Biomedical subjects

T M Dwyer

Publications and source records attributed to T M Dwyer.

At least 37 records · Page 2Linked to original sources

PubMed1994

Faecal dry weight and potassium are related to faecal sodium and plasma aldosterone in rats chronically fed on varying amounts of sodium or potassium chlorides.

Recent studies have shown that faecal residue (dry weight) and Na and K increase with increasing levels of dietary fibre, an effect which may be related to unstirred layers that slow absorption and the flow rate of chyme through the gastrointestinal tract. Salts of Na are the primary osmotic components of chyme and influence both retention of fluid in the bowel and transit of fluid from the small to the large intestine. The present study examines the chronic effects of dietary Na and K intake on faecal Na, K and residue excretion. Male Sprague-Dawley rats were given 12-13 g feed/d (control (g/kg): Na 4, K 8.5) for 1 week, followed by a 4-week period where Na or K intake was altered (0.01-3 times control levels). These diets altered chronic (> 1 week) faecal residue excretion and affected Na and K excretion by 8-, 310- and 2100-fold respectively. Low dietary Na reduced faecal Na and residue; K excretion was doubled during week 1, but fell over weeks 2-4 despite a 4-5-fold increase in plasma aldosterone. Chronic high dietary Na increased faecal Na, residue and K despite a 60% decrease in plasma aldosterone. Chronic low dietary K decreased faecal Na, K and residue and plasma aldosterone. Chronic high dietary K did not alter faecal Na and K despite increased faecal residue and a 4-5-fold increase in plasma aldosterone. Faecal water was unchanged by diet, paralleling changes in faecal residue. Analyses of the results provide systematic models of chronic regulation of faecal Na, K and residue excretions. When plasma aldosterone is low (< 160 ng/l), weekly faecal residue excretion is equal to 3.6 g (fibre intake was 2.6-2.7 g/week)+2.6 g/mmol Na, and K excretion is equal to 0.55 mmol/mmol Na. When plasma aldosterone is high (> 1500 ng/l) weekly faecal residue excretion is equal to 2.3 g + 2.6 g/mmol Na, and K excretion is equal to 0.7 mmol/week + 0.55 mmol/mmol Na.

Aldosterone↗

PubMed1994

Basal and thrombin-stimulated platelet cell calcium.

To study platelet cell calcium in dogs, we measured the fura-2-determined calcium concentration (Cai) in platelets derived from blood drawn through indwelling arterial catheters in four dogs. Platelet samples were obtained 3 days a week for 39 days. Canine platelet Cai averages 40.5 +/- 1.7 nmol/L when corrected for extracellular fura-2 dye (n = 46); this mean is significantly smaller than the value obtained without such a correction (50.2 +/- 2.0 nmol/L, P < .05). The amplitude of thrombin-induced Ca2+ transients is proportional to basal Cai, increasing 4.9 +/- 0.9 nmol/L peak Ca2+ per nmol/L increase in baseline Ca2+.

Animals↗

PubMed1993

Aldosterone secretion and the mechanism of potassium adaptation in rats.

In order to better understand the cellular mechanism of potassium (K+) adaptation, the sensitivity of aldosterone secretion to acute changes in extracellular K+ concentration was studied in freshly dissected adrenal capsules of rats adapted to diets of high or low K+ content, of rats adapted to low or high sodium (Na+) diets, and also of control rats. In control tissues, the aldosterone secretion from the capsules of an individual animal averages 0.44 +/- 0.05 nmol/h, increasing 4.4-fold between 2 and 8 mM K+ but decreasing between 8 and 10 mM K+. Although a high K+ diet increases aldosterone secretion by only 34% at 4 mM K+, the rate of secretion increases 3.7-fold more steeply than control as the K+ concentration increases. This change is equivalent to a parallel 3.1-fold increase in the effective number of T- and L-type calcium (Ca2+) channels, accompanied by a 1.3-fold increase in the K(+)-insensitive rate of aldosterone secretion. In contrast, after Na+ restriction, aldosterone secretion is about 3 times the control rate for all K+ concentrations tested, equivalent to an increase in the basal rate and the effective number of L-channels. Thus, the alteration in the number of effective T-channels is specific to diets of increased K+ content, not simply an effect of increased secretory capacity. After a low K+ diet, aldosterone secretion is 18% of control at 4 mM K+ and changes little with the K+ concentration, consistent with a 94% to 96% decrease in the effective number of T- and L-channels plus a 77% decrease in the K(+)-insensitive rate of aldosterone secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

PubMed1993

Two forms of mouse syntrophin, a 58 kd dystrophin-associated protein, differ in primary structure and tissue distribution.

