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R A North

Publications and source records attributed to R A North.

At least 145 records · Page 8Linked to original sources
PubMed1991

Membrane properties and synaptic responses of rat striatal neurones in vitro.

1. A tissue slice containing a section of striatum was cut obliquely from rat brain so as to preserve adjacent cortex and pallidum. Intracellular recordings were made from 368 neurones, using either conventional or tight-seal configurations. 2. Two types of neurone were distinguished electrophysiologically. Principal cells (96%) had very negative resting potentials (-89 mV) and a low input resistance at the resting membrane potential (39 M omega): membrane conductance (10 nS at -65 mV) increased within tens of milliseconds after the onset of hyperpolarization (99 nS at -120 mV). Secondary cells (4%) had less negative resting potentials (-60 mV) and a higher input resistance (117 m omega at the resting potential): hyperpolarization caused an inward current to develop over hundreds of milliseconds that had the properties of H-current. 3. Most principal cells were activated antidromically by electrical stimulation of the globus pallidus or internal capsule. Intracellular labelling with biocytin showed that principal cells had a medium sized soma (10-18 microns), extensive dendritic trees densely studded with spines and, in some cases, a main axon which extended towards the globus pallidus. 4. Electrical stimulation of the corpus callosum or external capsule evoked a depolarizing postsynaptic potential. This synaptic potential was reversibly blocked by a combination of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) and DL-2-amino-5-phosphonovaleric acid (APV, 30 microM), but was unaffected by bicuculline (30 microM) and picrotoxin (100 microM). The underlying synaptic current had a fast component (time to peak about 4 ms), the amplitude of which was linearly related to membrane potential and which was blocked by CNQX; in CNQX the synaptic current had a slower component (time to peak about 10 ms) which showed voltage dependence typical of N-methyl-D-aspartate (NMDA) receptors. Both currents reversed at -5 mV. 5. Focal electrical stimulation within the striatum (100-300 microns from the site of intracellular recording) evoked a synaptic potential that was partially blocked (45-95%) by CNQX and APV: the remaining synaptic potential was blocked by bicuculline (30 microM). The bicuculline-sensitive synaptic current reversed at the chloride equilibrium potential. 6. The findings confirm that the majority of neostriatal neurones (principal cells, medium spiny neurones) project to the pallidum and receive synaptic inputs from cerebral cortex mediated by an excitatory amino acid acting through NMDA and non-NMDA receptors. These cells also receive synaptic inputs from intrinsic striatal neurones mediated by GABA.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1991

Stimulation by human chorionic gonadotropin of prostaglandin synthesis by early human placental tissue.

Successful establishment of pregnancy is dependent on inhibition of clotting and suppression of the maternal immune response at the feto-maternal interface. Early human placental production of prostacyclin (PGI2) and prostaglandin E2 (PGE2) may be important in this process. To examine the possible role of these PGs, we studied PGE and 6-keto-PGF1 alpha (stable metabolite of PGI2) synthesis in human placental (9-17 weeks gestation) organ cultures, and monolayer cultures of purified trophoblasts. PGE2 appeared to be the major protanoid formed. Other arachidonic acid metabolites identified in placental organ culture were 6-keto-PGF1 alpha, thromboxane B2, PGF2 alpha, leukotriene B4, 5(S)-hydroxyeicosatetraenoic acid (5-HETE), 12-HETE, and 15-HETE. The synthesis of PGE and 6-keto-PGF1 alpha altered with gestation and was maximal in the younger placentas. Arachidonic acid (33 microM) stimulated and indomethacin (28 microM) inhibited PG production. hCG, including physiological concentrations, stimulated PGE and 6-keto-PGF1 alpha synthesis in placental organ cultures. This effect was most striking in the 9-12 week placentas, compared to 15-17 week placentas. A similar hCG-induced stimulation of PGE production occurred in monolayer cultures of trophoblasts. The addition of TSH, FSH, and LH indicated that this effect was specific for hCG. These data suggest that hCG may have a biological role in the regulation of PG synthesis in early human placenta.

