stem cell factor, the ligand for c-Kit receptors. Without ICC, electrical slow waves are absent. Many questions remain about the function of ICC, but modern technologies should now facilitate rapid progress toward determining the role of these cells in normal physiology and pathological conditions.

Animals↗

PubMed1996

Hyperpolarization and inhibition of contraction mediated by nitric oxide released from enteric inhibitory neurones in guinea-pig taenia coli.

1. Inhibition of nitric oxide synthase by NG-nitro-L-arginine (L-NNA) reduced the neurogenic relaxation of precontracted taenia coli only in the absence of atropine. The membrane hyperpolarization associated with the neurogenic relaxation was also reduced by inhibition of NOS only when atropine was absent. 2. The membrane hyperpolarization associated with the neurogenic relaxation of the taenia coli was inhibited by oxyhaemoglobin only in the absence of atropine. In the presence of atropine, oxyhaemoglobin did not reduce the i.j.p. or nerve evoked relaxation. 3. Inhibition of NOS by L-NNA did not affect the overflow of [3H]-ACh in response to electrical field stimulation (EFS), suggesting that, under the conditions of our experiments, endogenous NO did not modulate release of ACh. Sodium nitroprusside also had no effect on the neurogenic overflow of [3H]-ACh; however, noradrenaline significantly reduced [3H]-ACh overflow. 4. In summary, the postjunctional effects of neurally-released NO are not apparent in guinea-pig taenia coli when atropine is present. This implies muscarinic regulation of NO release or muscarinic regulation of another excitatory substance, such as tachykinin(s), that, when blocked, masks the postjunctional effects of NO. These data, together with previous studies, suggest a possible regulatory role for NO in enteric neurotransmission that may be more prominent in some species or tissues than others.

Acetylcholine↗

PubMed1996

Wortmannin inhibits contraction without altering electrical activity in canine gastric smooth muscle.

Wortmannin, an inhibitor of myosin light-chain kinase (10-30 microM), completely and irreversibly abolished (in 75% of tissues from canine gastric antrum) phase contractions caused by slow waves with no significant effects on resting membrane potential or the frequency, amplitude, or duration of spontaneous slow waves. Responses to agents that normally cause hyperpolarization (cromakalim, sodium nitroprusside, and forskolin) were unaffected by wortmannin treatment. It was also possible to study the excitatory effects of agents and conditions that normally result in loss of intracellular impalements: 1) elevated extracellular K+ concentrations altered membrane potential close to values predicted by the Nernst equation, and 2) high concentrations of acetylcholine produced depolarization and rapid oscillations in membrane potential coincident with contractile activity. Cholinergic increases in myosin light-chain phosphorylation and contractions were partially blocked by wortmannin. In canine antrum, wortmannin inhibition of contraction was irreversible, although in other tissue types, partial recovery of contractions was observed when wortmannin was removed. Wortmannin can be a useful agent to investigate the electrophysiology of some smooth muscles when movement might lead to recording artifacts or loss of signal.

Androstadienes↗

PubMed1996

Regulation of ion channels in smooth muscles by calcium.

Intracellular Ca2+ concentration ([Ca2+]i) plays a central role in regulating tone and contractility of smooth muscle cells. In contrast to the "classic" model of electromechanical coupling where membrane potential determines [Ca2+]i, it is now well established that [Ca2+]i in turn may also affect membrane potential by modulating open probabilities of ion channels. Activation by [Ca2+]i of large-conductance K+ channels, Cl- channels, and nonselective cation channels has been described, as well as block of delayed rectifier K+ channels by [Ca2+]i and [Ca2+]i-induced inactivation of Ca2+ channels. Therefore, a network consisting of positive- and negative-feedback loops regulates [Ca2+]i as well as membrane potential. In this context, we review the properties of Ca(2+)-dependent ion channels and their functional role in vascular and visceral smooth muscles. Any alteration of the "Ca2+ sensitivity" of ion channels is expected to have a profound effect on the reciprocal relationship between membrane potential and [Ca2+]i. Already several molecular factors determining Ca2+ sensitivity of Ca(2+)-activated K+ channels have been identified. We provide a working definition for Ca2+ sensitivity.

Animals↗

PubMed1996

Modulation of Ca2+ current in canine colonic myocytes by cyclic nucleotide-dependent mechanisms.

Regulation of Ca2+ currents by cyclic nucleotide-dependent mechanisms was studied in circular muscle cells isolated from canine proximal colon. Whole cell Ca2+ currents were recorded at 32 degrees C with the use of amphotericin B-perforated patches. The effects of several agents known to increase levels of adenosine 3',5'-cyclic monophosphate (cAMP) were tested. Vasoactive intestinal peptide (VIP) and isoproterenol (10(-7) to 10(-5) M) increased Ca2+ current in a concentration-dependent manner. Forskolin (10(-7) M) and dibutyryl cAMP (10(-6) to 10(-5) M) also increased Ca2+ current. Higher concentrations of forskolin (10(-6) to 10(-5) M) caused inhibition of Ca2+ current. Low concentrations (10(-5) to 10(-7) M) of dibutyryl cAMP or 8-bromo-cAMP caused concentration-dependent enhancement in Ca2+ current, and these effects were reversible on washout of the cAMP analogues. When the concentration of cAMP analogues was increased (10(-3) to 10(-4) M), we observed inhibition of Ca2+ current similar to the effects of forskolin. Membrane-permeable analogues of guanosine 3',5'-cyclic monophosphate produced exclusively inhibitory effects. The nonspecific protein kinase inhibitor H-7 (up to 60 microM) failed to block the effects of VIP, isoproterenol, and forskolin, and it produced inhibitory effects on Ca2+ current, independent of agonist stimulation. The data suggest that low levels of cAMP may, via phosphorylation by protein kinase A, enhance L-type Ca2+ current, but higher concentrations of cAMP may "cross over" and activate protein kinase G. Phosphorylation by protein kinase G appears to produce a dominant inhibition of Ca2+ current.