Syntrophin, a 58 kd extrinsic membrane protein, is concentrated at postsynaptic sites at the neuromuscular junction and may be involved in clustering acetylcholine receptors. In muscle and nonmuscle tissues, syntrophin is associated with dystrophin, utrophin, and two homologs of the dystrophin carboxy-terminal region. We have isolated three cDNAs encoding Torpedo and mouse syntrophins. The Torpedo cDNA encodes a full-length protein, and on Northern blots recognizes a 3.5 kb mRNA. The two mouse syntrophin cDNAs are products of separate genes but encode proteins that share 50% identity. Syntrophin-1 mRNA (2.2 kb) is expressed at highest levels in skeletal muscle. Syntrophin-2 mRNAs (2.2, 5.0, and 10 kb) are expressed in all mouse tissues examined. These patterns of expression suggest that syntrophin-1 and syntrophin-2 may associate with different members of the dystrophin family.

Amino Acid Sequence↗

PubMed1992

Transient cholinergic glycoconjugate secretion from swine tracheal submucosal gland cells.

By inference, muscarinic stimulation of glycoconjugate release from tracheal submucosal gland cells appears to be a transient, nonequilibrium process (J. M. Farley and T. M. Dwyer, Life Sci. 48: 59-67, 1991). To directly characterize the release kinetics of glycoconjugate, we developed an enzyme-linked lectin assay (ELLA) of much improved precision and resolution. To collect secreted products with an improved time resolution, freshly isolated swine tracheal submucosal gland cells were continuously superfused with medium 199 at 37 degrees, buffered with N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) and CO2 or bicarbonate; fractions were collected every 15 s to 2 min. A 30-s pulse of 30 nM acetylcholine (ACh) increased the rate of glycoconjugate release by 10- to 25-fold for 2-3 min. The peak response averaged 14.2 +/- 9.0 ng protein.ml-1.min-1. 4 million cells-1 or 3.6 +/- 2.3 fg.cell-1.min-1 for 30 nM ACh and 16.2 +/- 3.0 ng protein.ml-1.min-1 for 100 nM ACh. There was no significant glycoconjugate release following a 30-s pulse of either 10 nM or 1 microM ACh. A second pulse after 7 min had no measurable effect on glycoconjugate release but a full response was obtained after 30 min. A continuous superfusion begun 1 min following the 30-s pulse resulted in a greater release of glycoconjugate than the pulse alone, but the response was not sustained, falling to twice basal levels within 5 min. We conclude that a brief muscarinic stimulation causes a triggered release of mucus glycoprotein followed by a relative refractory period.

Acetylcholine↗

PubMed1992

Cromakalim effects of acetylcholine-induced changes in cytosolic calcium and tension in swine trachealis.

The effects of cromakalim, an ATP-sensitive K+ channel activator, on changes in cytosolic calcium concentration [( Ca++]i) and tension induced by acetylcholine (ACh; 0.1-10 microM) were examined in swine tracheal smooth muscle. Cromakalim (10 microM) hyperpolarized muscle cells by approximately 18 mV from -58 mV (resting membrane potential) to -76 mV. Cromakalim relaxed muscle contractions evoked by ACh at a concentration of 0.1 microM, but not at higher concentrations. Measurement of [Ca++]i using Fura-2 demonstrated that except at 0.1 microM ACh, cromakalim did not alter peak increases in [Ca++]i. At 0.1 microM ACh, the peak transient was decreased, but not eliminated. Cromakalim reduced steady-state increases in [Ca++]i at ACh less than or equal to 1 microM, but not 10 microM ACh. Tension was similarly affected. These data suggest that ACh-induced increases in steady-state [Ca++]i and tension are inhibited by cromakalim-induced hyperpolarization. The initial ACh-induced transient increase in [Ca++]i is not greatly altered. Cromakalim did not alter the transient peak tension and [Ca++]i relationship. The relationship between steady-state [Ca++]i/tension (EC50 = 321 nM) obtained for control, cromakalim inhibition and after glibenclamide reversal of cromakalim inhibition falls to the left of the peak transient [Ca++]i/tension relationship (EC50 = 587 nM). Thus, the Ca++ sensitivity of the contractile proteins during steady-state stimulation by ACh was increased from that at rest. We conclude that electromechanical coupling is important in ACh-induced contraction at concentrations less than 1 microM. Pharmacomechanical coupling with little or no sensitivity to changes in potential is important at higher ACh concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

PubMed1991

Autonomic stimulation of short circuit current in swine trachea.

