Biomedical subjects

R A Bywater

Publications and source records attributed to R A Bywater.

At least 19 recordsLinked to original sources

PubMed2002

Enteric nerve stimulation evokes a premature colonic migrating motor complex in mouse.

The effects of enteric nerve stimuli were investigated on spontaneously occurring colonic migrating motor complexes (CMMCs) in isolated mouse colon. Changes in circular smooth muscle tension were recorded simultaneously from the proximal, mid and distal regions of an in vitro preparation of whole mouse colon at 36 +/- 1 degrees C. The CMMCs were recorded from all preparations with a mean interval between contractions ranging from 135.2 +/- 9.3 to 163.3 +/- 22.4 s. The CMMCs migrated spontaneously from the proximal to distal colon and were abolished by tetrodotoxin (1 micromol L-1). In approximately half of all trials (57 of 103, n = 31), trains of stimuli (20 Hz, 2-5 s, 1 ms, 40-70 V) delivered to the mid or distal regions of colon, during the intervals between CMMCs, elicited a premature CMMC. However, similar trains of stimuli delivered to the proximal colon were without similar effects (33 trials, n = 13). It is suggested that in isolated whole mouse colon, CMMCs can be evoked prematurely by trains of electrical stimuli applied to the enteric nerves. The observation that nerve stimuli failed to evoke a premature CMMC from the proximal colon suggests that selective activation of functional ascending pathways may be required to initiate a premature CMMC.

Anesthetics, Local↗

PubMed2001

Endogenous nitric oxide release modulates the direction and frequency of colonic migrating motor complexes in the isolated mouse colon.

Spontaneous colonic migrating motor complexes (CMMCs) were recorded from circular muscle at three sites along the isolated mouse colon. The interval between CMMCs was decreased from approximately 3 min in control solution, by approximately 55% in a nitric oxide synthase (NOS) inhibitor, N-nitro-L-arginine (L-NNA; 100 micromol L-1). This was associated with a shift in migration direction of CMMCs, such that CMMCs migrated in an oral direction. Application of the endogenous substrate for NOS, L-arginine, at a low concentration used to mimic plasma concentration (134 micromol L-1), or a high concentration (5 mmol L-1) suppressed CMMCs (for at least 15 min) which were replaced by high frequency (10-15 min-1), short duration (half width approximately 1.5 s) contractions of variable amplitudes (largest in the proximal region) until CMMCs resumed. CMMCs remained in the presence of D-arginine (134 micromol L-1 and 5 mmol L-1). Apamin (250 nmol L-1) did not alter the interval between CMMCs, however, additional nonmigrating contractions were observed between the CMMCs in the distal region. In addition to its effects on smooth muscle tone, NO, but not apamin-sensitive channels, plays an important role in suppressing the frequency of migrating contractions in the isolated mouse colon. Consideration should be given to the inclusion of L-arginine, in in vitro experiments where there may be spontaneous activity in NOS containing neurones.

Animals↗

PubMed2001

Ongoing nicotinic and non-nicotinic inputs to inhibitory neurons in the mouse colon.

1. Intracellular microelectrodes were used to record spontaneous and evoked inhibitory junction potentials (IJP) from the circular muscle layer of the mid-distal region of the mouse isolated colon in the presence of nifedipine (1 micromol/L) and hyoscine (1 micromol/L). 2. The length of the tissue preparation (> 1 cm) or the presence of the mucosa had no effect on the frequency of spontaneous IJP. 3. Hexamethonium (500 micromol/L) reduced the frequency of spontaneous IJP to approximately 70% of the control frequency, whereas D-tubocurarine (280 micromol/L) reduced the frequency to approximately 17% of control. Apamin (250 nmol/L) abolished all spontaneous IJP activity. 4. The greater inhibition of spontaneous IJP in the presence of D-tubocurarine compared with hexamethonium is discussed as a possible 'apamin-like' effect. 5. Although electrically evoked IJP (single pulse at 15 V, 0.6 msec) were not significantly affected by hexamethonium, D-tubocurarine and apamin reduced the amplitude of evoked IJP to approximately 65 and 50% of control, respectively. 6. These results suggest that the properties of spontaneous IJP cannot be inferred by a study of evoked IJP alone.

