Biomedical subjects

H Fukuta

Publications and source records attributed to H Fukuta.

At least 19 recordsLinked to original sources

PubMed2003

Mechanisms of excitatory transmission in circular smooth muscles of the guinea pig seminal vesicle.

PURPOSE: Cellular mechanisms of excitatory neuromuscular transmission in circular smooth muscles of the seminal vesicle were investigated. MATERIALS AND METHODS: Circular smooth muscles of the seminal vesicle of the guinea pig were isolated. Changes in membrane potential produced by transmural nerve stimulation were recorded using intracellular microelectrode techniques. Changes in the intracellular Ca ion concentration induced by transmural nerve stimulation were measured in preparations loaded with Ca indicator fura-PE3. Responses produced by bath applied norepinephrine and alpha,beta-methylene adenosine triphosphate (ATP) were also examined. RESULTS: Transmural nerve stimulation evoked excitatory junction potentials that triggered action potentials and also caused transient increases in [Ca2+] (Ca transients). Nifedipine abolished action potentials, leaving underlying excitatory junction potentials unchanged, and reduced the amplitude of Ca transients. Excitatory junction potentials were blocked by alpha,beta-methylene ATP or guanethidine but not by phentolamine. A train of transmural nerve stimulation evoked oscillatory changes in membrane potential and [Ca2+], which were abolished by phentolamine or inhibited by nifedipine. Nifedipine insensitive components were abolished by cyclopiazonic acid. Norepinephrine depolarized the membrane and elicited oscillatory potentials with an associated elevation in [Ca2+]. These responses were inhibited by nifedipine and abolished by additional application of cyclopiazonic acid. Transient depolarization with an associated increase in [Ca2+] was elicited by alpha,beta-methylene ATP and [Ca2+] responses but no potential changes were inhibited by nifedipine. CONCLUSIONS: Circular smooth muscles of the guinea pig seminal vesicle receive a projection of sympathetic nerves that release norepinephrine to initiate slow depolarization through the activation of alpha-adrenoceptors. These nerves also release ATP to elicit excitatory junction potentials. Neurally released norepinephrine and ATP are increased [Ca2+] by the influx of Ca2+ through L-type Ca2+ channels and also by the release of Ca2+ from internal stores.

Action Potentials↗

PubMed2002

Modulators of internal Ca2+ stores and the spontaneous electrical and contractile activity of the guinea-pig renal pelvis.

1. The role of internal Ca(2+) stores in the generation of the rhythmic electrical and contractile activity in the guinea-pig proximal renal pelvis was examined using intracellular microelectrode and muscle tension recording techniques. 2. Ryanodine (30 microM) transiently increased contraction amplitude, while caffeine (0.5 - 3 mM) reduced contraction amplitude and frequency. Contractility was also reduced by 2-aminoethoxy-diphenylborate (2-APB 60 microM), xestospongin C (1 microM), U73122 (5 microM) and neomycin (4 mM), blockers of IP(3)-dependent release from Ca(2+) stores. 3. 60 mM K(+) saline-evoked contractions were reduced by caffeine (1 mM), U73122 (5 microM) and neomycin (4 mM), but little affected by ryanodine or 2-APB (60 microM). 4. Spontaneous action potentials consisting of an initial spike followed by a long plateau were recorded (frequency 8.6+/-1.0 min(-1)) in small urothelium-denuded strips of proximal renal pelvis. 5. Action potential discharge was blocked in 75 and 35% of cells by 2-APB (60 microM) and caffeine (1 mM), respectively. In the remaining cells, only a truncation of the plateau phase was observed. 6. Cyclopiazonic acid (CPA 10 microM for 10 - 180 min), blocker of CaATPase, transiently increased contraction frequency and amplitude. Action potential durations were increased 3.6 fold. Contraction amplitude and frequency slowly declined during a prolonged (>60 min) CPA exposure. 7. We conclude that the action potential in caffeine-sensitive cells and the shoulder component of caffeine-insensitive action potential arise from the entry of Ca(2+) through Ca(2+) channels. The inhibitory actions of modulators of internal Ca(2+) release were partially explained by a blockade of Ca(2+) entry.

Animals↗

PubMed2001

Origin and propagation of spontaneous excitation in smooth muscle of the guinea-pig urinary bladder.