Autonomic stimulation of ionic fluxes was studied in tracheal epithelium of weanling swine by measuring short circuit current (ISC) in an Ussing chamber. Basal currents were predominantly secretory since 0.1 mM diphenylamine 2-carboxylic acid (DPC) reduced currents by twice as much as did 10 microM amiloride. Both acetylcholine (ACh) and isoproterenol (ISO) produced a sustained increase in ISC, with the EC50 for ISO being 149 nM and for ACh being 500 nM. In both cases, the induced currents were predominantly secretory since 10 microM amiloride had little effect on either agonist-induced delta ISC whereas DPC inhibited the ISO- and ACh-induced delta ISC's by 30% and 47% respectively. However, responses to ACh and ISO could be distinguished on three grounds. [1] An anatomic gradient existed with smaller responses to agonists being observed towards the distal trachea. The ISO-induced currents showed a steeper gradient than the ACh-induced currents. [2] In single tissue samples, the size of the ISO-induced delta ISC was not correlated with the size of the ACh-induced delta ISC. [3] The magnitude of the delta ISC induced by ACh was unaffected by a concomitant stimulation of ISC by ISO. Thus, ACh-induced secretion was independent of ISO-induced secretion, and likely occurred in different cell types.

Acetylcholine↗

PubMed1991

Intracellular chloride in submucosal gland cells.

The chloride ion concentration within isolated tracheal submucosal gland cells was studied micro-spectrofluorometrically using a fluorescent dye, 6-methoxy-N-(3-sulfopropyl) quinolinium (SPQ), that is quenched by Cl-. Cells from normal weanling swine and from a cystic fibrosis (CF) patient were used. Ion substitution experiments showed that cell fluorescence increased in both cell types when bath Cl- was replaced with the impermeant anion glucuronate. Following a Donnan-type ion substitution that kept the product of the bath K+ and Cl- concentrations constant, reducing bath chloride had little effect on fluorescence for normal cells, but caused a marked increase for CF cells. Thus, K+ and Cl- ions have approximately the same Nernst potential in control submucosal gland cells; in contrast, cells from a CF patient concentrated Cli, resulting in a Cl- Nernst potential that was more positive than the K+ Nernst potential. This finding is consistent with the hypothesis that CF submucosal gland cells have a decreased Cl- permeability.

Animals↗

PubMed1991

Pirenzepine block of ACH-induced mucus secretion in tracheal submucosal gland cells.

Muscarinic stimulation of mucus secretion, as measured by the release of [3H]glycoprotein, was studied in explants from the tracheal epithelium of weanling swine. The mucus glycoprotein secretion was transient, ceasing within the first 10 min of a continuous exposure to 100 microM ACh. Increasing the solution's osmotic pressure did not alter basal mucus glycoprotein secretion. Mucus glycoprotein secretion was inhibited by 2-10 microM PZP, indicating that the M3 muscarinic receptors mediate cholinergic stimulation of mucus production.

Animals↗

PubMed1991

Muscarinic receptors and mucus secretion in swine tracheal epithelium: effects of subacute organophosphate treatment.

This swine trachea study was undertaken to examine the effects of nerve agents on mucus gland cell function. Subacute treatment of swine with soman, sarin, and VX inhibits acetylcholinesterase and leads to down-regulation of muscarinic receptors in tracheal submucosal gland cells. Muscarinic receptor density in isolated cells as determined by [3H]QNB binding was reduced by 60-65% and that measured using [3H]NMS was decreased by 65-73%. Subacute treatment of swine for 7 days with soman and VX caused a small, significant increase in the fraction of receptors with high affinity for carbachol. The decrease in receptor density was accompanied by a decrease in acetylcholine-induced mucus secretion. The decrease in mucus secretion was not due to a decrease in the ability of the cells to produce or release mucus since cross-tolerance did not develop to methoxamine-induced mucus secretion. Therefore, we conclude that in mucus gland cells tolerance development can be linked functionally to muscarinic receptor loss.

