Muscarinic Receptor Blocker Research Articles

Operant Behavior in Conditions of Activation and Blockade of Neostriatal Muscarinic Receptors

Chronic experiments were performed on four dogs using a model of an operant defensive reflex associated with maintaining a flexion posture to study the effects of bilateral intraneostriatal microinjection of the non-selective muscarinic receptor agonist carbachol, the selective D2 dopamine receptor blocker raclopride, and the selective M1 muscarinic receptor blocker pirenzipine on the performance of the operant defensive reflex and differentiation of signals. The results show that microinjection of carbachol induced increases in the tonic component and inhibition of the phasic component of the reflex, an ordering rearrangement of the posture, and increases in the amplitudes of its components. Raclopride microinjection gave similar but less marked results. The greatest effects with both substances were seen using differential stimuli. There were sharp increases in the process of differentiation of sound signals. Pirenzipine microinjections gave the opposite result. These data are assessed on the basis of concepts of the existence of two efferent outputs from the neostriatum with opposite effects on their targets and the roles of muscarinic and dopamine receptors in triggering and blocking these effects.

Contractile effect of acetylcholine on isolated ampullar segment of fallopian tubes

The Fallopian tubes are scarcely innervated with cholinergic nerve fibers. Acetylcholine released from these nerves contracts smooth muscles of the tubes. The aim of our study was to investigate the effect of acetylcholine on the Fallopian tubes using selective antagonists in different hormonal settings. We have investigated effects of acetylcholine on isolated ampulla of Fallopian tubes taken from 83 patients during abdominal hysterectomy with adnexectomy. Twenty-eight patients were in follicular, 36 were in luteal phase of menstrual cycle, and 19 patients were in menopause. Selective and non-selective muscarinic and nicotinic receptor blockers were used for investigation of the effects. Acetylcholine (from 1.8 to 658.6 μM/l) produced concentration-dependent tonic contraction of ampulla taken from the patients in follicular phase, luteal phase and menopause. The nicotinic receptor blocker mecamylamine (6.5 μM/l) and local anesthetic lidocaine (230.8 μM/l) did not affect the effect of acetylcholine. While M 1- and M 2-selective muscarinic receptor blockers pirenzepine (1.6 μM/l) and methoctramine (0.9 μM/l) did not show specific effect, atropine (0.01 μM/l) and selective M 3-receptor blocker p-fluoro-hexahydro-sila-difenidol ( pFHHSiD, 0.2 μM/l) effectively blocked contractions caused by acetylcholine (maximal p A 2 values 9.74 and 7.54, respectively). The affinity of pFHHSiD for muscarinic receptors was highest in the follicular phase. The results of our study suggest existence of functional M 3 muscarinic receptors in ampulla of Fallopian tubes, located on the smooth muscle cells.

The evaluation of the effects of renal failure on erectile dysfunction in a rabbit model of chronic renal failure.

To determine whether chronic renal failure (CRF) reduces nitrergic relaxant responses in a rabbit model. Ten rabbits underwent surgery to induce uraemia (CRF rabbits) and a further 10 a sham operation (controls). Corpus cavernosal tissue was prepared and used in organ-chamber experiments, with relaxation assessed against a background of pre-contraction with phenylephrine. At the plateau of contraction, relaxation responses to cumulative concentrations of carbachol or sodium nitroprusside (SNP), to test endothelium-dependent and -independent relaxations, respectively, were assessed. Before electrical-field stimulation (EFS), the tissue was treated with an adrenergic nerve blocker and a muscarinic receptor blocker to eliminate the adrenergic and cholinergic components, and to determine the relaxation responses to the stimulation of nonadrenergic, noncholinergic (NANC) nerves. The relaxation responses in corporal strips obtained from CRF rabbits were compared with those from controls. When tissues were contracted with KCl, tensions were similar in all groups. The impairment in concentration-dependent relaxation with carbachol was significant in CRF rabbits, but SNP- and papaverine-induced concentration-dependent relaxation responses were no different among the groups. EFS-induced frequency-dependent relaxations were significantly lower in CRF rabbits than in controls. CRF inhibits the NANC-mediated relaxation of rabbit corpus cavernosum smooth muscle. Changes in NANC-mediated and carbachol-induced (endothelium-dependent) relaxation of corporal smooth muscle in the rabbit are probably caused by uraemia and subsequently, hyperthyroidism, hyperparathyroidism or low testosterone levels in CRF. These results also suggest that if vasoactive agents are to be used for treating erectile dysfunction in uraemic patients, direct-acting vasodilators and phosphodiesterase inhibitors will be useful.