Animals↗

PubMed2000

Chronotropic action of 5-hydroxytryptamine (5-HT) on colonic migrating motor complexes (CMMCs) in the isolated mouse colon.

The effects of 5-hydroxytryptamine (5-HT) and related drugs on colonic migrating motor complexes (CMMCs) were evaluated in isolated colons from the heterozygotes of pie-bald lethal mice. 5-HT produced a dose-related increase in the frequency of CMMCs without any change in the amplitude or duration of the CMMC contractions themselves. The 5-HT(2) agonist, alpha-methyl 5-HT, (100 nM-1 microM) increased the frequency of CMMCs whilst the 5-HT(3) agonist, 2-methyl 5-HT, did so at 10 microM. The 5-HT(4) agonist, 5-methoxy dimethyl tryptamine oxalate did not alter the frequency of CMMCs in the concentration range 1 nM-10 microM. The 5-HT(3) receptor antagonist, ondansetron, increased the interval between CMMCs in the concentration range 100 nM-1 microM, whilst the 5-HT(1) receptor antagonist, methiothepin, the 5-HT(2) receptor antagonist, cyproheptadine and the 5-HT(4) receptor antagonist, SDZ 205 557, had no significant effects on the interval between CMMCs in the concentration range 1 nM-10 microM. The effects of 5-HT did not appear to be altered by the presence of ondansetron (1 microM) or cyproheptadine (1 microM). However, in the presence of ondansetron (1 microM), the further addition of cyproheptadine (1 microM) effectively abolished CMMCs. Furthermore, in the combined presence of these antagonists the effects of 5-HT were severely diminished. It is suggested that the frequency of CMMCs may be under the influence of endogenously released 5-HT in this preparation

4-Aminobenzoic Acid↗

PubMed1998

Evidence that myoelectric complexes in the isolated mouse colon may not be of myogenic origin.

The hypothesis that spontaneous depolarisations (myoelectric complexes, MCs) can occur in the absence of neuronal activity, depending on the level of the membrane potential, was systematically studied. In control Krebs' solution, MCs were recorded approximately every 5 min and were abolished by tetrodotoxin (TTX, 1.6 microM). However, TTX also induced sustained membrane depolarisation (19 mV) in the circular muscle. To test whether MCs were blocked by the depolarisation induced by TTX, graded membrane repolarisations were generated, in the continuing presence of TTX, using sodium nitroprusside (SNP, 10 nM-1 microM). Under these conditions, MC activity was not restored. The addition of SNP (1 microM) to control preparations, in normal Krebs' solution, hyperpolarised the membrane of the circular muscle cells, but did not inhibit ongoing MC activity. It is suggested that the underlying mechanisms involved in MC generation are unlikely to be dependent upon the level of membrane potential in circular smooth muscle.

Animals↗

PubMed1998

Disinhibition during myoelectric complexes in the mouse colon.