6-Ketoprostaglandin F1 alpha↗
PubMed1991

An in-depth investigation into a food poisoning outbreak.

On and shortly after the 6th May 1990, 16 people were affected by food poisoning in an old peoples' residential home, of whom two died. The vehicle of infection was identified as a baked Alaska contaminated by Salmonella enteritidis phage type (PT) 8 and, at an early stage of the investigation, the source was attributed to a single infected egg. A separate investigation by the author, however, revealed that the baked Alaska meringue had been dispensed from an inadequately cleaned piping bag which had been recovered from the kitchen a month after the outbreak. A pure, profuse culture of S. enteritidis PT8 was isolated from it. At least one secondary case may have been attributable to food made with this bag. Ministry of Agriculture Investigations of the flocks suspected of producing the eggs used for the baked Alaska demonstrated an absence of S. enteritidis. On this basis, the author considered a more likely cause of the outbreak to be the piping bag, contaminated from source or sources unknown within the kitchen. Furthermore, the possibility of human carrier transmission cannot be wholly ruled out. The incident underlines the dangers of jumping to conclusions at the outset of food poisoning investigations and emphasises that hypotheses formulated on sources of contamination must be properly tested, the absence of which, in this instance, led the investigators to unwarranted conclusions as to the cause of the outbreak.

Aged↗
PubMed1991

Baclofen and adenosine inhibit synaptic potentials mediated by gamma-aminobutyric acid and glutamate release in rat nucleus accumbens.

Intracellular recordings were made from rat nucleus accumbens neurons in a tissue slice in vitro; postsynaptic potentials (p.s.p.) were evoked by focal electrical stimulation of the slice surface. P.s.p. were partially blocked by bicuculline (30 microM), partially blocked by a combination of 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX, 10 microM) and DL-2-amino-5-phosphonovaleric acid (APV, 30 microM) and completely blocked when all three antagonists were applied together. Both the gamma-aminobutyric acid (GABA)-mediated p.s.p. (in CNQX and APV) and the glutamate-mediated p.s.p. (in bicuculline) were inhibited by baclofen (10-300 microM), adenosine (10-300 microM) and N6-(2-phenylisopropyl)adenosine (0.1-3 microM). Theophylline competitively antagonized the action of adenosine with an apparent dissociation equilibrium constant of about 15 microM. Baclofen, adenosine and N6-(2-phenylisopropyl)adenosine caused small (less than 10 mV) hyperpolarizations; voltage clamp experiments indicated that this resulted from an outward potassium current. It is concluded that activation of GABAB receptors and adenosine A1 receptors inhibits the release of glutamate and GABA at synapses in the nucleus accumbens.

2-Amino-5-phosphonovalerate↗
PubMed1991

Adenosine A1 and A2 receptors mediate presynaptic inhibition and postsynaptic excitation in guinea pig submucosal neurons.

Intracellular recordings were made from guinea pig submucosal neurons in vitro. Adenosine, 2-[p-(carboxyethyl)phenylethylamino]-5'-N- ethylcarboxamidoadenosine (CGS21680), 2-chloroadenosine (CADO), 5'-N-ethylcarboxamidoadenosine (NECA), R(-)-N6-(2-phenylisopropyl)adenosine (R-PIA), N6-cyclohexyladenosine (CHA) and 1-deaza-2-chloro-N6-cyclopentyladenosine (DCCPA) were applied by adding them to the superfusion solution. Adenosine (30 nM to 30 microM) depolarized S-type neurons neurons and this was mimicked by analogs with potency order: CGS21680 = NECA greater than R-PIA greater than CADO greater than adenosine greater than CHA much greater than DCCPA. 8-Cyclopentyltheophylline (CPT) blocked the depolarizing action of CADO or R-PIA; this antagonism was surmountable and the dissociation equilibrium constant (KD) estimated by the Schild method was 295 nM. Synaptic potentials were evoked by focal stimulation of nerve strands running between submucosal ganglia. The nicotinic excitatory postsynaptic potential was reduced by adenosine and analogs with potency order CHA = R-PIA greater than DCCPA = NECA = CADO greater than adenosine much greater than CGS21680; CPT competitively antagonized this effect of CADO or CHA with a KD of 13 nM. The noradrenergic inhibitory postsynaptic potential was also reduced; the potency order was R-PIA greater than CHA = CADO = NECA = DCCPA greater than adenosine much greater than CGS21680; the KD of CPT as an antagonist was 7 nM. It is concluded that adenosine directly depolarizes submucosal neurons by acting at an A2 receptor, and that it inhibits the release of acetylcholine (from intramural nerves) and noradrenaline (from sympathetic nerves) by acting at a presynaptic A1 receptor.