Effect of physical training on blood level of endogenous modulators of beta-adreno- and m-cholinoreactivity in patients with a history of myocardial infarction.

The relative content of myocyte-active factors and endogenous muscarinic receptor blocker in the blood increased, while the concentration of endogenous beta-adrenoceptor sensitizer decreased in coronary patients with a history of acute myocardial infarction. Physical training produced a therapeutic effect, normalized the content of these factors and, probably, improved beta-adrenergic and muscarinic cholinergic regulation of the heart and vessels.

Cardiovascular effects of hypocretin-1 in nucleus of the solitary tract.

Experiments were done in male Wistar rats to investigate the effects of microinjection of hypocretin-1 (Hcrt-1) into the nucleus of the solitary tract (NTS) on mean arterial pressure (MAP), heart rate (HR), and the baroreflex. In the first series, the distribution of Hcrt-1-like immunoreactivity (Ir) was mapped within the region of NTS. Hcrt-1 Ir was found throughout the NTS region, predominantly within the caudal dorsolateral (Slt), medial (Sm), and interstitial subnuclei of the NTS. In the second series, in alpha-chloralose or urethane-anesthetized rats, microinjection of Hcrt-1 (0.5-5 pmol) into the caudal NTS elicited a dose-dependent decrease in MAP and HR. A mapping of the caudal NTS region showed that the largest depressor and bradycardia responses elicited by Hcrt-1 were from sites in the Slt and Sm. In addition, doses >2.5 pmol at a small number of sites localized to the caudal commissural nucleus of NTS elicited pressor and tachycardia responses. Intravenous administration of the muscarinic receptor blocker atropine methyl bromide abolished the bradycardia response and attenuated the depressor response, whereas subsequent administration of the nicotinic receptor blocker hexamethonium bromide abolished the remaining MAP response. Finally, microinjection of Hcrt-1 into the NTS significantly potentiated the reflex bradycardia to activation of arterial baroreceptors as a result of increasing MAP by systemic injections of phenylephrine (2-4 microg/kg). These results suggest that Hcrt-1 in the NTS activates neuronal circuits that increases vagal activity to the heart, inhibits sympathetic activity to the heart and vasculature, and alters the excitability of NTS neuronal circuits that reflexly control the circulation.

Acetylcholine receptors do not mediate the immobilization produced by inhaled anesthetics.

Acetylcholine receptors transmit excitatory impulses, are broadly distributed throughout the central nervous system, and are particularly sensitive to the depressant effects of inhaled anesthetics. Thus these receptors are potential mediators of the immobility produced by inhaled anesthetics. We tested this potential in rats by giving intraperitoneal atropine, scopolamine, and mecamylamine to block muscarinic (atropine and scopolamine) and neuronal nicotinic (mecamylamine) acetylcholine receptors. Block with scopolamine (up to 100 mg/kg), atropine (10 mg/kg), mecamylamine (up to 4 mg/kg), or atropine (10 mg/kg) plus mecamylamine (up to 4 mg/kg) did not significantly decrease the isoflurane concentration required to suppress movement to noxious stimulation (minimum alveolar anesthetic concentration). We also gave atropine intrathecally, finding that the infusions that did not cause permanent paralysis produced slight or no decreases in the minimum alveolar anesthetic concentration. We conclude that acetylcholine receptors do not seem to play a role as mediators of immobilization by inhaled anesthetics. Inhaled anesthetics produce two crucial effects: amnesia and immobility in the face of noxious stimulation. Block of muscarinic and neuronal nicotinic acetylcholine receptors in rats does not significantly decrease the isoflurane concentration required to suppress movement to stimulation. Thus, acetylcholine receptors do not seem to play a major role as mediators of the immobilization produced by inhaled anesthetics. Their capacity to mediate other effects of inhaled anesthetics (e.g., amnesia) remains to be tested.