Animals↗

PubMed1991

Cocaine and tracheal epithelial function: effects on short circuit current and neurotransmitter receptors.

After snorting crystalline cocaine, the nasal and airway mucosa will be exposed to high concentrations of this drug. We have examined the actions of mucosal and serosal cocaine application on the basal short circuit current (Isc) and the changes in Isc (delta Isc) induced by acetylcholine (ACh) and isoproterenol (ISO) across swine tracheal epithelium. Cocaine displacement of muscarinic receptor and beta adrenoceptor radioligands was also examined. Cocaine at low, less than 1 mM, concentrations induced basal Isc to increase in some tissues by 9 to 10 microA in some preparations it induced only a decrease in basal Isc. The maximal decrease was 15 to 20 microA in these preparations. In all tissues 14 mM cocaine decreased basal Isc. The IC50 for the cocaine inhibition of Isc was 3 +/- 0.9 mM after mucosal application. Mucosal cocaine (3 mM) did not affect the actions of mucosal amiloride, an epithelial sodium channel blocker (IC50 = 0.6 +/- .1 microM, control; 1.5 +/- 0.3 microM, in the presence of 3 mM cocaine) or serosal tetraethylammonium, a potassium channel blocker. However, cocaine altered the response of the tissue to ACh and ISO. Cocaine (14 mM) applied mucosally reduced the maximal delta Isc induced by ACh (serosal) to 34 +/- 14% of control. By contrast, serosal cocaine (3 mM) caused a parallel shift to the right in the concentration-response relationships for ACh- and ISO-induced increases in Isc (EC50 increased by approximately 6.3 and approximately 2.5 times, respectively) but did not reduce the maximal response.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

PubMed1991

Concentration-dependence of acetylcholine-induced changes in calcium and tension in swine trachealis.

Acetylcholine (ACh)-induced increases in intracellular calcium concentration ([Ca++]i) and tension were measured simultaneously in swine tracheal smooth muscle strips loaded with the calcium-sensitive fluorescent dye Fura-2. ACh at concentrations greater than or equal to 3 x 10(-8) M induced concentration-dependent increases in tension which reached a maximum at 10(-4) M. Increases in [Ca++]i occurred at all [ACh] tested (10(-8) to 10(-4) M). After addition of ACh at concentrations greater than 3 x 10(-7) M, [Ca++]i increased rapidly to a concentration dependent-peak then declined to a concentration-independent steady state approximately 250 nM above the resting [Ca++]i of 257 +/- 12 nM. There was a steep relationship (slope factor greater than 3) between the peak tension and the peak [Ca++]i reached at each [ACh]. The rate of decline of [Ca++]i to the steady state at [ACh] greater than 73 x 10(-7) M was well correlated with the peak [Ca++]i reached. We conclude that the peak increase in calcium induced by ACh sets the level of tension to be attained and the rate of decline of the transient increase in [Ca++]i. The steady-state [Ca++]i is sufficient for maintenance of tension.

Acetylcholine↗

PubMed1990

Down-regulation of muscarinic receptors in the striatum of organophosphate-treated swine.

Subacute (daily) administration of diisopropylfluorophosphate (DFP) to male swine (Yorkshire white) resulted in a 97% inhibition of cholinesterase and a decrease of [3H]quinuclidinyl benzilate [( 3H]QNB) binding sites in homogenates of striata by approximately 50% after 14 days. The maximal density of receptors (Bmax) decreased from 2.1 +/- 0.3 to 1.0 +/- 0.2 pmole/mg protein. There was no significant change in the dissociation constant (Kd) for [3H]QNB binding (control: 52.6 +/- 10.7 pM; 7-day: 57 +/- 2.8 pM). Carbachol displacement of [3H]QNB binding yielded data best fit by a two-binding site model. The dissociation constants were KiL = 115 +/- 62 microM (55 +/- 3%) and KiH = 1.8 +/- 0.7 microM (45 +/- 3%), respectively, for the low- and high-affinity states. Seven-Day treatment with DFP reduced the percentage of high-affinity receptors to 22 +/- 8.6%, but affected neither the low- nor the high-affinity Kd (100 +/- 20 and 2 +/- 0.6 microM). With the addition of Mg2+, striatal homogenates had low- and high-affinity receptors in the proportion of approximately 1 to 1. In the presence of Gpp(NH)p + Mg2+ the ratio of high- to low-affinity receptors was 3:1 in homogenates of control tissue (to 26 +/- 5%). This treatment had no effect on this ratio in homogenates of tissue from 7-day DFP-treated swine (3:1) since it was already 3:1. Pirenzepine displacement of [3H]QNB binding was best described by a two-binding site model, with Ki values of 38 +/- 14 and 201 +/- 78 nM, which represent 74 and 26% of the binding sites, respectively. The high affinity Kd value was unchanged following 7 days of DFP treatment (24 +/- 5 nM). There appears to be little change in the displacement curves for pirenzepine inhibition of [3H]QNB binding. This suggests that about 75% of the receptors are of the M1 subtype. Thus, subacute administration of DFP causes not only a decrease in the number of receptors, but also a change in the proportion of agonist affinity states which is related to the interaction of the guanine nucleotide binding protein and the muscarinic receptor.

