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M Neunlist

Publications and source records attributed to M Neunlist.

26 records · Page 2Linked to original sources
PubMed1998

Polarized enteric submucosal circuits involved in secretory responses of the guinea-pig proximal colon.

1. Neuronal retrograde tracing with the dye DiI (1,1'-didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), in combination with immunohistochemical detection of choline acetyltransferase (ChAT) and vasoactive intestinal peptide (VIP), were used to identify the innervation of the mucosa of the guinea-pig proximal colon by submucosal neurones. Ussing chamber experiments were performed to measure changes in short circuit current (delta Isc) evoked by electrical stimulation of the oral or anal end of the preparation. 2. The tracing studies revealed that the mucosa was primarily innervated by descending neurones (78%); the vast majority of these were VIP positive (85%). The numerically smaller ascending pathway (13%) was predominantly ChAT positive (69%). A small population (9%) of DiI-labelled neurones projected circumferentially. 3. Ussing chamber experiments revealed that oral electrical stimulation induced a significantly larger delta Isc than anal stimulation. The VIP antagonist VIP(6-28) significantly reduced only orally induced delta Isc. Anally induced delta Isc were significantly more atropine sensitive that orally induced delta Isc. Tissue incubation with carbachol or VIP significantly potentiated delta Isc induced by VIP and carbachol, respectively, indicating cross-potentiation. 4. This study provides the first functional demonstration of polarized innervation patterns from submucosal neurones to enteric mucosa. The ascending ChAT and descending VIP pathways suggest the existence of reflexes resulting in preferential release of VIP or acetylcholine. The distinct pathways might favour the observed cross-potentiation of cholinergic and VIPergic mediated secretion.

Action Potentials↗
PubMed1998

Post- and presynaptic effects of norepinephrine in guinea-pig colonic submucous plexus.

Intracellular recording techniques were used to investigate the effects of norepinephrine on submucous neurones in the guinea-pig distal colon. In 81% of the neurones, pressure microejection of norepinephrine produced a membrane hyperpolarization associated with a decrease in excitability and input resistance. Microejection of clonidine (1 microM) mimicked the norepinephrine-induced hyperpolarization, whereas both phentolamine (1 microM) and yohimbine (1 microM) reversibly suppressed it. Superfusion of norepinephrine (1 nM - 10 microM) hyperpolarized the cells in a concentration-dependent manner. Norepinephrine and clonidine (1 nM - 10 microM) caused a concentration-dependent presynaptic inhibition of stimulus-evoked cholinergic fast excitatory postsynaptic potential. Slow inhibitory post-synaptic potentials (sISPSs) were induced by focal electrical stimulation of the interganglionic fibre tracts in 43% of the neurones tested. Superfusion of both phentolamine (1 microM) and yohimbine (1 microM) reduced the sIPSPs while prazosin (1 microM) had no significant effect. We concluded that norepinephrine acted post- and presynaptically via alpha 2-adrenoreceptors to have an inhibitory effect on the guinea-pig colonic submucous. In addition, our study strongly supported the role of norepinephrine as a mediator of the sIPSPs. As a result, norepinephrine would primarily suppress information transfer within the neuronal circuits in guinea-pig colonic submucosal plexus.

Acetylcholine↗
PubMed1997

Projections and neurochemical coding of myenteric neurons innervating the mucosa of the guinea pig proximal colon.

Myenteric neurons projecting to the mucosa of the guinea pig proximal colon were identified using the combination of a neuronal tracing method and immunohistochemical techniques. The tracer DiI (1, 1'didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate) was applied onto the mucosa of a specimen of proximal colon which was then placed in organotypic culture to allow retrograde transport of the dye. After culture, the myenteric plexus was stained with antisera raised against choline acetyltransferase (ChAT) and calbindin (Calb). Of the myenteric neurons labeled with DiI, 99% had smooth cell bodies with Dogiel Type II morphology. Of these neurons, 70% projected in the longitudinal direction and the majority of them (65%) were located anally from the DiI application site, i.e., had ascending projections. Ascending neurons projected over significantly shorter distances than descending ones (3.1+/-0.5 mm vs. 4.6+/-1.2 mm, respectively; P<0.01). Of the labeled myenteric neurons, 98% were ChAT immunoreactive. Of these neurons, 78% were also immunoreactive for Calb and were preferentially ascending neurons. ChAT-immunoreactive but Calb-negative neurons did not have preferential projection. This study revealed the presence of two populations of myenteric neurons projecting to the mucosa of the guinea pig proximal colon. Morphological characteristics and neurochemical coding were suggestive for a putative sensory function for these neurons.

Animals↗
PubMed1997

Dose-dependent reduction of cardiac transmembrane potential by high-intensity electrical shocks.

Cardiac tissue dysfunction can result from high-intensity electrical shocks and is manifested as changes in transmembrane potential (Vm). Ten-millisecond shock pulses (SPs) of varying intensity and polarity were applied to frog ventricle in diastole, and Vm was quantified directly under the stimulating electrode by an optical method using voltage-sensitive dye. As SP intensities were increased, the shock-induced action potential (AP) plateau and AP amplitude (APAs) decreased sigmoidally toward 75-85% of the control AP amplitude (APAc) and zero, respectively. APAs was shifted toward lower current densities for anodal compared with cathodal SPs (half-maximal values 185 and 238 mA/cm2, respectively; P = 0.02). Recovery of APAs was marginally significant 1 s after SP delivery (P = 0.063). The peak change in Vm during SP (across all intensity levels) was -200% APAc for anodal and +125% APAc for cathodal pulses. In conclusion, we show that SP reduces APA in a sigmoidal fashion at strengths > 10-20 x diastolic threshold and is more deleterious for anodal polarities.