Identification of the biomechanical factors associated with the perception of distension in the human esophagus.

Current techniques used to investigate the mechanisms responsible for the sensory responses to distension of the human esophagus provide limited information because the degree of circumferential stretch required to determine tension can only be inferred. We used impedance planimetry to measure the cross-sectional area during esophageal distension to ascertain the degree of stretch and tension that initiated motor and sensory responses. Hyoscine-N-butyl bromide (HBB), a cholinergic muscarinic receptor blocker, was also used to alter esophageal tension during distension. Motor activity was initiated at a lower degree of stretch and tension than that which initiated sensory awareness; both increased directly with increasing distension. HBB reduced both esophageal motility and tension during distension without altering the relationship between sensation intensity and cross-sectional area. Esophageal stretch, rather than tension, thus appears to be the major factor influencing sensory responses to esophageal distension.

Dextran sodium sulphate-induced colitis perturbs muscarinic cholinergic control of colonic epithelial ion transport.

1. Neuronal cholinergic input is an important regulator of epithelial electrolyte transport and hence water movement in the gut. 2. In this study, colitis was induced by treating mice with 4% (w v(-1)) dextran sodium-sulphate (DSS)-water for 5 days followed by 3 days of normal water. Mid-colonic segments were mounted in Ussing chambers and short-circuit current (Isc, indicates net ion movement) responses to the cholinergic agonist, carbachol (CCh; 10(-4) M)+/-tetrodotoxin, atropine (ATR), hexamethonium (HEX), naloxone or phenoxybenzamine were assessed. 3. Tissues from mice with DSS-induced colitis displayed a drop in Isc in response to CCh (-11.3+/-3.3 microA/cm(2)), while those from control mice showed a transient increase in Isc (76.3+/-13.0 microA/cm(2)). The DeltaIsc in colon from DSS-treated mice was tetrodotoxin-sensitive, atropine-insensitive and was reversed by hexamethonium (HEX+CCh=16.7+/-7.8 microA/cm(2)), indicating involvement of a nicotinic receptor. 4. CCh induced a drop in Isc in tissues from controls only when they were pretreated with the cholinergic muscarinic receptor blocker, atropine: ATR+CCh=-21.3+/-7.0 microA/cm(2). Nicotine elicited a drop in Isc in Ussing-chambered colon from both control and DSS-treated mice that was TTX-sensitive. 5. The drop in Isc evoked by CCh challenge of colonic tissue from DSS-treated mice or ATR+CCh challenge of control tissue was not significantly affected by blockade of opiate or alpha-adrenergic receptors by naloxone or phenoxybenzamine, respectively. 6. The data indicate that DSS-colitis reveals a nicotinic receptor that becomes important in cholinergic regulation of ion transport.

Ectopic discharges trigger sympathetic sprouting in rat dorsal root ganglia following peripheral nerve injury

Experimental backgrounds of ectopic discharges were made by i.p. administrating of 4-aninopyriding (4-AP), a K(+) channel blocker, or anisodamine, a muscarinic receptor blocker, in CCI rats, and the sympathetic sprouting in the dorsal root ganglia (DRG) as well as the heat-hyperalgesia was observed. It was demonstrated that the increased ectopic discharges induced by 4-AP promote sympathetic sprouting in the DRG and a greater number of sympathetic basket cells were developed, causing exacerbation of heat-hyperalgesia in CCI rats. On the contrary, the sympathetic sprouting in the DRG and heat-hyperalgesia are evidently diminished after anisodamine injection. Our results suggest that ectopic discharges may be an immediate factor in triggering sympathetic sprouting in DRG following peripheral nerve injury.

Muscarinic receptors of the neostriatum--their role in controlling operant behavior in dogs.