Intracellular microelectrodes were used to record electrically evoked inhibitory junction potentials (IJPs) and electrotonic potentials during spontaneous cyclical depolarisations (myoelectric complexes, MCs) in the circular muscle layer of mouse colon in vitro. In the presence of nifedipine (1-2 microM) and atropine (1 microM), MCs were recorded every 264 +/- 18 s. Between MCs, single electrical stimuli (15 V, 0.6 ms, every 8 s) elicited IJPs whose amplitudes remained constant. In comparison, during the depolarising phase of MCs, the mean IJP amplitude was reduced by 61 +/- 7%, while during the late plateau and early repolarising phase of MCs, IJP amplitude was increased (up to 20%). NG-nitro-L-arginine (NOLA, 100 microM) abolished the repolarisation phase between MCs, so that the circular muscle remained depolarised and the amplitude of MCs was reduced by 73 +/- 6%. However, the amplitude of evoked IJPs was unaffected, as was the decrease in their amplitude during the depolarising phase of the residual MCs. In the presence of NOLA (100 microM), the further addition of apamin (250 nM) reduced the amplitude of evoked IJPs by approximately half. However, the amplitudes of NOLA- and apamin-resistant IJPs were also attenuated by 82 +/- 5% during the depolarising phase of residual MCs (amplitude: 1.9 +/- 1 mV). However, during this phase, the amplitude of an electrotonic potential (evoked by extracellular current application) was not attenuated. Addition of hexamethonium (500 microM), or tetrodotoxin (TTX) (1.6 microM) to solutions containing NOLA and apamin were without effect on membrane potential, but the residual MCs and the cyclical attenuation in IJP amplitude were abolished. During the intervals between MCs, membrane potential is maintained under tonic inhibition, via spontaneous release of inhibitory neurotransmitter(s), predominantly through nitrergic mechanisms. The cyclical attenuation in the amplitude of the non-nitrergic IJP does not arise from cyclical postjunctional changes in membrane resistance or potential. Moreover, the generation of the depolarising phase of MCs involves the simultaneous suppression of both nitrergic and non-nitrergic inhibitory neurotransmission. It is suggested that MCs arise from presynaptic suppression of ongoing inhibitory neurotransmitter release.

Animals↗

PubMed1998

Inhibitory neurotransmission in the circular muscle layer of mouse colon.

Intracellular electrophysiological techniques were used to record the spontaneous myoelectric activity in the circular muscle layer of an in vitro preparation of whole mouse colon. In 34 out of 58 preparations, spontaneous depolarisations (myoelectric complexes, MCs) were recorded cyclically, about every 4 min. In these preparations, apamin (250 nM) and NG-nitro-L-arginine (NOLA, 100 microM) depolarised the membrane potential between MCs by 8 mV or 13 mV, respectively. Tetrodotoxin (1.6 microM) abolished MCs and also induced depolarisation (17 mV). In the remaining 24 preparations, MCs were not recorded and the membrane potential was significantly depolarised compared to the membrane potential between MCs. NOLA (100 microM), apamin (250 nM) and tetrodotoxin (1.6 microM) were without significant effect on membrane potential. It is suggested that in preparations that exhibit MC cycling, membrane potential between MCs is maintained in a state of tonic inhibition, predominantly mediated by nitrergic mechanisms generated via spontaneously active inhibitory neurons. Apamin-sensitive channels may also be involved in the inhibition.

Animals↗

PubMed1998

Spontaneous and evoked inhibitory junction potentials in the circular muscle layer of mouse colon.

Intracellular microelectrodes were used to record spontaneous and evoked inhibitory junction potentials (IJPs) from the circular muscle layer of an in vitro preparation of whole mouse colon. Membrane potential recordings were made from cells of the mid to distal region of colon at 36 +/- 1 degrees C in a modified Krebs' solution that contained atropine (1 microM) and nifedipine (1-2 microM). Spontaneously occurring hyperpolarisations of irregular amplitude and frequency (range: up to 20 mV and 2 Hz) were recorded that were resistant to N(G)-nitro-L-arginine (NOLA, 100 microM), but were abolished by tetrodotoxin (TTX, 1.6 microM) or apamin (250 nM). These were considered to be spontaneous IJPs as a consequence of activity in inhibitory motor neurons. Single electrical stimuli (0.6 ms, 15 V), elicited a fast IJP, whose time course could be superimposed on spontaneous IJPs of similar amplitude. The amplitude of evoked IJPs was not depressed by NOLA (100 microM). However, in NOLA (100 microM), further addition of apamin (250 nM) significantly depressed the amplitudes of the evoked IJPs by 44%. NOLA- and apamin-resistant evoked IJPs were abolished by TTX (1.6 microM). It is suggested, that in the circular muscle layer of mouse colon, NO does not mediate the fast hyperpolarisations associated with spontaneous or evoked IJPs. Apamin abolished spontaneous IJPs, but electrical stimuli evoked an IJP with apamin-sensitive and resistant components both of which were non-nitrergic in origin.