The origin and propagation of waves of spontaneous excitation in bundles of smooth muscle of the guinea-pig bladder were examined using intracellular recording techniques and visualization of the changes in the intracellular calcium concentration ([Ca2+]i). Bladder smooth muscle cells exhibited spontaneous transient increases in [Ca2+]i which originated along a boundary of each smooth muscle bundle and then spread to the other boundary with a conduction velocity of 2.0 1r1r>mm1> s-1. Spontaneous increases in [Ca2+]i were always preceded by action potentials. Nifedipine (10 microM) abolished increases in both [Ca2+]i and action potentials. Caffeine (10 1s1sFmM1F), ryanodine (50 microM) and cyclopiazonic acid (10 microM reduced the amplitude of the associated increases in [Ca2+]i without preventing the generation of action potentials. Spontaneous action potentials had conduction velocities of 40 1t1t>mm 1> s-1 in the axial direction and 1.3 1u1u>mm 1> s-1 in the transverse direction. The electrical length constants of the bundles of muscle were 425 microM in the axial direction and 12.5 microM in the transverse direction. Neurobiotin, injected into an impaled smooth muscle cell, spread more readily to neighbouring cells located in the axial direction than those located in the transverse direction. The spread of neurobiotin was inhibited by 18beta-glycyrrhetinic acid (18beta-GA, 40 microM), a gap junction blocker. Immunohistochemistry for Connexin 43 showed abundant punctate staining on the smooth muscle cell membranes. These results suggested that spontaneous action potentials and associated calcium waves occur almost simultaneously along the boundary of bladder smooth muscle bundles and then propagate to the other boundary probably through gap junctions.

Action Potentials↗

PubMed2001

Postural response of low-frequency component of heart rate variability is an increased risk for mortality in patients with coronary artery disease.

STUDY OBJECTIVES: We examined whether autonomic functions assessed by heart rate variability (HRV) during standardized head-up tilt testing (HUTT) predict risk for death in stable patients with coronary artery disease (CAD). DESIGN AND SETTING: Retrospective cohort study in medium-sized university general hospital. MEASUREMENTS AND RESULTS: In a cohort of 250 patients with CAD who were undergoing elective coronary angiography, we analyzed HRV during standardized HUTT under paced breathing with discontinuation of treatment with all medications. During a subsequent mean follow-up period of 99 months, there were 13 cardiac deaths and 12 noncardiac deaths. Cox regression analysis adjusted for cardiovascular risks revealed that increased postural change (supine to upright) in the power of low-frequency component (LF) power predicted an increased risk for cardiac death (relative risk [per 1-ln ms(2) increment], 4.36; 95% confidence interval, 1.64 to 11.6), while neither the high-frequency component nor its response to HUTT predicted any form of death. When the patients were trichotomized by the level of postural LF change (large drop, < or = - 0.6 ln[ms(2)]; small drop and rise, > 0 ln[ms(2)]), the three groups did not differ in terms of clinical features or CAD severity at baseline or coronary interventions during the follow-up period; however, the 8-year cardiac mortality rates were 0%, 6%, and 12%, respectively (p = 0.008 [log rank test]). Additionally, the difference was enhanced when analyzed excluding 64 patients who had been treated with a beta-blocker during the follow-up period (0%, 7%, and 15%, respectively; p = 0.006 [log rank test]). CONCLUSIONS: The postural response of HRV predicts the risk for death in patients with CAD. Postural LF increase (LF rise), in particular, is an independent risk factor for cardiac death.

Adult↗

PubMed2001

Effects of isoproterenol on spontaneous excitations in detrusor smooth muscle cells of the guinea pig.