Chronic experiments were performed on six dogs to study the effects of bilateral microinjections of muscarinic receptor agonists and blockers into the dorsal striatum on the performance of an operant defensive reflex consisting of maintenance of a specified posture and on the differentiation of stimuli. Microinjections of carbachol, a non-selective agonist of muscarinic receptors, were accompanied by increases in the tonic component of movements, inhibition of phasic movements, ordering of the postural rearrangement, and increases in the amplitudes of its components. Bilateral microinjections of the selective agonist oxotremorine into the neostriatum had significantly weaker effects on the amplitude of postural rearrangement, generally decreasing the amplitude. Although oxotremorine also increased the tonic component of the operant response, this effect was weaker than that seen with carbachol microinjections. In addition, oxotremorine, unlike carbachol increased the number of intersignal limb elevations. These data, along with data published in the literature, are used to suggest the hypothesis that the actions of oxotremorine are mediated not only via muscarinic M2 but also via M1 receptors in the neostriatum. Stronger changes in responses to differential stimuli were also obtained after microinjection of the non-selective agonist carbachol than after microinjection of oxotremorine, and the fact that changes in responses to differential stimuli were significantly greater than changes in those to defensive stimuli suggests that microinjections of muscarinic M1 and M4 receptor agonists into the striatum are also accompanied by improvements in attention to significant stimuli.

In Vitro Effects of Organophosphorus Anticholinesterases on Muscarinic Receptor-Mediated Inhibition of Acetylcholine Release in Rat Striatum

The aim of the present study was to evaluate the in vitro modulation of muscarinic autoreceptor function by the organophosphorus (OP) anticholinesterases chlorpyrifos oxon, paraoxon, and methyl paraoxon. Acetylcholine (ACh) release was studied by preloading slices from rat striatum with [3H]choline and depolarizing with potassium (20 mM) in perfusion buffer containing hemicholinium-3 (to prevent reuptake of radiolabeled choline). Under these conditions, chlorpyrifos oxon, paraoxon, and methyl paraoxon (0.1–10 μM) all reduced ACh release in a concentration-dependent manner. Addition of the carbamate acetylcholinesterase (AChE) inhibitor physostigmine (20 μM) to the perfusion buffer also decreased ACh release. When physostigmine was present, the three oxons had no additional effect on ACh release. Concentration-dependent inhibition of AChE activity in striatal slices perfused with chlorpyrifos oxon (0.1, 1, and 10 μM) suggested AChE inhibition was responsible for oxon-mediated alterations in ACh release. To differentiate between direct and indirect actions of the OP toxicants on muscarinic autoreceptors, we compared the effects of the oxons on ACh release under two conditions, i.e., tissues were perfused with buffer containing only hemicholinium-3 or with buffer containing hemicholinium-3, physostigmine, and the nonselective muscarinic receptor blocker atropine (100 nM). In the presence of only hemicholinium-3, concentration-dependent inhibition of ACh release was again noted for all oxons, similar to the effects of the muscarinic agonists carbachol and cis-dioxolane. In the presence of physostigmine and atropine, the relative potencies of all agents were markedly reduced. Interestingly, carbachol, cis-dioxolane, paraoxon, and methyl paraoxon all decreased ACh release as before, but chlorpyrifos oxon (100–300 μM) actually increased ACh release. Together, the results suggest that chlorpyrifos oxon, paraoxon, and methyl paraoxon can activate muscarinic autoreceptors indirectly through inhibition of AChE. Both paraoxon and methyl paraoxon also directly activate whereas chlorpyrifos oxon blocks muscarinic autoreceptor function. Qualitative differences in the direct actions of these oxons at this presynaptic regulatory site could contribute to differential toxicity with high-dose exposures.

The Effects of L-glucose on memory in mice are modulated by peripherally acting cholinergic drugs.