Animals↗

PubMed1998

Inhibition of voltage-activated K+ currents in smooth muscle cells of the guinea pig proximal colon by noradrenergic agonists.

1. The effects of noradrenaline and isoprenaline on the Ca2+i-insensitive, voltage-activated K+ current in smooth muscle cells from the circular muscle layer of the guinea pig proximal colon were investigated by using standard whole-cell patch-clamp techniques at room temperature (22-24 degrees C). 2. The Ca2+-activated K+ current was eliminated by bathing cells in tetraethylammonium (TEA;2-5 mM) and a Ca2+-entry blocker (Cd2+, 0.1 mM) or nifedipine, 2-10 microM) and by internally perfusing cells with 3 mM EGTA. 3. Two Ca2+i-insensitive, voltage-activated K+ currents were recorded at potentials positive to -50 mV: (a) a transient K+ current (IKto) that was blocked by 4-aminopyridine (5 mM) and (b) a delayed rectifier-type K+ current (IKdel) that was blocked by TEA (>10 mM). 4. Both noradrenaline (10-50 microM) and isoprenaline (5-50 microM) reduced the amplitudes of IKto and IKdel irreversibly after a slow onset (2-5 min). This reduction was mimicked by forskolin (50-100 microM) and by 8 bromo-c-AMP (500 microM). 5. The voltage of half-maximal availability (V0.5) of IKto (-74.6+/-2.3 mV) was unaffected by isoprenaline (10 microM) (-76.7+/-3.6 mV, n=4), but the background "leak" current (Ileak) was increased from -48+/-9 to -70+/-20 pA. 6. Our data suggest that stimulation of beta-adrenoceptors in the circular muscle layer of the guinea pig proximal colon inhibits voltage-activated Ca2+i-insensitive K+ currents.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1998

Effects of sympathetic nerve stimulation on membrane potential in the circular muscle layer of mouse distal colon.

Little is known about the effects of sympathetic nerve stimulation on the membrane potential of colonic smooth muscle. In the distal colon of the mouse, intracellular microelectrodes were used to record the effects of lumbar colonic (LCN) and intermesenteric nerve (IMN) stimulation on circular muscle membrane potential in vitro. A two-compartment organ bath was used to selectively perfuse the colon and inferior mesenteric ganglion (IMG). In the presence of nifedipine (1-2 microM) (to the colonic compartment only), spontaneous depolarizations (myoelectric complexes, MCs) were recorded about every 4 min. MCs consisted of a prolonged burst of rapid oscillations in membrane potential (approximately 2 Hz) that were superimposed on a slow depolarization (mean amplitude 12 mV). Single electrical stimuli (600 microseconds duration) delivered to the LCN or IMN did not elicit a detectable change in the membrane potential. However, trains of stimuli (e.g., 60 pulses at 10-20 Hz) to the LCN or IMN during the intervals between MCs evoked a depolarization (with superimposed action potentials in the absence of nifedipine). Trains of stimuli delivered during the plateau phase of the MC reduced or abolished the rapid oscillations, without a further membrane depolarization. The MC period was unaffected by stimulation of the IMN or LCN. Responses were abolished by the selective perfusion of guanethidine (1 microM) to the colon, or by severing the LCN. Hexamethonium (500 microM) (to the colon) abolished MCs, induced sustained depolarization and attenuated the amplitude of the sympathetic depolarizations by 74%. In hexamethonium, sympathetic responses remained attenuated when the membrane of the circular muscle was repolarised by sodium nitroprusside (1 microM). Immunohistochemical studies of the colon revealed intense immunoreactivity for tyrosine hydroxylase in the myenteric plexus but not in the circular muscle layer. It is suggested that responses to sympathetic nerve stimulation in the circular muscle layer of the mouse colon are secondary to actions on the enteric nervous system.

