Cortisol and cortisone levels in umbilical cord plasma and maternal plasma of normal pregnancies

A method for assaying cortisol and cortisone using chromatography on either paper or Sephadex LH-20 columns for isolation, followed by competitive protein binding, has been applied to umbilical cord and maternal plasma samples. In mixed cord plasma the mean cortisol concentration was 6.0 ± 0.8 μg/100 ml (n = 9) and the mean cortisone concentration was 13.5 ± 2.9 μg/100 ml (n = 9). In cord arterial plasma the mean cortisol concentration was 6.3 ± 2.9 μg/100 ml (n = 6) and the mean cortisone level was 10.1 ± 2.5 μg/100 ml (n = 6). For cord venous plasma, the mean level of cortisol was 5.6 ± 1.5 μg/100 ml (n = 6) and of cortisone was 13.5 ± 2.4 μg/100 ml (n = 6). Maternal plasma gave a mean value of cortisol of 42.3 ± 4.5 μg/100 ml (n = 6) and of cortisone of 6.2 ± 0.9 μg/100 ml. The results of this study suggest that the fetus at term-gestation produces cortisol. The significance of this production compared with placental transfer of maternal cortisol into the fetal circulation however is uncertain.     

Mechanism of spasmolytic activity of a fraction of Sarcostemma brevistigma Wight

The effect of chloroform soluble fraction (F-A) of twigs of Sarcostemma brevistigma on contractions induced by KCl, histamine, and acetylcholine in the isolated guinea pig ileum and taenia coli smooth muscles has been evaluated. F-A (19.5 microg/ml) significantly inhibited the contraction induced by 40 mM KCl to the extent of 87.6% in the isolated guinea pig ileum. In the isolated guinea pig ileum, F-A (64.3 and 59.2 microg/ml) significantly inhibited the contractions induced by acetylcholine and histamine to the extent of 85 and 83% respectively. In the isolated guinea pig taenia coli, F-A (65.2 microg/ml) significantly inhibited the contraction induced by 40 mM KCl to the extent of 96.0%. The inhibitory effect of F-A (40 microg/ml) on the isolated guinea pig taenia coli was reduced by Bay K 8644 (10(-6) M) to the extent of 61.6 from 73.6%. These results suggest that the F-A may exhibit smooth muscle relaxant activity by blocking the Ca2+ channels.     

Antigen-induced elevation of immunoreactive endothelin-1 (ET-1) levels in ovalbumin-sensitized guinea pig airway tissue

Changes in the immunoreactive ET-1 levels during the anaphylactic reaction of airway tissue from ovalbumin-sensitized guinea pigs were investigated. ET-1-immunoreactivity (ET-IR) was detected in the epithelial and smooth muscle layers of tracheal sections from normal guinea pigs and it was enhanced slightly by phosphoramidon (1 μM) treatment. The ET-IR level of the epithelial layer of ovalbumin-treated tissue from actively sensitized animals was slightly higher than that from normal animals, but it was enhanced markedly by phosphoramidon (1 μM) treatment. Furthermore, the mean ET-IR level of homogenates of antigen-treated tracheal tissues from sensitized guinea pigs (22.8±1.55 fmol mg⁻¹ protein, n=5) was significantly higher than the corresponding normal level (12.3±1.21 fmol mg⁻¹ protein, n=5). These results suggest that increased epithelial airway ET-1 levels contribute to the anaphylactic reaction of guinea pig airways.     

Vasoactive intestinal polypeptide: increased tone, enhancement of acetylcholine release, and stimulation of adenylate cyclase in intestinal smooth muscle

Vasoactive intestinal polypeptide (VIP) was examined $\text{in vitro}$ for effects on tone and neuronal release mechanisms in intestinal smooth muscle since this is a site of high peptide concentration. VIP contracted the guinea pig ileum and rabbit jejunum in concentrations ranging from 10^?9 to 10^?7 M. Increased tone in the guinea pig ileum was partially antagonized by the anticholinergic agent, atropine (4.38 × 10^?6 M) suggesting that one component of the contractile response was due to the indirect release of acetylcholine. The H₁ receptor antagonist, pyrilamine, did not alter the increased tone produced by VIP indicating that histamine release did not contribute to the ileal contractile response and that VIP exerted a selective effect to enhance neuronal release of acetylcholine. The ability of VIP to modulate acetylcholine release was confirmed in field stimulated ileal preparations where VIP increased the force developed to endogenously released acetylcholine without altering the direct response to acetylcholine. In rabbit jejunum and ileal smooth muscle, VIP related cyclic AMP levels. However, inhibition of phosphodiesterase with papaverine did not potentiate either the VIP-induced ileal contraction or enhancement of the field stimulated response. This raises the possibility that increases in intestinal cyclic AMP may be involved more in VIP-induced alterations in ion transport or secretory phenomenon than in intestinal motility. These studies describing the ability of VIP to modulate acetylcholine release and to increase ileal tone are consistent with the proposed role of VIP in intestinal patholgies involving excessive mucous secretion and motility.     