D-Glucose improves memory in animals and humans and in subjects with memory pathologies. To date, the accepted conclusion drawn from animal research is that D-glucose improves memory via alterations in central cholinergic systems. However, recent evidence suggests that a sugar which does not cross the blood-brain barrier also facilitates memory (Talley, Arankowsky-Sandoval, McCarty, & Gold, 1999). The present study examined the effects of peripherally administered L-glucose, a stereoisomer of D-glucose, in male mice. Intraperitoneal administration of L-glucose (300 mg/kg) before testing enhanced place learning in the Morris water maze. Mice injected with L-glucose had significantly shorter escape latencies than mice injected with saline (1 ml/kg). Effects were observed on both reference memory and working memory tasks. L-Glucose did not facilitate performance on either task when it was simultaneously administered with cholinergic antagonists that are excluded from the central nervous system. Thus, simultaneous administration of either methyl-scopolamine (0.3 mg/kg), a peripherally acting muscarinic receptor blocker, or hexamethonium (1 mg/kg), a peripherally acting nicotinic receptor blocker, reversed the effect of L-glucose on memory. These findings suggest that the memory effects of l-glucose may be mediated by facilitated acetylcholine synthesis and/or release in the peripheral nervous system.

Functional relationship between subfornical organ cholinergic stimulation and cellular activation in the hypothalamus and AV3V region

The subfornical organ (SFO) has been suggested to be important for water intake and secretion of vasopressin (AVP). However, the role of the SFO cholinergic mechanism in the control of body fluid regulation is not clear. This study determined the effects of local cholinergic stimulation in the SFO produced by administration of physostigmine on drinking and cellular excitation in the anterior third ventricle (AV3V) region and in the supraoptic and paraventricular nuclei (SON and PVN). The results showed that injection of physostigmine into the SFO induced water intake and c-fos expression in the AV3V area as well as in the AVP containing neurons in the hypothalamus. Pretreatment of the SFO with mecamylamine, a nicotinic receptor antagonist, had no effect on physostigmine induced behavioral and c-fos responses. The muscarinic receptor blocker atropine, however, abolished both drinking and cellular activation after injection of physostigmine into the SFO. Immunostaining experiments demonstrated positive acetyltransferase (ChAT) in the SFO. Intensive ChAT immunoreactivity was located in the cholinergic fibers in the SFO. Together, the results indicate that SFO cholinergic mechanisms are important in co-operation with the AV3V and hypothalamic neurons in the control of thirst and AVP-mediated body fluid homeostasis.

Memory improving actions of gabapentin in mice: possible involvement of central muscarinic cholinergic mechanism

Male CF-1 mice were tested 48 h after training on a one trial step-through inhibitory avoidance task. Immediately post-training, intraperitoneal (i.p.) injections of the antiepileptic gabapentin (1-(aminomethyl) cyclohexaneacetic acid) (GBP, 10 mg/kg) enhanced retention performance. The effect was prevented by atropine, a central muscarinic cholinergic receptor antagonist (0.5 mg/kg, i.p.) administered after training but 10 min prior to GBP treatment. In contrast, neither methylatropine (0.5 mg/kg, i.p.), a peripherally acting muscarinic receptor blocker, nor mecamylamine (5 mg/kg, i.p.) or hexamethonium (5 mg/kg, i.p.), two cholinergic nicotinic receptor antagonists, prevented the effects of post-training GBP on retention performance. Low subeffective doses of the central acting anticholinesterase physostigmine (35 mg/kg, i.p.) administered immediately after training, and GBP (5 mg/kg, i.p.), given 10 min after training, significantly enhanced retention performance. The effects of GBP (5 mg/kg, i.p.) were not influenced by the peripherally acting anticholinesterase neostigmine (150 mg/kg, i.p.). Considered together, these findings suggest a disinhibitory action of GBP on the activity of central muscarinic cholinergic mechanisms that are involved in memory consolidation.