PURPOSE: Because beta-adrenoceptor agonists would be a useful tool for the pharmacological treatment of unstable bladder, we investigated the cellular mechanisms underlying beta-adrenoceptor mediated inhibition on spontaneous excitation in detrusor smooth muscle. MATERIALS AND METHODS: Detrusor smooth muscle bundles were isolated from guinea pig bladders. Changes in membrane potential were recorded using an intracellular recording technique. In preparations loaded with the calcium indicator fura-PE3 changes in the concentration of intracellular calcium ions were measured simultaneously with membrane potential. Effects of isoproterenol on spontaneous changes in the membrane potential and intracellular Ca(2+) were examined RESULTS: Detrusor smooth muscle cells exhibited spontaneous action potentials that were associated with transient increases in intracellular Ca(2+) (calcium transients). Isoproterenol, which hyperpolarized the membrane, prevented action potentials and calcium transients. This induced inhibition of calcium transients was not affected by cyclopiazonic acid. Isoproterenol induced hyperpolarization was inhibited by inhibitors of protein kinase A, N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide, hydrochloride and Rp-adenosine-3',5'-cyclic phosphorothioate. Hyperpolarization was blocked by a solution containing 30 mM. potassium but not by a range of potassium channel blockers. Ouabain and a solution of 0.5 mM. potassium also inhibited hyperpolarization. CONCLUSIONS: Our results suggest that isoproterenol prevented spontaneous action potential discharges and associated calcium transients through the activation of protein kinase A. The isoproterenol induced inhibition of intracellular Ca(2+) largely depends on the prevention of spontaneous action potentials since the contribution of the intracellular calcium store was small. Isoproterenol hyperpolarizes the membrane, probably by stimulating sodium pump activity.

Action Potentials↗

PubMed2000

Arginine kinase from Nautilus pompilius, a living fossil. Site-directed mutagenesis studies on the role of amino acid residues in the Guanidino specificity region.

Arginine kinases were isolated from the cephalopods Nautilus pompilius, Octopus vulgaris, and Sepioteuthis lessoniana, and the cDNA-derived amino acid sequences have been determined. Although the origin and evolution of cephalopods have long been obscure, this work provides the first molecular evidence for the phylogenetic position of Cephalopoda in molluscan evolution. A crystal structure for Limulus arginine kinase showed that four amino acid residues (Ser(63), Gly(64), Val(65), and Tyr(68)) are hydrogen-bonded with the substrate arginine. We introduced three independent mutations, Ser(63) --> Gly, Ser(63) --> Thr, and Tyr(68) --> Ser, in Nautilus arginine kinase. One of the mutants had a considerably reduced substrate affinity, accompanied by a decreased V(max). In other mutants, the activity was lost almost completely. It is known that substantial conformational changes take place upon substrate binding in arginine kinase. We hypothesize that the hydrogen bond between Asp(62) and Arg(193) stabilizes the closed, substrate-bound state. Site-directed mutagenesis studies strongly support this hypothesis. The mutant (Asp(62) --> Gly or Arg(193) --> Gly), which destabilizes the maintenance of the closed state and/or perhaps disrupts the unique topology of the catalytic pocket, showed only a very weak activity (0.6-1.5% to the wild-type).

Amino Acid Sequence↗

PubMed2000

Passive terminal extension causes anterior tibial translation in some anterior cruciate ligament-deficient knees.

The purpose of the present study was to accurately measure anterior tibial translation during passive terminal extension (ATT-PTE) in anterior cruciate ligament (ACL)-deficient knees, and to investigate correlations between various characteristics of such knees and the magnitude of ATT-PTE. The subjects were 79 patients with unilateral ACL-deficient knees and little flexion contracture. All patients were confirmed to have ACL injury of one knee by arthroscopy. Lateral radiographs of the bilateral knees in passive terminal extension were compared, and ATT-PTE was measured using an original superimposition method. The inter-observer and intra-observer reproducibility of measurement was significantly greater for this method than for the method without superimposition. In 42 of the 79 patients (53%), ATT-PTE was greater than 1 mm, while it was greater than 4 mm in 15 patients (19%). ATT-PTE was significantly larger in patients with a large anterior displacement difference (as measured with the KT-1000 arthrometer) (ATT-KT), a long period since injury, a history of reinjury, injury to the meniscus, and the presence of gross pivot shift. On the other hand, ATT-KT was equal to or greater than 2.5 mm in all 79 patients and showed no significant correlation with the time since initial injury, history of reinjury, on injury to the meniscus.

Adolescent↗

PubMed2000

Hyperpolarization-induced dilatation of submucosal arterioles in the guinea-pig ileum.