Animals↗

PubMed1998

Second-, minute- and hour-metronomes of intestinal pacemakers.

1. Movements of the gastrointestinal tract are required for the digestion of food and the expulsion of waste products. 2. The present paper will discuss the nature of electrical rhythms underlying some intestinal motility patterns. 3. The rhythms are generated by pacemakers with cycle rates appropriate to controlling individual contractions, motor patternings or switching between different motor programmes. 4. Electrical rhythms are discussed with periods of the order of seconds, minutes and hours. 5. Particular discussion is centred on rhythms recorded from the small and large intestine of the mouse.

Animals↗

PubMed1997

Computer simulation of the enteric neural circuits mediating an ascending reflex: roles of fast and slow excitatory outputs of sensory neurons.

Recent electrophysiological studies of the properties of intestinal reflexes and the neurons that mediate them indicate that the intrinsic sensory neurons may transmit to second order neurons via either fast (30-50 ms duration) or slow (10-60 s duration) excitatory synaptic potentials or both. Which of these possible modes of transmission is involved in the initiation of motility reflexes has not been determined and it is not clear and what the consequences of the different forms of synaptic transmission would be for the properties of the reflex pathways. In the present study, this question has been addressed by the use off a suite of computer programs, Plexus, which was written to simulate the activity of the neurons of the enteric nervous system during intestinal reflexes. The programs construct a simulated enteric nerve circuit based on anatomical and physiological data about the number, functions and interconnections of neurons involved in the control of motility. The membrane potentials of neurons are calculated individually from physiological data about the reversal potentials and membrane conductances for Na+, K+ and Cl-. Synaptic potentials are simulated by changes in specific conductances based on physiological data. The results of each simulation are monitored by recording the membrane potentials of up to 16 separate defined neurons and by recording the summed activity of whole classes of neurons as a function of time and location in the stimulated network. The present series of experiments simulated the behaviour of a network consisting of 18,898 sensory neurons and 3708 ascending interneurons after 75% of the sensory neurons lying in the anal 10 mm of a 30 mm long segment of small intestine were stimulated once. The results were compared with electrophysiological data recorded from myenteric neurons during ascending reflexes evoked either by distension or mechanical stimulation of the mucosa. When transmission from sensory neurons to ascending interneurons was via fast excitatory synaptic potentials, the latencies and durations of the simulated responses were too brief to match the electrophysiologically recorded responses. When transmission from sensory neurons was via slow excitatory synaptic potentials, the latencies were very similar to those recorded physiologically, but the durations of the stimulated responses were much longer than seen in physiological experiments. The latencies and durations of simulated and physiologically recorded responses matched only when the firing of ascending interneurons was limited to the beginning of a slow excitatory synaptic (in this study by limiting the duration of the decrease in K+ conductance). The simulation provided several physiologically testable predictions, indicating that Plexus is an important tool for the investigation of the properties and behaviour of the enteric nervous system.

Computer Simulation↗

PubMed1997

Colonic migrating motor complexes (CMMCs) in the isolated mouse colon.

Spontaneous contractions were recorded from the circular muscle layer at three sites along the isolated mouse colon. The interval between contractions was approximately 4.5 min. The mean duration of the contractions ranged from 26 sec in the distal colon to 45 sec in the proximal colon. Contractions migrating more than half the length of the colon were termed colonic migrating motor complexes (CMMCs). Over 90% of tissues demonstrated migration predominantly in an aboral direction. Hyoscine (10(-6) M) decreased the amplitude of the CMMCs by at least 40% but had no significant effect on the interval or duration of the CMMCs. Nifedipine (10(-6) M) significantly decreased the amplitude of the CMMCs by 95% but did not alter the duration or the interval between the CMMCs. Hexamethonium (5 x 10(-4) M) and tetrodotoxin (TTX; 2 x 10(-6) M) abolished all CMMC activity. TTX increased the resting tone of the preparations. Nitro-L-arginine (10(-4) M) increased the resting tone of the preparations and significantly decreased the interval between the CMMCs by approximately 80% but had no significant effect on the duration of the CMMCs. The results suggest CMMCs migrate predominantly in an aboral direction and are neurogenic in origin. Nitric oxide may be involved in maintaining inhibition of the muscle between CMMCs.