The production and secretion of cortisol by baboon neonates.

The metabolic clearance rate (MCR), transfer constants (p), production (PR) and secretion (SR) rates of cortisol (F) andrortisone (E) were determined by the continuous infusion of {1,2³H}F and {4-¹⁴C}E into 5 neonates delivered prior to the parturition by cesarean section (164–179 days; term = 184 days) and into 5 newborns delivered spontaneously per vagina at term (166 – 187 days). In spontaneously delivered animals, MCR-E (X ± SE, 34.3 ± 7.0 1/day/kg was greater (P < 0.001) than MCR-F (14.9 ± 1.5 1/day/kg), pF to E (59.7 ± 8.9%) exceeded (P < 0.001) pE to F (17.8 ± 3.0%) and the percentage of F bound to serum proteins other than albumin (57.5 ± 6.2) was greater (P < 0.001) than that of E (27.0 ± 10.3) Although the serum E level (25.6 ± 3.6 μg/100 ml) was similar to that of F (33.5 ± 8.0 μg/100 ml), the PR-E (6.4 ± 1.3 μ/min/kg) was greater (P < 0.001) than PR-F (3.3 ± 0.5 μ/min/kg). Approximately eighty-five percent of the E and 65% of the F produced orginated by secretion.In animals delivered by cesarean section, the serum F concentration (32.4 ± 6.7 μ/100ml), pE to F (13.4 ± 2.8%) pF to E (80.0 ± 12.2%) PR-E (4.5 ± 0.2 μ/min/kg) and SR-E (3.9 ± 0.3 μ/min/kg) were not different from values for spontaneously delivered animals. Serum E levels (35.9 ± 1.6 μ/100 ml) were higher but MCR-F (6.7 ± 0.6 1/day/kg) and MCR-E (18.2 ± 0.41/day/kg) lower in neonates delivered by cesarean section. Serum Cortisol binding capacity (μg F bound/100 ml) was greater (P < 0.025) in neonates delivered by cesarean section (23.6 ± 2.6) than in spontaneously delivered animals (14.4 ± 2.0). As a result of these changes in F and E dynamics, PR-F (1.4 ± 0.3 μ/min/kg) and SR-F (0.9 ± 0.2 μ/min/kg) in neonates delivered by cesarean section were lower (P< 0.01) than corresponding values in spontaneously delivered newborns.It is concluded that the greater F secretion in animals delivered spontaneously than those delivered by cesarean section probably results from increased fetal adrenal 3β-hydroxysteroid dehydrogenase-isomerase activity, which as previously reported, occurs in late gestation in this species.     

Biological activity and anti-prostaglandin effects of prostaglandin analogue, ent-11-epi-15-epi PGE2 methyl ester

The biological activity and anti-prostaglandin property of the prostaglandin analogue, ent-11-epi-15-epi PGE2 methyl ester, were studied on isolated guinea pig ileum. Ent-11-epi-15-epi PGE2 methyl ester contracted guinea pig ileum and produced a concentration-response curve parallel to that of PGE2. However, the former exhibited a lower maximal effect than PGE2. At concentrations greater than 10(-6)M, ent-11-epi-15-epi PGE2 methyl ester selectively antagonized contractile actions of PGE2 and PGE2 alpha without affecting contractions induced by acetylcholine. These observations suggest that the PG analogue acted like a competitive antagonist to PGE2 and PGF2 alpha on guinea pig ileum in vitro.     