Acetylcholinesterase↗

PubMed1990

Communication between the PC and laboratory instruments.

In this paper we describe communications between a personal computer and any instrument with a serial port for the purpose of collecting data or controlling equipment and a program for performing this task. From a user-defined file, the software reads the communication parameters: baud rate, data bits, stop bits and parity. The communication protocol is also read: echo, acknowledge and end-of-transmission characters. The software can be used to determine the protocol, to check the integrity of the communications, and can form the basis of a program for more specific applications.

Equipment and Supplies↗

PubMed1989

An integrating averager of microspectrofluorometric signals.

Optical signals are used to measure a variety of cell events; in particular, ratiometric fluorescent dyes offer diverse applications, but bring with them the difficult analytical problem of obtaining optical signals at rapidly alternating wavelengths. An improved method for using a computer to record these optical signals is described, and an example record of a Ca+2 transient given. A second technical problem is that ultraviolet (UV) light is required to excite many fluorescent dyes; many common lenses cannot transmit UV light. This article describes the tests needed to measure and optimize the optical path taken by light through a microscope and provides examples with sample lenses.

Animals↗

PubMed1988

Changes in affinity states during down-regulation of muscarinic receptors in tracheal smooth muscle of organophosphate-treated swine.

1. Subacute (daily) treatment of male swine with the organophosphate acetylcholinesterase inhibitor diisopropylfluorophosphate (DFP) resulted in tolerance to the effects of DFP within 5-6 days. 2. Subacute administration of DFP resulted in a 98% inhibition of tissue cholinesterase after 7 days and in a decrease of [3H]quinuclidinyl benzilate [( 3H]QNB) binding sites in homogenates of tracheal smooth muscle by 77%. The maximal density of receptors (Bmax) decreased from 1.8 +/- 0.4 to 0.5 +/- 0.1 pmole mg-1 protein. There was no significant change in the dissociation constant (Kd) for [3H]QNB binding. 3. Pirenzepine displacement of [3H]QNB binding was best described by a single binding site model, with a Ki of 230 +/- 40 nM. This value was unchanged following seven days of DFP treatment (250 +/- 30 nM). The low affinity for this M1 antagonist suggests that there is predominantly a single population of [3H]QNB binding sites of the M2 subtype in tracheal smooth muscle. 4. Carbachol displacement of [3H]QNB binding yielded data best fit by a two-binding site model. The dissociation constants were KiL = 210 +/- 60 microM (61 +/- 1%) and KiH = 1.2 +/- 0.4 microM (39 +/- 1%) respectively (n = 7) for the low and high affinity states. Seven-day treatment with DFP reduced the percent of high affinity receptors to 25 +/- 4%. 5. Addition of Mg++ to the incubation medium prevented this shift in the proportion of low and high affinity receptors. Gpp(NH)p and Mg++ together decreased the proportion of the high affinity receptors when added to the incubation medium in control tissue (to 25%), but not tissue from 7-day DFP-treated swine. NEM increased the proportion of muscarinic receptors in the high affinity state both for controls and for the DFP-treated swine, in both cases yielding receptors with identical binding properties. 6. Thus, subacute administration of DFP causes not only a decrease in the number of receptors, but also a change in the affinity of the receptors for agonists which is related to the interaction of the guanine nucleotide binding protein and the muscarinic receptor.

Animals↗

PubMed1988

Contractile responses of tracheal smooth muscle in organophosphate-treated swine: 1. Agonist changes.