Animals↗

PubMed1996

Characterization of the membrane conductance changes underlying the apamin-resistant NANC inhibitory junction potential in the guinea-pig proximal and distal colon.

The nature of the electrically- or stretch-evoked nonadrenergic, noncholinergic (NANC) inhibitory junction potentials (IJPs) in circular smooth muscle cells of the guinea-pig proximal and distal colon were investigated using standard intracellular microelectrode recording techniques. We have confirmed that the NANC IJP, recorded in the presence of hyoscine (1 microM) and nifedipine (1 microM), can be divided into two components with apamin (250 nM), a blocker of the small conductance Ca2(+)-activated K+ channels. Both the apamin-sensitive and the apamin-resistant components of the IJP were blocked by tetrodotoxin (1.6 microM) or by lowering the external Ca2+ concentration (to 0.25 mM). The apamin-sensitive IJP was also blocked by omega-conotoxin GVIA (100 nM), a blocker of 'N-type' Ca2+ channels. The apamin-resistant IJP and rebound post-stimulus depolarization (PSD) were reduced upon exposure to either NG-L-arginine (NOLA), an inhibitor of nitric oxide synthase (NOS), or the nitric oxide (NO) scavenger, haemoglobin. The effects of NOLA were partially reversed in the presence of excess L-arginine, a substrate for NOS, suggesting that NO, or a related NO-donor compound, is likely to be the apamin-resistant inhibitory transmitter. Blockade of either the apamin-sensitive or apamin-resistant IJP was associated with membrane depolarization and a decrease in the membrane conductance in the absence of nerve stimulation. In the proximal colon, the apamin-resistant IJP and PSD could both be demonstrated to arise from an increase in the membrane conductance after subtraction of a non-linear background conductance. The hyperpolarization upon repetitive NANC nerve stimulation was mimicked by the NO donor, S-nitroso-L-cysteine (2.5-25 microM), which evoked a transient apamin-sensitive, but omega-conotoxin GVIA resistant, component followed by a slower apamin-resistant component. These results suggest that neurally-released NO has a number of actions in the guinea-pig colon, causing apamin-resistant hyperpolarization and depolarization, as well as directly opening apamin-sensitive K+ channels.

Animals↗

PubMed1996

Effects of nitric oxide (NO) and NO donors on the membrane conductance of circular smooth muscle cells of the guinea-pig proximal colon.