The effect of calcium antagonists on contractions of the sensitized and normal guinea pig ileum

The purpose of this study was to learn wether a number of Ca²⁺ antagonists were effective in reducing contractile response of the isolated ileum of the sensitized and normal guinea pig. Contractions of the normal ileum in response to LTD₄, acetylcholine, histamine, and potassium chloride were obtained before and after verapamil, diltiazen and papaverine. Ovalbumin-induced contractions of the ovalbumin-sensitized ileum were obtained in the presence of the three Ca²⁺ antagonists. In the normal ileum, all the Ca²⁺ antagonists were highly effective in diminishing the contractile responses to LTD₄, acetylcholine, histamine and potassium chloride. In the sensitized ileum, ovalbumin-evoked contractions, with subsequent release of a potent contractile mediator (presumably SRS-A), were Ca²⁺-dependent since verapamil, diltiazem and papaverine caused a concentration-related reduction of contractions. Thus, the influx of extracellular Ca²⁺ plays a key role in the contractile responses of the normal and sensitized guinea pig ileum when stimulated by various potent agonists acting on specific receptors or on the cell membrane.     

Prejunctional alpha 2 adrenoceptors inhibit contraction of tracheal smooth muscle by inhibiting cholinergic neurotransmission

Ring preparations obtained from the guinea pig trachea contracted on short trains of electrical field stimulation. These contractions were mediated by activation of cholinergic nerves since they were abolished by atropine or tetrodotoxin. In the presence of beta blocking drugs noradrenaline and adrenaline dose-dependently inhibited contractions induced by field stimulation. By contrast, contractions on exogenous acetylcholine were left completely unaffected. It is concluded that the adrenergic agonists inhibited cholinergic neurotransmission by a prejunctional action. In order to characterize the noradrenaline receptor the effects of alpha₁ and alpha₂ blockers were evaluated using the Schild plot. For comparison experiments were also conducted on the guinea pig aorta and electrically stimulated guinea pig ileum. The results indicate that in guinea pig trachea and ileum noradrenaline inhibits cholinergic neurotransmission by acting on prejunctional alpha₂ receptors whereas in guinea pig aorta it induces contraction by stimulating alpha₁ receptors.     

Immediate x-radiation induced contractions of the isolated guinea pig terminal ileum: The localization of the effect by drugs

Summary Tone and motility of the isolated guinea pig ileum were increased by irradiation with a dose of 10 krd. The maximal effect corresponds to that induced by 0.001 µg/ml acetylcholine or 0.3 µg/ml nicotine. The pharmacological analysis of this effect performed with acetylcholine and nicotine and several blocking agents including hexamethonium, atropine, tetrodotoxin, diphenhydramine, and verapamil suggests that radiation acts on the postganglionic parasympathetic neuron and the neuromuscular synapse. The mechanism of radiation is likely to consist of both an increased release of acetylcholine from the postganglionic neuron and a sensibilization of the cholinergic receptor site at the smooth muscle cell. The latter effect is thought to result from an increased contractile action induced by acetylcholine or nicotine in the irradiated ileal smooth muscle.     

Alpha-adrenergic receptor blocking effect of Cleistanthus collinus (Roxb.) Benth. and Hook f. leaf extract on guinea pig isolated smooth muscle preparations

Aqueous extract of C. collinus leaves inhibited norepinephrine induced contraction in guinea pig vas deferens and aortic strip in a dose-dependent manner. Inhibition of acetylcholine induced contraction in ileum was dose independent. C. collinus extract per se had no effect on isolated guinea pig vas deferens and aortic strip, but inhibited norepinephrine induced contraction in a dose-dependent manner probably by its antagonist action on alpha-adrenergic receptor. It had inconsistent effect on guinea pig ileum in vitro preparation.     

Lithium does not synergize the peripheral action of cholinomimetics as seen in the central nervous system