Acetylcholine↗
PubMed1991

Identification of amino acid residues involved in dendrotoxin block of rat voltage-dependent potassium channels.

alpha-Dendrotoxin (DTX) is a 60-amino acid peptide belonging to the family of mamba snake neurotoxins; it is a potent blocker of some but not all voltage-gated potassium currents. Potassium currents recorded from oocytes injected with cloned potassium channel RNAs also vary in sensitivity to DTX. Expression of channels that were chimeras of the DTX-sensitive channel RBK2 and the DTX-insensitive channel RGK5 showed that the putative extracellular loop between transmembrane domains S5 and S6 contributes strongly to DTX sensitivity. Mutation of two residues (Ala352Glu353) in this region of RBK1 to conform to those at equivalent positions in RGK5 (Pro374Ser375) reduced the potency of DTX about 70-fold, and the substitution of Tyr379 in RBK1 by its counterpart in RGK5 (His401) caused an additional 2.5-fold decrease in sensitivity. Converse substitutions in RGK5 significantly increased sensitivity to DTX. The results suggest that these residues contribute significantly to the channel-toxin interaction, providing further evidence that the S5-S6 loop lies at or near the external mouth of the channel, where DTX binding leads to channel occlusion. They offer a molecular explanation for the differences in DTX sensitivity observed among native potassium channels.

Amino Acid Sequence↗
PubMed1990

MK-801 blocks nicotinic depolarizations of guinea pig myenteric neurons.

Intracellular recordings were made from guinea-pig myenteric neurons. Acetylcholine (ACh), applied by iontophoresis, produced a depolarization that was blocked by hexamethonium (EC50 3.2 microM) but not by scopolamine (1 microM). MK-801 (0.3-30 microM), a non-competitive antagonist at N-methyl-D-aspartate receptors, also blocked the nicotinic depolarization (EC50 = 4.5 microM). Voltage clamp measurements of membrane current showed that MK-801 (1 microM) and hexamethonium (1 microM) both produced a greater inhibition of ACh-induced inward currents at -100 mV than at -40 mV. MK-801 (10 microM) did not change the depolarization evoked by 5-HT (acting at 5-HT3 receptors) in the same neurons in which it reduced the nicotinic response by more than 70%. It is concluded that MK-801 can act as a non-competitive antagonist of nicotinic responses in myenteric neurons.

Acetylcholine↗
PubMed1990

Heteropolymeric potassium channels expressed in Xenopus oocytes from cloned subunits.

Voltage-dependent potassium currents were measured in Xenopus oocytes previously injected with RNAs generated in vitro from each of three cloned cDNAs (RBK1, RBK2, and RGK5). The currents differed in their sensitivities to blockade by tetraethylammonium (TEA; respective KDs 0.3, greater than 100, and 10 mM) and in their inactivation during a depolarizing pulse. Injections of RNA combinations (RBK1/RBK2 and RBK1/RGK5) caused currents that had TEA sensitivities different from those expected from the sum, in any proportion, of the two native channels. It is concluded that novel potassium channels are formed by the oocytes injected with two RNAs, presumably by heteropolymerization of subunits; such heteropolymerization would contribute functional diversity to voltage-dependent potassium channels in addition to that provided by a large gene family.