1. The membrane conductance changes underlying the membrane hyperpolarizations induced by nitric oxide (NO), S-nitroso-L-cysteine (NC) and sodium nitroprusside (SNP) were investigated in the circular smooth muscle cells of the guinea-pig proximal colon, by use of standard intracellular microelectrode recording techniques. 2. NO (1%), NC (2.5-25 microM) and SNP (1-1000 microM) induced membrane hyperpolarization in a concentration-dependent manner, the hyperpolarizations to NO and NC developing more rapidly than those to SNP. The slower-developing responses to SNP were mimicked by the membrane permeable analogue of guanosine 3':5' cyclic-monophosphate (cyclic GMP), 8-bromo-cyclic GMP (500 microM), and by isoprenaline (10 microM). 3. The hyperpolarizations to NC and SNP were reduced in a low Ca2+ (0.25 mM) saline and upon the addition of haemoglobin (20 microM), but were not effected by NG-nitro-L-arginine (L-NOARG) (100 microM) or omega-conotoxin GVIA (100 nM). the hyperpolarizations to SNP were also significantly reduced by methylene blue (50 microM). 4. Apamin (250 nM) depolarized the membrane potential approximately 10 mV and reduced the initial transient component of the hyperpolarization to NO (1%) and NC (25 microM), but had no effects on the hyperpolarizations to SNP and cyclic GMP. Tetraethylammonium (TEA) (5-15 mM), had little effect on the membrane responses to NO(1%), NC(2.5-25 microM), SNP(100(-1000) microM) or cyclic GMP(500 microM). However, TEA (5-15 mM) reduced the membrane hyperpolarizations to SNP (10 microM) and isoprenaline (10 microM) in a concentration-dependent manner. The hyperpolarization to isoprenaline (10 microM) remaining in the presence of 15 mM TEA was blocked by ouabain (10 microM). 5. The amplitude of electronic potentials (1 s duration) elicited during NO donor hyperpolarizations were little changed or only slightly reduced (5-25%). However, the amplitude of the electrotonic potentials elicited during maintained electrically-induced hyperpolarizations of similar amplitude were significantly increased (30-150%), suggesting that the non-linear membrane properties of the proximal colon partially mask an increase in membrane conductance elicited during the NO donor hyperpolarizations. 6. Membrane hyperpolarization in the presence of an NO donor, 8-bromo-cyclic GMP, isoprenaline, or upon application of a maintained hyperpolarizing electrical current, often evoked oscillations of the membrane potential. These oscillations were prevented by Cs+ (1 mM). 7. These results indicate that NO and NC hyperpolarize the circular muscle of the proximal colon by activating at least two TEA-resistant membrane K+ conductances, one of which is sensitive to apamin blockade. The K+ conductance increases activated by SNP or 8-bromo-cyclic GMP were little effected by apamin, perhaps suggesting a common mechanism. In contrast, the hyperpolarization to isoprenaline appears to involve the activation of TEA-sensitive Ca2(+)-activated K+ ('BK') channels, as well as a Na:K ATPase. Finally, the 'background' membrane conductance of the circular muscle cells of the proximal colon decreased upon membrane hyperpolarization to reveal oscillations of the membrane potential which may well represent 'pacemaker' or 'slow wave' activity.

Animals↗

PubMed1995

Neurogenic control of myoelectric complexes in the mouse isolated colon.

BACKGROUND/AIMS: Little is known about the mechanisms controlling colonic migrating electrical activity. This study investigates the neural processes involved in the generation of migrating myoelectric complexes in the isolated mouse colon. METHODS: Intracellular electrophysiological recordings were obtained from the circular muscle layer of the mouse colon in vitro in the presence of 2 mumol/L nifedipine. RESULTS: Complexes occurred approximately every 3 minutes and consisted of 1 mumol/L hyoscine-sensitive rapid oscillations (approximately 2 Hz) superimposed on a slow depolarization (approximately 17 mV); the latter was often preceded by a precomplex hyperpolarization (approximately 7 mV) that was reduced by 250 nmol/L apamin. Five hundred micromolars of hexamethonium or 2 mumol/L of tetrodotoxin abolished the complexes and depolarized the muscle by 8.7 +/- 1.3 mV (n = 9) or 12.1 +/- 1.4 mV (n = 5), respectively. Carbachol (50 nmol/L to 5 mumol/L) produced dose-dependent depolarizations but without rapid oscillations. The nitric oxide synthase inhibitor NG-nitro-L-arginine (100 mumol/L) depolarized the tissue by 17.2 +/- 1.6 mV (n = 8) but had no effect on the rapid oscillations. In the presence of 2 mumol/L tetrodotoxin, 5 mumol/L sodium nitroprusside produced a sustained hyperpolarization (15.5 +/- 2.0 mV; n = 5) but did not restore complexes. CONCLUSIONS: In the isolated mouse colon, the membrane potential between complexes is maintained by the release of inhibitory neurotransmitters (including nitric oxide), and the formation of complexes involves disinhibition and the simultaneous activation of cholinergic motor nerves.

Animals↗

PubMed1993

Voltage-gated ionic currents in smooth muscle cells of guinea pig proximal colon.