Lithium is known to synergize the action of cholinomimetics in the CNS such that pilocarpine induces seizures in low concentration (1/13th of per se dose) in rats. The present study was undertaken to see if lithium priming also enhances the peripheral effects of acetylcholine and pilocarpine i.e. change in blood pressure in rats and contractions of the isolated guinea pig ileum. In anaesthetized rats the blood pressure was recorded from cannulated carotid artery connected through the pressure transducer to Coulbourn polygraph. The blood pressure response of pilocarpine was not different either in magnitude or in duration when administered 1, 2 and 4 h after lithium chloride (3 meq/kg) pretreatment as compared to the control. Similarly acetylcholine effect remained unchanged after lithium chloride priming. In the isolated guinea pig ileum experiments, ileum was incubated for 1 h in different concentrations of lithium chloride and effect on acetylcholine induced contractions were observed. Lithium in concentration of 2.8 x 10(-3) M had no effect on acetylcholine induced contractions while incubation with higher concentrations of 1.4 x 10(-2) M and 2.8 x 10(-2) M significant inhibition of acetylcholine contractions were observed. At this concentration, histamine induced contractions were also inhibited. The results indicate that lithium does not synergize the action of cholinomimetics in the periphery as that seen in the CNS. The inhibition of acetylcholine and histamine induced contractions in guinea pig ileum at high concentration of lithium seems to be non-specific effect.     

Inhibition of d(-)-3-hydroxybutyrate dehydrogenase by malonate analoges

(1) d(-)-3-Hydroxybutyrate dehydrogenase activity from guinea pig, rat, and bovine heart and from guinea pig liver is inhibited by malonate and tartronate, and more potently by the analogs methylmalonate, bromomalonate, chloromalonate, and mesoxalate. Little or no inhibitory effect was found for aminomalonate, ethylmalonate, dimethylmalonate, succinate, glutarate, oxaloacetate, malate, propionate, pyruvate, d- and l-lactate, n-butyrate, isobutyrate, and cyclopropanecarboxylate. (2) In initial velocity kinetics at pH 8.1 with a soluble enzyme preparation from bovine heart, the inhibition by the active malonate derivatives is competitive with respect to 3-hydroxybutyrate and uncompetitive with respect to acetoacetate, NAD⁺ or NADH. With d-3-hydroxybutyrate as the variable reactant (K_{m app} = 0.26 mM) the inhibition constant of methylmalonate (K_is) was 0.09 mm. (3) The rate of utilization of d-3-hydroxybutyrate (78 μm) by coupled rat heart mitochondria in the presence of ADP was inhibited 50% by 150 μm methylmalonate. (4) With coupled guinea pig liver mitochondria oxidizing n-octanoate in the absence of added ADP, methylmalonate (1–3 mm) depressed 3-hydroxybutyrate formation substantially more than total ketone production. However, the intramitochondrial NADH (or NADPH) levels were unchanged by the addition of methylmalonate, indicating that the changes in ratios of accumulated 3-hydroxybutyrate and acetoacetate were caused by direct inhibition of 3-hydroxybutyrate dehydrogenase. Methylmalonate had the same effect on 3-hydroxybutyrate/acetoacetate ratios and ketone body formation with pyruvate or acetate as the source of acetyl groups. Similar results were obtained with malonate (10 mm) although the inhibition of total ketone formation from octanoate was more severe.     

Inhibition of dopamine receptors in the stomach: An explanation of the gastrokinetic properties of domperidone

We investigated the effects of dopamine and secrenin on the motility of the isolated guinea pig stomach. Changes in intragastric volume were continously recorded as a measure of gastric motility. When 0.25 μg/ml dopamine was given intra-arterially (i.a.) via the coeliac axis, the stomach relaxed, its spontaneous activity decreased, and its content augmented. This effect of dopamine could be selectively inhibited by the dopamine antagonists haloperidol (0.16 μg/ml) and domperidone (0.16 μg/ml); the latter is effective in the treatment of gastro-intestinal dysfunction. These observations demonstrate the presence of dopaminergic receptors in the guinea pig stomach. Similar conclusions can be reached for the rat stomach from ³H-domperidone displacement studies. Secretin (6.7 mU/ml i.a.) inhibited the vagally induced emptying of the stomach. This effect is reversed by domperidone (0.04 μg/ml). These results strongly suggest that dopamine or/and secretin are involved in the local feedback control of gastric motility; the action of secretin may be direct or indirect by release of dopamine. The local interference of domperidone with endogenous secretin and dopamine in the stomach may explain its gastrokinetic properties.     

Effect of antihistaminics on bradykinin action]

On isolated ileum of the guinea pig the antihistamine drugs dimedrol, diprazin (pipolphen), tavegil and suprastin diminished spasmogenic effects of bradykinin and an enhancement of microvascular permeability induced by this polypeptid. The antihistamine drugs display a nonspecific antibradykinin activity. No relationship between the chemical structure and pronounced antibradykinin action of the antihistamines studied was revealed.     