1. The effects of inhibition of acetylcholine (ACh)-induced hyperpolarization on dilatation of submucosal arterioles were investigated in the guinea-pig ileum. 2. In smooth muscles of the arterioles depolarized by Ba(2+) (0.5 mM) to about -40 mV, ACh (3 microM) repolarized the membrane to about -65 mV (hyperpolarization), irrespective of the absence or presence of L-N(omega)-nitroarginine (L-NOARG, 0.1 mM) and diclofenac (1 microM), and increased the diameter (dilatation). 3. Combined application of charybdotoxin (CTX, 50 nM) and apamin (0.1 microM), inhibitors of some types of K(+)-channels, abolished the ACh-induced hyperpolarization and dilatation. 4. 18 beta-Glycerrhetinic acid (18 beta-GA, 30 microM), a known inhibitor of gap junctions, depolarized the membrane to about -36 mV, either in the absence or in the presence of Ba(2+), with no associated contraction of the arterioles. In the presence of 18 beta-GA, ACh-induced hyperpolarization was abolished, however the dilatation was inhibited only partially, with associated inhibition of constriction produced by Ba(2+) and NA. 5. 18 beta-GA inhibited the dilatation produced by sodium nitroprusside, an NO donor. 6. The ACh-induced hyperpolarization and dilatation were abolished in the presence of 2-aminoethoxydiphenyl borate (30 microM), an inhibitory modulator of inositol trisphosphate receptor-mediated Ca(2+) release from intracellular stores. 7. It is concluded that in submucosal arterioles, hyperpolarizations produced by ACh have causal relationship to the arteriolar dilatation. 18 beta-GA did not induce parallel relationship between hyperpolarization and dilatation produced by ACh. 18 beta-GA may have unidentified inhibitory effects on agonist-mediated actions, in addition to the inhibition of gap junctions.

Acetylcholine↗

PubMed1999

Calcium responses induced by acetylcholine in submucosal arterioles of the guinea-pig small intestine.

1. Calcium responses induced by brief stimulation with acetylcholine (ACh) were assessed from the fluorescence changes in fura-2 loaded submucosal arterioles of the guinea-pig small intestine. 2. Initially, 1-1.5 h after loading with fura-2 (fresh tissues), ACh increased [Ca2+]i in a concentration-dependent manner. This response diminished with time, and finally disappeared in 2-3 h (old tissues). 3. Ba2+ elevated [Ca2+]i to a similar extent in both fresh and old tissues. ACh further increased the Ba2+-elevated [Ca2+]i in fresh tissues, but reduced it in old tissues. Responses were not affected by either indomethacin or nitroarginine. 4. In fresh mesenteric arteries, mechanical removal of endothelial cells abolished the ACh-induced increase in [Ca2+]i, with no alteration of [Ca2+]i at rest and during elevation with Ba2+. 5. In the presence of indomethacin and nitroarginine, high-K+ solution elevated [Ca2+]i in both fresh and old tissues. Subsequent addition of ACh further increased [Ca2+]i in fresh tissues without changing it in old tissues. 6. Proadifen, an inhibitor of the enzyme cytochrome P450 mono-oxygenase, inhibited the ACh-induced changes in [Ca2+]i in both fresh and Ba2+-stimulated old tissues. It also inhibited the ACh-induced hyperpolarization. 7. In fresh tissues, the ACh-induced Ca2+ response was not changed by apamin, charybdotoxin (CTX), 4-aminopyridine (4-AP) or glibenclamide. In old tissues in which [Ca2+]i had previously been elevated with Ba2+, the ACh-induced Ca2+ response was inhibited by CTX but not by apamin, 4-AP or glibenclamide. 8. It is concluded that in submucosal arterioles, ACh elevates endothelial [Ca2+]i and reduces muscular [Ca2+]i, probably through the hyperpolarization of endothelial or smooth muscle membrane by activating CTX-sensitive K+ channels.

4-Aminopyridine↗

PubMed1999

Inhibition of the endothelium-dependent relaxation by 18beta-glycyrrhetinic acid in the guinea-pig aorta.