Smooth muscle cells enzymatically dispersed from the circular muscle layer of the guinea pig colon were examined for the expression of voltage-gated ionic currents using the whole cell patch-clamp technique. Three outward currents and one inward current were identified and characterized. The inward current, at physiological potentials, was generated by a voltage-gated Ca2+ conductance that was permeable to Ba2+ but blocked by Cd2+ (0.1 mM) or nifedipine (10 microM). The three outward currents were carried by K+, abolished when cells were internally perfused with Cs+, and distinguished by their sensitivity to known K(+)-channel blockers. A K+ current dependent on Ca2+ entry was activated at potentials positive to -40 mV and abolished by low concentrations (2-5 mM) of tetraethylammonium (TEA). A delayed rectifier-type K+ current activated slowly at potentials positive to -30 mV, showed little inactivation, and was abolished by higher concentrations (> 5 mM) of TEA. A rapidly activating and inactivating transient outward K+ current (IKto), activated at potentials positive to -60 mV, was abolished by low concentrations (3-5 mM) of 4-aminopyridine and was relatively insensitive to TEA (10-126 mM) blockade. The overlapping steady-state activation and inactivation curves of IKto revealed a "window current" phenomenon between -60 and -40 mV. The rapid activation of IKto at membrane potentials more negative than those for Ca2+ current suggests that IKto will influence cell excitability by delaying the upstroke of the action potential. The present data provide an ionic basis for the regenerative excitatory activity in the circular muscle layer of the guinea pig proximal colon.

4-Aminopyridine↗

PubMed1992

Effects of a nitric oxide synthase inhibitor on non-cholinergic junction potentials in the circular muscle of the guinea pig ileum.

Intracellular microelectrodes were used to record junction potentials from the circular muscle cells of the guinea pig ileum in vitro at 37 degrees C in a modified Krebs solution containing nifedipine (1-2 microM) and hyoscine (1 microM). Transmural nerve stimulation, using volleys of three pulses at 50 Hz, produced a complex response consisting of an inhibitory junction potential (IJP) followed by a prolonged depolarization. Following the addition of the nitric oxide synthase inhibitor NG-nitro-L-arginine (NOLA, 100 microM) the amplitude of the IJP (recorded 10 mm aboral to the stimulating electrodes) was increased by approx. 10% (n = 4). The further addition of apamin (250 nM) abolished the IJP revealing a non-cholinergic excitatory junction potential (EJP). In other experiments (n = 8), preparations were treated with apamin then subjected to substance P desensitization (500 nM, > 20 min). Transmural nerve stimulation now produced a triphasic response (recorded 1 mm aboral to the stimulating electrodes) consisting of: (a) an initial hyperpolarization (approx. 5 mV) lasting about 1 s; followed by (b) a depolarization reaching a peak (approx. 7 mV less negative than the RMP) approx. 2 s after nerve stimulation; and finally (c) a small (approx. 3 mV) hyperpolarization. The addition of NOLA reduced all three phases by 80-90% (n = 8). The subsequent addition of L-arginine (5 mM) partially reversed these effects (n = 3). Conditioning hyperpolarization up to 20 mV increased the amplitude of the NOLA-sensitive IJP and EJP. Further conditioning hyperpolarization reduced the amplitude of the IJP and enhanced the amplitude of the EJP. Large conditioning hyperpolarizations (> 60 mV) reduced the amplitude of both the IJP and EJP. An estimation of the membrane conductance changes occurring during the initial hyperpolarization and depolarization suggest that it was either unchanged or increased. During large conditioning hyperpolarizations in the absence of nerve stimulation, the membrane potential was unstable and began to show spontaneous oscillations (up to 30 mV, every 4-5 s) resembling slow waves. These experiments indicate that NO, or a related compound, appears to mediate the nerve induced apamin-resistant IJP and substance P- and hyoscine-resistant EJP in the circular muscle of the guinea pig ileum.

Amino Acid Oxidoreductases↗