The possible role of intracellular receptors in the expression of narcotic antagonist precipitated abstinence

The antagonist potencies of methylnaloxone and naloxone were measured for morphine in the isolated guinea pig ileum by the pA₂ method and a potency ratio determined. A second potency ratio was determined by comparing the concentration of each antagonist required to produce abstinence-like contractures of ileal tissues from morphine-treated guinea pigs. If the latter response required penetration of the cell membrane lipid bilayer, one would expect methylnaloxone, a quaternary amine, to be significantly less potent than naloxone. Comparison of potency ratios for methylnaloxone and naloxone did not support this hypothesis.     

Isolation and characterization of smooth muscle cell membranes

A method for the isolation of guinea pig ileum smooth muscle cell membranes is described. The plasma membrane fraction possessed a (Na⁺, K⁺)-ATPase which was inhibitied by ouabain. The Mg²⁺-dependent ATPase of the membrane fraction was stimulated by 1 μM Ca²⁺. A basal ATPase, not dependent on Mg²⁺, was directly stimulated by Ca²⁺ in the range of 1 μM to 1 mM.The isolated membranes contracted in response to the following substances: ATP, angiotensin II and some of its analogs, bradykinin, acetylcholine and histamine. The contractility was inhibited by ouabain and chlorambucil-angiotensin II, but not by cytochalasin B. No contraction was produced by AMP, angiotensin I and adrenaline.     

The validity of the cholesterol nucleation assay

The validity of the cholesterol nucleation assay rests on the assumption that all cholesterol crystals are removed at the start of the assay so that de novo formation of crystals can be studied. In this paper we have tested the validity of this assumption. Cholesterol crystals were added to supersaturated model bile. Subsequently the mixtures were either filtered over a 0.22 μm filter or centrifuged at 37°C for 2 h at 100 000 × g. After ultracentrifugation the isotropic interphase was collected. Using polarized light microscopy no crystals could be visualized in this fraction. However, the nucleation time of the isotropic interphase decreased from 6.8 ± 1.1 days to 1.8 ± 0.2 days (mean ± S.E., P < 0.01, n = 5) when 10–100 μg/ml crystals were added prior to centrifugation. Similar results were observed when instead of centrifugation the mixtures containing crystals were filtered. After filtration over a 0.22 μm filter no crystals could be detected in the filtrate. Yet the nucleation time of the filtrate decreased from 6.4 ± 0.7 days to 3.1 ± 0.5 days (mean ± S.E.) when 10 μg/ml cholesterol crystals were added before filtration (n = 10, P < 0.01). Since no cholesterol crystals could be detected at the start of the assay the reduction in nucleation time must have been brought about by cholesterol microcrystals that passed through the filter. Supplementation of cholesterol crystals to model bile did not accelerate the nucleation time when the samples were passed over a 0.02 μm filter, indicating that the size of the microcrystals was larger than 20 nm. The effect of addition of cholesterol crystals prior to filtration over a 0.22 μm filter was also tested in the crystal growth assay recently developed by Busch et al. ((1990) J. Lipid Res. 31, 1903–1909). Addition of crystals had only a minor effect on the assay. In conclusion, the reduced nucleation time of biles from gallstone patients is probably not only due to the presence of promoting or the absence of inhibiting proteins, but can be caused by the presence of small cholesterol crystals in these biles.     

L-648,051, sodium 4-[3-(4-acetyl-3-hydroxy-2-propylphenoxy)- propylsulfonyl]-gamma-oxo-benzenebutanoate: a leukotriene D4 receptor antagonist