The effect of 18beta-glycyrrhetinic acid (GA), an agent which interferes with gap junction conductivity, on endothelium-dependent relaxation produced by substance P was investigated in isolated aortic rings of the guinea-pig. In nor-adrenaline (NA)-contracted aortic rings, substance P (10(-7) M) induced an endothelium-dependent, transient relaxation. The relaxation was only slightly reduced by the co-application of nitroarginine and diclofenac. When GA (2x10(-5) M) was applied first, it slightly reduced substance P-induced relaxation, and a subsequent co-application of nitroarginine and diclofenac strongly reduced the relaxation. In aortic rings contracted with high-K solution ([K(+)](o) = 29.4 mM), substance P-induced relaxation was reduced by the simultaneous application of GA, nitroarginine and diclofenac, but not by GA alone. In endothelium-denuded aortic rings, GA reduced the threshold concentration of NA required to produce contractions and increased the amplitude of NA-induced contractions. GA increased the amplitude of contraction produced by small increases of [K(+)](o) (<30 mM) but reduced those produced by higher concentrations of [K(+)](o) (>54 mM). In NA-contracted aortic rings, Y-26763, a K(+)-channel opener, could relax muscles with reduced amplitude in the presence of GA. It is concluded that in guinea-pig aortic rings, GA inhibits mainly the EDHF-induced components of endothelium-dependent relaxation. GA also modulated contractions produced by NA or high-K solutions. The possible effects of inhibition of gap junctions by GA on endothelium-dependent relaxation were discussed.

Administration, Topical↗

PubMed1998

Blockade by 18beta-glycyrrhetinic acid of intercellular electrical coupling in guinea-pig arterioles.

1. Intercellular electrical communication between smooth muscle and endothelial cells was examined in guinea-pig mesenteric arterioles using the whole-cell patch-clamp method. The time course of the current required to impose a 10 mV voltage clamp step was used to determine the extent of electrical coupling between them. Currents recorded from both smooth muscle and endothelial cells relaxed in a multi-exponential manner, indicating the existence of electrical coupling between cells. 2. 18beta-Glycyrrhetinic acid, a gap junction blocker, quickly blocked electrical communication at 40 microM, while neither heptanol nor octanol did so at concentrations of up to 1 mM. 3. In the current clamp mode, repetitive spikes, induced by 10 mM Ba2+ solutions, could be recorded from both kinds of cells. After blocking gap junctions, spikes could only be recorded from the smooth muscle cell layer, indicating that they had been conducted through myoendothelial junctions. 4. In endothelial cells, acetylcholine (ACh, 3 microM) induced hyperpolarizing responses, which had two phases (an initial fast and a second slower phase) in the current clamp condition. This ACh response persisted in the presence of 18beta-glycyrrhetinic acid, although this compound seemed to make the membrane slightly leaky. 5. After blocking gap junctions, the membrane potential of a single cell in a multicellular preparation could be well clamped. Thus, 18beta-glycyrrhetinic acid may be useful in studying the function of both arteriolar smooth muscle and endothelial cells while they remain located within a multicellular preparation.

Acetylcholine↗

PubMed1998

Potassium channels activated in the endothelium-dependent hyperpolarization in guinea-pig coronary artery.

1. Properties of endothelium-dependent hyperpolarization evoked by acetylcholine (ACh) in smooth muscle of the guinea-pig coronary artery were investigated using conventional microelectrode techniques. 2. ACh hyperpolarized the membrane in an endothelium-dependent manner. The hyperpolarization comprised two components: an initial and a slow hyperpolarization. The former appeared during application of ACh, while the latter occurred after withdrawal of ACh. 3. Indomethacin and f1p4ofenac, inhibitors of the enzyme cyclo-oxygenase, blocked only the slow hyperpolarization, indicating that this potential was produced by endothelial prostanoids. 4. Clotrimazole and SKF 525a, known inhibitors of the enzyme cytochrome P450, inhibited both the initial and the slow hyperpolarizations, suggesting that these chemicals acted as non-selective inhibitors of arachidonic acid metabolism. Inhibition of the lipoxygenase pathway of arachidonic acid metabolism by nordihydroguaiaretic acid had no effect on either component of the hyperpolarization. 5. The slow hyperpolarization was inhibited by 4-aminopyridine (4-AP; 10(4) 10(-3) M) and glibenclamide (10(-6) M). The initial hyperpolarization was greatly inhibited by charybdotoxin (CTX; 5 x 10(-8) M) and partially inhibited by apamin (10(-7) M), but was not inhibited by glibenclamide (10(-5) M). Ba2+ (10(-4) M) depolarized the membrane and increased the amplitude of both components of the ACh-induced hyperpolarization. 6. Hyperpolarizations produced by Y-26763, a K+ channel opener, were inhibited by glibenclamide, but not by 4-AP. 7. The results indicate that the slow hyperpolarization is produced by endothelial prostanoids through activation of 4-AP-sensitive K+ channels (possibly delayed rectifier type). The initial hyperpolarization is produced mainly through activation of CTX-sensitive K+ channels (possibly Ca(2+)-sensitive type).