Rapid differential conditioning of the somatosensory evoked potential by changed patterns of brief innocuous tactile stimuli in waking rats is altered by atropine sulfate

Air puffs delivered to the nose of an awake, lightly restrained rat every 15 s produced evoked potentials that changed gradually over time so that the averaged response to the last 40 stimuli was measurably different from the first 40. This habituation-like paradigm increased the size of an early component of the potential in several places. When measured with respect to the time of stimulus onset (there was a 21.6 ms delay in the time of arrival of the stimulus maximum at the nose), one of the largest increases occurred 46 ms later (39 ms latency to onset, and 55 ms latency to offset). As well, a late component of the waveform became more positive, showing a maximum between 156 and 185 ms (133 ms latency to onset, and more than 250 ms latency to offset). Changing the pattern but not the number of stimuli accelerated the rate of this positive shift with a maximum at 37 ms (21 ms latency to onset, and 42 ms latency to offset), but did not affect the rate of change in the late component. This effect of altering the temporal pattern of the stimuli was blocked by systemic injections of atropine sulfate, a blocker of central muscarinic receptors, whereas, neither saline injections nor atropine methyl nitrate injections (an atropine analog that does not cross the blood–brain barrier) could produce these changes. These observations suggest that the adaptive changes of the somatosensory evoked potential induced by novel patterns intercalated in otherwise monotonous repetitive somatic stimuli depend upon central muscarinic mechanisms.

The role of cholinergic and peptidergic pathways in the regulation of pancreatic exocrine function during postnatal development in pigs.

The aim of this study was to investigate the role of the parasympathetic (cholinergic and peptidergic) nervous system in the regulation of exocrine pancreas function in piglets during their early postnatal development. The cholinergic and peptidergic regulatory pathways of exocrine pancreatic function were tested by the specific muscarinic receptor blocker 4-diphenylacetoxy-N-methylpiperidine-methiodide (4-DAMP) and bombesin, respectively. At the age of 2 weeks, piglets were surgically fitted with a chronic pancreatic duct catheter, a duodenal re-entrant cannula and a jugular vein catheter. The experiments comprised a pre-weaning period, and a post-weaning period that commenced at the beginning of the 5th week of age. Intravenous infusion of 4-DAMP (100 pmol x kg(-1) x h(-1)) reduced the outflow of pancreatic juice, the output of total protein and the activity of trypsin, chymotrypsin, carboxyl ester hydrolase and amylase during preprandial and postprandial pancreatic secretion, in both the pre- and post-weaning periods. However, the inhibitory effect of 4-DAMP during postprandial secretion was significantly greater (P < 0.05) in suckling piglets. The infusion of bombesin (10, 100 and 1000 pmol x kg(-1) x h(-1)) stimulated exocrine pancreatic secretion in a dose-dependent manner during both the pre- and post-weaning periods. However, the stimulatory effect of 1000 pmol x kg(-1) x h(-1) bombesin on total protein output and the activities of trypsin, chymotrypsin and amylase were significantly higher (P < 0.05) in suckling piglets. In summary, our study showed that cholinergic and peptidergic mechanisms are involved in the regulation of exocrine pancreas function in piglets in both the pre- and post-weaning stages. 4-DAMP had a greater inhibitory effect on exocrine pancreatic secretion in piglets during the pre-weaning period. Thus, these observations suggest that the parasympathetic nervous system plays a dominant role in the functioning of the exocrine pancreas at this time. The action of bombesin suggests that it is a potent secretagogue for the exocrine pancreas in pigs during their postnatal development.

Adrenergic and cholinergic mechanisms of hemopoiesis regulation during experimental neuroses.

Scopolamine abolished hyperplasia of erythropoiesis caused by conflict situation and its inhibition after paradoxical sleep deprivation, but did not affect granulomonocytopoiesis. The modulatory effects of this muscarinic receptor blocker on the erythron indicated involvement of the cholinergic system in the formation of neurotransmitter interrelations. In vitro restoration of the colony-forming ability of myelokaryocytes under the effect of sympathomimetics during in vivo blockade of the central or peripheral part of the adrenergic system indicated that adrenoceptors play the major role in the information transfer from the higher regulatory centers to hemopoietic cells.