Adrenergic alpha-Agonists↗

PubMed1996

Cloning and expression of the large-conductance Ca(2+)-activated K+ channel from colonic smooth muscle.

We have cloned cDNAs encoding the alpha- and beta-subunits of a large-conductance Ca(2+)-activated K+ channel (BK channel) from canine colonic smooth muscle (cslo-alpha and cslo-beta). Nucleotide sequence homology of cslo-alpha with mslo and dslo suggests that it is the canine homologue of these genes. The carboxy-terminal end of the protein is the most diverse between species, and we have also found alternative exons in cslo-alpha in this region. We have identified a unique splice site in the carboxy-terminal region of cslo-alpha, which we term site 5. Northern analysis demonstrates expression of both alpha- and beta-subunits in all canine vascular and visceral smooth muscles tested. Expression of alpha-1 alone and alpha + beta-subunit cRNA in Xenopus oocytes results in a Ca(2+)- and voltage-dependent conductance. The activity of alpha/beta-channels, measured as either changes in the voltage of half-maximal activation (V0.5) in open probability (NP0) or in the normalized conductance (G/Cmax), was more sensitive to [Ca2+]free than channels composed of the alpha-subunit alone. Neither alpha- nor alpha/beta-channels expressed in membrane patches of Xenopus oocytes were found to be regulated by protein kinase G.

Alternative Splicing↗

PubMed1996

Failure to record delirium as a complication of intra-aortic balloon pump treatment: a retrospective study.

This study was conducted to determine whether or not diagnosis and treatment of delirium among patients treated with the intra-aortic balloon pump (IABP) correlates with the recording of this complication on discharge records. Since prior episodes of delirium are one of the few clear risk factors for future episodes of delirium, accurate recording of delirium on the discharge summary and list of discharge diagnoses is useful to clinicians. A retrospective review of the charts of all patients (N = 198) who underwent placement of an IABP during 1988; assessment of the type and frequency of medical and neuropsychiatric complications during IABP treatment; and comparison of chart review findings with the Massachusetts General Hospital's computer-generated lists of discharge diagnoses for the same IABP-treated patients was completed. Only 12% of patients diagnosed and treated for delirium had delirium recorded as a discharge diagnosis. In contrast, 44% and 52% of patients who had been diagnosed and treated for cerebrovascular accident and pneumonia, respectively, had these diagnoses recorded among the discharge diagnoses. Receiving a discharge diagnosis of organic brain syndrome increased the likelihood that delirium was recorded as a discharge diagnosis. Delirium is underdiagnosed as a complication associated with IABP-treatment and is under-reported on the list of discharge diagnoses, even when it is diagnosed. Further study is warranted to determine if making the diagnosis of delirium during a patient's hospital course and recording it is a complication at the time of discharge is translated into a higher level of preparedness by physicians during subsequent hospitalizations.

Aged↗

PubMed1996

Involvement of nitric oxide in neuromuscular transmission in canine proximal colon.

The evidence is very strong that NO serves as a neurotransmitter in some autonomic neurons. In the canine proximal colon, NOS is localized in fibers and varicosities of inhibitory motor neurons that course through the muscle layers. Excitation of inhibitory neurons enhances Ca2+ entry into varicosities and activates NOS. In the GI tract enteric inhibitory neurons not only possess the ability to synthesize NO, they may also recycle the by-product, citrulline, back to arginine, thus sustaining inhibitory neurotransmission. NO appears to diffuse freely from nerve terminals and into nearby postjunctional cells. Interstitial cells appear to be innervated by nerves that release NO, and postjunctional effects in these cells include production of cGMP and synthesis of additional NO. In smooth muscle cells NO signals are transduced by guanylate cyclase, the production of cGMP, and activation of protein kinase G, but direct stimulation by NO of some cellular effectors, such as K+ channels also appears to play a role. Responses of smooth muscle cells include activation of K+ channels, inhibition of Ca2+ channels, and a reduction in the sensitivity of the contractile apparatus to Ca2+. All of these factors may contribute to the reduction in mechanical activity produced by stimulation of enteric inhibitory neurons. NO-dependent neurotransmission is critical for many of the physiological processes of the GI tract, such as relaxation of sphincters, gastric accommodation, and receptive relaxation during feeding, and the descending inhibition arc of the peristaltic reflex.

Animals↗