Acetylcholine↗

PubMed1998

[Endothelium-derived hyperpolarizing factor and vasodilatation].

Properties of endothelium-derived hyperpolarizing factor (EDHF) have been reviewed briefly. The production of EDHF requires an increase in endothelial [Ca2+]i, the properties being similar to those of nitric oxide (NO). EDHF activates K(+)-channels and hyperpolarizes vascular smooth muscle. The EDHF-induced hyperpolarization is greatly inhibited by charybdotoxin (ChTX) and partially inhibited by apamin, but not by K(+)-channel inhibitors such as Ba2+, glibenclamide, 4-aminopyridine, suggesting that the K(+)-channels involved are mainly the Ca(2+)-sensitive type. Membrane hyperpolarization induces vasodilation by unidentified mechanisms. Experiments using K(+)-channel openers and electrophysiology suggest that hyperpolarization may reduce (i) influx of Ca2+ through voltage-sensitive Ca(2+)-channels, (ii) production of InsP3 in the case of agonist-induced contraction, (iii) Ca(2+)-sensitivity of contractile elements and (iv) agonist-stimulated ion channel activities. In the endothelium-dependent vasodilation, the EDHF/EDRF ratio is larger in peripheral vessels than in the proximal ones, indicating significant importance of EDHF mainly in peripheral arteries. The chemical nature of EDHF remains undetermined, although some candidates such as arachidonic acid metabolites or endogenous cannabinoids are proposed. As the inhibition of gap junctions in artrerial tissues reduces the amplitude of EDHF-induced relaxation, the possible involvement of electrical communication between endothelial and smooth muscle cells has also been considered.

Animals↗

PubMed1997

Inhibitory actions of endothelial products on mechanical and calcium responses in aortic smooth muscle cells.

The effects of endothelial products on intracellular calcium ion concentrations ([Ca2+]i) were investigated in the guinea-pig aorta. The perfusate of bradykinin-stimulated cultured endothelial cells relaxed aortic rings by approximately 70%; it was reduced to approximately 50% by nitroarginine, to approximately 30% in high-K solution and remained unaltered by indomethacin. The perfusate elevated the cyclic GMP content in the muscle, which was inhibited by nitroarginine. In cultured, aortic muscle cells, bradykinin elevated [Ca2+]i with an initial transient and following sustained phase; the former was absent after treatment with cyclopiazonic acid while the latter was abolished in [Ca2+]o-free medium. The perfusate lowered aortic [Ca2+]i, and this action was weakened by nitroarginine and diminished in high-K solution. Therefore, the perfusate reduced aortic [Ca2+]i with and without an increase in cyclic GMP production. These actions were sensitive to nitroarginine and high-K, respectively, suggesting that the perfusate contained at least two relaxants, EDRF and EDHF, with both lowering [Ca2+]i in aortic muscle mainly by inhibiting Ca2+ influx.

Animals↗

PubMed1997

Comparison of the relaxing actions of acetylcholine and substance P in smooth muscle of the guinea-pig aorta.

The relationship between relaxation produced by acetylcholine (ACh) or substance P (SP) and tissue cyclic GMP content was investigated in the isolated guinea-pig aorta. ACh and SP relaxed aortic rings precontracted with noradrenaline (NA) or high-K solution ([K+]o = 38.8 mM), in an endothelium-dependent manner. The amplitude of relaxation was larger for SP than for ACh. Nitroarginine inhibited ACh-induced but not SP-induced relaxation in NA-contraction, while this chemical inhibited both ACh- and SP-induced relaxations in high-K contraction. The tissue cyclic GMP content was not changed by nitroarginine or by removal of endothelial cells, but was elevated by stimulation with NA, ACh or SP by a factor of about 3, 5 or 11 times, respectively. These actions of ACh or SP were endothelium-dependent, and were inhibited by nitroarginine and remained unaltered by high-K solution. Thus, ACh and SP relax muscles indirectly by releasing endothelial factors, and the former by releasing mainly an endothelium-derived relaxing factor (EDRF), and the latter by releasing EDRF and other unidentified factors. As the relaxing actions of the latter factors are inhibited by high-K solution with no relation to the production of cyclic GMP, an involvement of hyperpolarizing factor, possibly EDHF, is suggested.