Action Potentials↗
PubMed1995

Spatial distribution of cardiac transmembrane potentials around an extracellular electrode: dependence on fiber orientation.

Recent theoretical models of cardiac electrical stimulation or defibrillation predict a complex spatial pattern of transmembrane potential (Vm) around a stimulating electrode, resulting from the formation of virtual electrodes of reversed polarity. The pattern of membrane polarization has been attributed to the anisotropic structure of the tissue. To verify such model predictions experimentally, an optical technique using a fluorescent voltage-sensitive dye was used to map the spatial distribution of Vm around a 150-microns-radius extracellular unipolar electrode. An S1-S2 stimulation protocol was used, and vm was measured during an S2 pulse having an intensity equal to 10x the cathodal diastolic threshold of excitation. The recordings were obtained on the endocardial surface of bullfrog atrium in directions parallel and perpendicular to the cardiac fibers. In the longitudinal fiber direction, the membrane depolarized for cathodal pulses (and hyperpolarized for anodal pulses) but only in a region within 445 +/- 112 microns (and 616 +/- 78 microns for anodal pulses) from the center of the electrode (n = 9). Outside this region, vm reversed polarity and reached a local maximum at 922 +/- 136 microns (and 988 +/- 117 microns for anodal pulses) (n = 9). Beyond this point vm decayed to zero over a distance of 1.5-2 mm. In the transverse fiber direction, the membrane depolarized for cathodal pulses (and hyperpolarized for anodal pulses) at all distances from the electrode. The amplitude of the response decreased with distance from the electrode with an exponential decay constant of 343 +/- 110 microns for cathodal pulses and 253 +/- 91 microns for anodal pulses (n = 7). The results were qualitatively similar in both fiber directions when the atrium was bathed in a solution containing ionic channel blockers. A two-dimensional computer model was formulated for the case of highly anisotropic cardiac tissue and qualitatively accounts for nearly all the observed spatial and temporal behavior of vm in the two fiber directions. The relationships between vm and both the "activating function" and extracellular potential gradient are discussed.

Animals↗
PubMed1994

Optical recordings of ventricular excitability of frog heart by an extracellular stimulating point electrode.

To enhance understanding of the excitability of cardiac muscle during rest, an optical technique using the fluorescent voltage sensitive dye di-4-ANEPPS was used. Unlike conventional electrical recordings, optical recordings are free from electrical artifacts and, therefore, allow the observation of the transmembrane potential not only following the stimulation pulse, but also during the pulse itself. Transmembrane potentials (Vm) were recorded optically from frog ventricular epicardium in calcium containing Ringer's solution directly under an extracellular stimulating point electrode. Anodal and cathodal S1 stimuli were applied at rest. As observed by previous investigators, the post-pulse excitatory responses for cathodal pulses, compared with anodal pulses were greater. Changes in transmembrane potential (delta Vm) during the pulse were as expected for a passive cable only for low intensity pulses (< 4 x the cathodal threshold of excitation in diastole, CTE). However, at the higher intensities necessary to produce an excitatory response (> 6-8 x CTE), an "irregular" response in Vm was observed--a reversal of the hyperpolarization during an anodal stimulus pulse and a reversal of the depolarization during a cathodal stimulus pulse. To elucidate further the biophysical basis for this behavior, delta Vm was mapped around the stimulating electrode. During stimulation, regions could be observed having a response with opposite polarity to that under the electrode (i.e., depolarization for an anodal pulse and hyperpolarization for a cathodal pulse). Removal of the bath solution or the addition of channel blockers did not eliminate the occurrence of these regions. These regions appear to be the basis for the irregular behavior of delta Vm directly under the electrode as well as for anodal excitation.

Animals↗
PubMed1992

Design and use of an "optrode" for optical recordings of cardiac action potentials.

An optical method was used to measure action potentials from frog ventricle, in vitro, under normal physiological conditions with 0.5-1 mM Ca2+ Ringer's solution. The approach presented in this paper involves a portable fluorimeter coupled to a multimode optical fiber running into a glass pipette ("optrode") to carry both excitation light to and fluorescence from the ventricle stained with the voltage sensitive dye di-4-ANEPPS. A suction technique was used to stabilize the optrode-tissue interface, significantly reducing motion artifacts from the beating ventricle. The typical fractional change in fluorescence intensity for an action potential was -9%. The optical recordings faithfully reproduced membrane action potentials as measured with microelectrode recordings. To confirm further the validity of our method we studied the effect of an increasing stimulation rate on the optical action potential. The amplitude of the action potential did not increase, and the change in action potential duration was similar to published results obtained with microelectrode recordings, suggesting that our optical action potentials are relatively free of motion artifacts. Finally, our optical recordings suggest that during anodal and cathodal point stimulation, the time course of membrane potential differs from that predicted simply by a passive cable model.

Action Potentials↗