1. Male weanling swine were injected daily for up to 14 days with the organophosphate cholinesterase inhibitors, diisopropylfluorophosphate (DFP) or sarin. The clinical signs of poisoning disappeared or were attenuated by 7 days after starting the DFP treatment, indicating the development of tolerance to DFP toxicity. 2. A significant decrease in acetylcholinesterase activity (85-98%) occurred over the course of this treatment followed by a decrease in the maximal density (Bmax) of [3H] quinuclidinyl benzilate ([3H]QNB) and [3H] N-methylscopolamine ([3H]-NMS) binding sites in isolated cells. The affinity of the muscarinic receptors (KD) for [3H]QNB and [3H]NMS binding, however, remained unaffected. 3. Dose-response curves for ACh-induced increase in isometric tension of tracheal smooth muscle (TSM) showed a leftward shift from control, 2 h after DFP injection. Twenty-four hours after the last DFP treatment, for animals receiving 1 or up to 14 daily injections of DFP, all the dose-response curves were shifted to the left to approximately the same ACh sensitivity when compared with that for control tissue. 4. In vitro treatment of the muscle with 10(-4) M DFP shifted the dose-response curves leftward, in both control and injected animals, and rendered the muscles from control, 1- and 3-day injected animals sensitive to ACh concentrations as low as 10(-10) M. Sensitivity to 10(-10) M ACh was eliminated by carefully cleaning the smooth muscle of adherent connective tissue containing nerves and ganglia and after subacute treatment of swine for 7 days with DFP. DFP-induced spontaneous contractions were also eliminated by careful cleaning. 5. Subacute DFP treatment caused a small leftward shift in the dose-response curve for bethanechol at 2 h and a rightward shift at 1,3 and 7 days, compared to controls. 6. Dose-response curves for K+ were shifted to the right after 1 and 3 days of DFP treatment, but shifted back towards the control after 7 days of treatment. The muscle cells were hyperpolarized by approximately 5 mV after 7 days of DFP or sarin injections. The membrane potential was slightly more sensitive to changes in K+ concentration after 7 days of sarin injection. 7. Subacute treatment of swine with organophosphates modifies the response of neural elements in swine TSM to ACh. Chronic cholinesterase inhibition causes a reduction in the sensitivity of the neural elements to ACh. The decrease in muscarinic receptor density which occurs with chronic cholinesterase inhibition is not sufficient to explain tolerance to organophosphates since TSM maintains an almost normal responsiveness to ACh.

Acetylcholine↗

PubMed1988

Contractile responses of tracheal smooth muscle in organophosphate-treated swine: 2. Effects of antagonists.

1. Swine tracheal smooth muscles (TSM) developed spontaneous contractions following the acute administration of DFP in vivo and/or in vitro which could be blocked pharmacologically using atropine (2 x 10(-7) M), pirenzepine (3 x 10(-7) M), or hemicholinium 3 (HC3, 5 x 10(-6) M). 2. Treatment of TSM in vitro with DFP caused them to become responsive to ACh concentrations as low as 10(-10) M. 3. Atropine and pirenzepine (at 2 and 3 x 10(-7) M respectively) increased the EC50 concentrations for ACh approximately 300- and 8-fold respectively. The shifts caused by atropine and pirenzepine in the dose-response curves for ACh were not parallel after in vitro treatment of the muscle with DFP. By contrast, the shifts in the dose-response curves were parallel when muscles from swine injected for 7 days with DFP were used. 4. HC3 had no effect on the control dose-response curve for ACh, but steepened the dose-response curve after in vitro treatment of the muscle with DFP. The ACh dose-response curve obtained in the presence of HC3 and DFP was identical to that obtained using muscles from swine treated for 7 days with DFP. 5. McN-A-343, a partial agonist at muscarinic receptors, also induced contraction although the maximal tension induced was 56% of the maximal tension obtained using ACh. In vitro treatment of the muscle with DFP caused a leftward shift in the dose-response curve for McN-A-343. The muscles from animals treated for 7 days with DFP did not respond to McN-A-343 at doses up to 10(-3) M. 6. McN-A-343 competition for [3H]QNB binding suggested that the loss of the contractile response can be correlated with the loss of a high affinity site for McN-A-343 from the muscle. 7. We conclude that tolerance to DFP with subacute treatment results in part from the reduction in sensitivity of neural elements associated with swine tracheal smooth muscle to ACh. In addition the response to the partial agonist McN-A-343 is lost after subacute DFP treatment.

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy↗