Acetylcholine↗

PubMed1996

Properties of junction potentials in gastric smooth muscle of the rat.

In isolated smooth muscles of the rat stomach, the properties of electrical responses of the membrane elicited by transmural nerve stimulation were studied. The smooth muscle membranes were quiescent in the fundus and spontaneously active with slow waves and often action potentials superimposed on top of the slow wave in the antrum and pylorus. The maximum membrane potentials were larger in the antrum (-50 to -55 mV) than in the fundus (-40 to -45 mV). Transmural nerve stimulation elicited an excitatory junction potential (e.j.p.) which was followed by an inhibitory junction potential (i.j.p.) in the fundus, and an i.j.p. alone in the antrum. The e.j.p. was inhibited by atropine, indicating that this potential was cholinergic in nature. The amplitude of the e.j.p. was increased by apamin or nitroarginine, and the latter was more potent than the former. The i.j.p. was inhibited by apamin or nitroarginine, and was resistant to adrenergic and cholinergic blocking agents, and therefore this potential was non-adrenergic non-cholinergic (NANC) in nature. The inhibitory actions of nitroarginine on the i.j.p. were antagonized by L-arginine, suggesting the involvement of nitric oxide (NO) in this junctional transmission. The results indicate that smooth muscles of the rat stomach receive cholinergic excitatory and NANC inhibitory nerves, and that endogenous NO may either be partly responsible for the generation of the i.j.p. or may modulate the junctional transmissions.

Animals↗

PubMed1995

Dual effects of trimebutine on electrical responses of gastric smooth muscles in the rat.

The effects of trimebutine on the electrical properties of smooth muscle membranes were studied in the isolated rat stomach, the objective being to elucidate the dual actions of this drug on gastric motility. Transmural nerve stimulation elicited a cholinergic excitatory junction potential (e.j.p.) and a nonadrenergic noncholinergic inhibitory junction potential (i.j.p.), and trimebutine inhibited the e.j.p. more than the i.j.p., with no significant change in the acetylcholine-induced depolarization. Trimebutine reduced the interval and, at high concentrations, the amplitude of slow waves. In enzymatically dispersed single cells, the Ca2+ current elicited by depolarization of the membrane was also inhibited by trimebutine. Thus, trimebutine increases slow wave frequency and inhibits cholinergic transmission and Ca2+ influx. The former would enhance while the latter two would depress gastric motility.

Animals↗

PubMed1993

Transgenic Drosophila carrying mammalian cytochrome P-4501A1: an application to toxicology testing.

Transgenic Drosophila which carry a canine cytochrome P-450 were established and evaluated for its value on toxicology testing. Dah1, a cDNA clone for the canine hepatic P-4501A1, was ligated between the promoter of Drosophila heat-shock protein gene and the terminator of actin 5C gene, and then microinjected into Drosophila embryos with a transposon P element. The injected DNA was stably integrated into chromosomal DNA in germ line cells. After heat-shock treatments, transgenic larvae produced mRNA and a detectable amount of protein that cross-reacted with antibodies against rat P-4501A1. The ability of transgenic flies to activate procarcinogens was monitored in the DNA repair test with the endpoint being preadult mortality of repair-defective mei-9a mei-41D5/Y males. Heat-shock treatments of larvae showed that the repair-defective males carrying the P-4501A1 transgene were killed by lower concentrations of 7,12-dimethylbenz[a]anthracene than their non-transgenic or non heat-shocked counterparts. Coadministration of alpha-naphthoflavone, one of the inhibitors of P-4501A subfamily, cancelled the enhanced mortality of transgenic males, supporting that 7,12-dimethylbenz[a]anthracene was metabolized to its genotoxic form by the expressed P-4501A1 enzyme. This study indicates a potential utility of transgenic Drosophila for screening mammalian promutagens and procarcinogens.

9,10-Dimethyl-1,2-benzanthracene↗