L-648,051, sodium 4-[3-(4-acetyl-3-hydroxy-2-propylphenoxy) propylsulfonyl]-gamma-oxo-benzenebutanoate is a selective and competitive inhibitor of [3H]leukotriene D4 (KB value of 4.0 microM) and to a lesser extent [3H]leukotriene C4 (Ki value of 36.7 microM) binding in guinea pig lung homogenates. Functionally, it selectively antagonized contractions of guinea pig trachea induced by leukotrienes C4, D4, E4, and F4 in concentrations that did not antagonize contractions induced by acetylcholine, histamine, serotonin, prostaglandin F2 alpha, or U-44069 (endoperoxide analogue). Schild plot analysis indicated that L-648,051 competitively antagonized contractions of guinea pig ileum induced by leukotriene D4 (pA2 7.7) and contractions of trachea induced by leukotrienes D4, E4, and F4 (pA2 7.3, 7.4, and 7.5, respectively). Contractions of guinea pig trachea induced by leukotriene C4 were inhibited in a noncompetitive fashion (Schild plot slope, 0.45). Developed contractions of trachea induced by the leukotrienes were rapidly reversed by L-648,051 greater than FPL-55712 greater than L-649,923. Intravenous L-648,051 selectively blocked bronchoconstriction induced in anaesthetized guinea pigs by intravenous leukotrienes C4, D4, and E4 but not that induced by arachidonic acid, serotonin, U-44069, or acetylcholine. The compound displayed poor activity following intraduodenal administration. The profile of activity for L-648,051 indicates that it may be a useful topical agent for studying the role of leukotrienes in diseases such as bronchial asthma.     

Pharmacology of L-660,711 (MK-571): a novel potent and selective leukotriene D4 receptor antagonist

L-660,711 (3-(3-(2-(7-chloro-2-quinolinyl)ethenyl)phenyl) ((3-dimethyl amino-3-oxo propyl)thio)methyl)thio)propanoic acid is a potent and selective competitive inhibitor of [3H]leukotriene D4 binding in guinea pig (Ki value, 0.22 nM) and human (Ki value, 2.1 nM) lung membranes but is essentially inactive versus [3H]leukotriene C4 binding (IC50 value in guinea pig lung, 23 microM). Functionally it competitively antagonized contractions of guinea pig trachea and ileum induced by leukotriene (LT) D4 (respective pA2 values, 9.4 and 10.5) and LTE4 (respective pA2 values, 9.1 and 10.4) and contractions of human trachea induced by LTD4 (pA2 value, 8.5). L-660,711 (5.8 x 10(-8)M) antagonized contractions of guinea pig trachea induced by LTC4 in the absence (dose ratio = 28) but not in the presence of 45 mM L-serine borate (dose ratio less than 2). L-660,711 (1.9 x 10(-5)M) did not block contractions of guinea pig trachea induced by histamine, acetylcholine, 5-hydroxytryptamine, PGF2 alpha, U-44069, or PGD2. In the presence of atropine, mepyramine, and indomethacin, L-660,711 (1.9 x 10(-5)M) inhibited a small component of the response to antigen on guinea pig trachea but completely blocked anti-IgE-induced contractions of human trachea. L-660,711 (i.v.) antagonized bronchoconstriction induced in anesthetized guinea pigs by i.v. LTC4, LTD4, and LTE4 but did not block bronchoconstriction to arachidonic acid, U-44069, 5-hydroxytryptamine, histamine, or acetylcholine. Intraduodenal L-660,711 antagonized LTD4 (0.2-12.8 micrograms/kg)-induced bronchoconstriction in guinea pigs, and p.o. L-660,711 blocked LTD4- and Ascaris-induced bronchoconstriction in conscious squirrel monkeys and ovalbumin-induced bronchoconstriction in conscious sensitized rats treated with methysergide (3 micrograms/kg). The pharmacological profile of L-660,711 indicates that it is a potent, selective, orally active leukotriene receptor antagonist which is well suited to determine the role played by LTD4 and LTE4 in asthma and other pathophysiologic conditions.     

Identification of 5HT2-Receptors on Longitudinal Muscle of the Guinea Pig Ileum

Abstract

In binding experiments with the radioligands [³H]Ketanserin (HKet) and [¹²⁵I]LSD (ILSD) 21 compounds were investigated using rat brain cortex membranes. The pK_D-values of the compounds were virtually independent of the radioligand used and their rank order was consistent with classification of the binding sites as being of the 5-HT₂-type. In contrast, in the longitudinal muscle of the guinea pig ileum in the presence of 0.3 μM cinanserin, ILSD labelled sites which were quite different to those in the cortex. In a functional test antagonism of the 5HT induced contraction of the guinea-pig ileum was measured in the presence of 1 μM atropine. The pharmacological inhibition constants (IC₅₀-values) of 8 compounds correlated well with the dissociation constants for HKet binding in the cortex and did not correlate with the data from ILSD binding in the guinea pig ileum. It is concluded that the ileum contains postjunctional 5HT₂-receptors which mediate contraction. The nature of the ILSD binding sites in the ileum remains to be elucidated.