The Amygdala Is Involved in the Modulation of Long-Term Memory, but Not in Working or Short-Term Memory

Rats with cannulae implanted in the junction between the central and the basolateral nuclei of the amygdala were trained in one-trial step-down inhibitory avoidance and tested at 3 s for working memory (WM) or 1.5 or 24 h later for short-term memory (STM) and long-term memory (LTM), respectively. Several drugs were infused 6 min prior to training in the animals in which WM was measured or 0 min posttraining in those in which STM and LTM were measured: the glutamate receptor antagonists CNQX (0.5 μg) and AP5 (5.0 μg), the indirect GABAAreceptor antagonist picrotoxin (0.08 μg), the cholinergic muscarinic receptor blocker scopolamine (2.0 μg), norepinephrine (0.3 μg), the protein kinase C inhibitor staurosporin (1.0 μg), or the calcium/calmodulin dependent protein kinase II inhibitor Kn-62 (3.5 ng). None of the drugs had any effect on either WM or STM. All had, as previously shown, strong effects on LTM: picrotoxin and norepinephrine enhanced it, and CNQX, AP5, scopolamine, Kn-62, and staurosporin inhibited it. The results do not support the idea that memory of this task is formed in the amygdala; they indicate that the amygdala is not involved in WM or STM processing and support the idea that the amygdala modulates LTM storage processes carried out elsewhere.

Enhanced sensitivity of the iris sphincter to the muscarinic agonist carbachol at lower temperature.

Cooling of the human iris sphincter from 37.5 degrees C to 17.5 degrees C potentiates the concentration response curves of the muscarinic agonist, carbachol. Although EC50 value 0.31 micromol/l of the agonist was not significantly affected at the lower temperature, the maximum contraction of the control at 37.5 C was potentiated by 263%. Such potentiation was 272%, 135% and 125% for the dog, horse and pig in sphincter, respectively. Low temperature potentiation of the carbachol can be reversed by warming the tissue to 37.5 degrees C. Pretreatment of the tissue with competitive muscarinic receptor blocker, atropine, blocked the contractile action of carbachol at 37.5 degrees C and 17.5 degrees C which resulted in nearly equal KB of approximately 2 nmol/l, but at 17.5 degrees C, the contraction induced by carbachol was antagonized by atropine with difficulty to give pseudo KB of 67 nmol/l. Ideally, a competitive reversible antagonist is expected to give equal KB values for the drug receptor interaction; irrespective of the method of study. The higher KB value indicates a functional antagonism of the agonist activated cascade by the blocker. Clinical implications of the drug antagonism at low temperatures are discussed.

Effects of Posttraining Administration of Insulin on Retention of a Habituation Response in Mice: Participation of a Central Cholinergic Mechanism

Male Swiss mice were allowed to explore a novel environment, provided by an open-field activity chamber for a 10-min period. The procedure was repeated twice within a 24-h interval. The difference in the exploratory activity between the first (training) and the second exposure (testing) to the chamber was taken as an index of retention of this habituation task. Posttraining intraperitoneal administration of insulin (8, 20, or 80 IU/kg) impaired retention in a dose-related manner, although only the dose of 20 IU/kg of insulin produced significant effects. Thus, the dose–response curve adopted a U-shaped form. Insulin (20 IU/kg) given to untrained mice did not modify their exploratory performance when recorded 24 h later. The effects of insulin on retention were time dependent, suggesting an action on memory storage. An ineffective dose (8 IU/kg) of insulin given together with an ineffective dose of a central acting muscarinic cholinergic antagonist atropine (0.5 mg/kg) or with a central acting nicotinic cholinergic antagonist mecamylamine (5 mg/kg) interacted to impair retention. In contrast, neither methylatropine (0.5 mg/kg), a peripherally acting muscarinic receptor blocker, nor hexamethonium (5 mg/kg), a peripherally acting nicotinic receptor blocker, interacted with the subeffective dose of insulin on retention. The impairing effects of insulin (20 IU/kg) on retention were reversed by the simultaneous administration of physostigmine (70 μg/kg) but not neostigmine (70 μg/kg). We suggest that insulin impairs memory storage of one form of learning elicited by stimuli repeatedly presented without reinforcement, probably through a decrement of brain acetylcholine synthesis.