Animals↗
PubMed1990

Actions of cocaine on rat dopaminergic neurones in vitro.

1. Intracellular recordings were made from neurones in vitro in the rat substantia nigra zona compacta and ventral tegmental area; the neurones had the characteristic properties of dopamine-containing cells, and fired action potentials spontaneously. 2. Cocaine (1-10 microM) inhibited spontaneous firing, hyperpolarized the membrane and (in neurones voltage-clamped at -60 mV) caused an outward membrane current; the minimally effective concentration was 1 microM. These effects were blocked by sulpiride (30 nM-1 microM). 3. Dopamine (3-100 microM) also inhibited firing, hyperpolarized and caused an outward current. These effects of dopamine were potentiated about five fold by cocaine (10 microM). 4. It is concluded that cocaine (1-10 microM) inhibits the firing and hyperpolarizes substantia nigra zona compacta neurones in vitro by blocking the uptake of dopamine which the cells continuously release.

Action Potentials↗
PubMed1990

Actions of cocaine on rat nucleus accumbens neurones in vitro.

1. Intracellular recordings were made from 103 neurones of the rat nucleus accumbens in vitro. 2. Dopamine (3-100 microM; in sulpiride, 1 microM) hyperpolarized neurones (79%) by acting at D1 receptors: dopamine (3-100 microM; in SCH23390, 1 microM) depolarized neurones (55%) by acting at D2 receptors. 5-Hydroxytryptamine (1-100 microM) depolarized 86% neurones. 3. Both actions of dopamine as well as the effect of 5-hydroxytryptamine were potentiated by cocaine (0.3-30 microM), which had no effect of its own on membrane potential. 4. Dose-ratio was computed as [(concentration of agonist causing a 4 mV potential change in cocaine)/(concentration of agonist causing a 4 mV potential change without cocaine)]. Cocaine (1-30 microM) caused the same dose-ratio whether dopamine depolarizations (D2) or hyperpolarizations (D1) were measured; the dose-ratio ranged from 2 (1 microM) to 50 (30 microM). 5. Responses to 5-hydroxytryptamine were increased more than responses to dopamine; cocaine 1 microM gave a dose-ratio of 13.4 and at 30 microM gave a dose-ratio of 118. 6. It is concluded that cocaine acts to inhibit the uptake of dopamine and 5-hydroxytryptamine in slices of rat nucleus accumbens; lower concentrations of cocaine (0.3 to 1 microM) are particularly effective in potentiating the action of 5-hydroxytryptamine.

Animals↗
PubMed1990

Muscarine reduces inwardly rectifying potassium conductance in rat nucleus accumbens neurones.

1. Intracellular recordings were made from neurones in the nucleus accumbens in slices from the rat brain maintained in vitro. 2. Muscarine (1-100 microM) depolarized 101 of 107 neurones; this was associated with an increase in the input resistance. The potential change reversed polarity with conditioning hyperpolarization and the reversal potential was linearly related to the logarithm of the extracellular potassium concentration. 3. The depolarization caused by muscarine was not changed by tetrodotoxin (1 microM) or by a solution that contained lower levels of calcium (0.24 instead of 2.4 mM), higher levels of magnesium (5 instead of 1.2 mM) and cobalt (2 mM). 4. Muscarine caused an inward current and a decrease in slope conductance when applied to neurones voltage clamped near their resting potential (-82 mV). The current caused by muscarine reversed polarity at the potassium equilibrium potential. The current-voltage relation of the neurones between -60 and -120 mV was well fitted by assuming a voltage-independent potassium conductance and an inward rectifier potassium conductance; muscarine reduced predominantly the inward rectifier conductance. 5. Phorbol-12,13-diacetate (3 microM) and 5-hydroxytryptamine mimicked the action of muscarine. The inward currents caused by muscarine or 5-hydroxytryptamine were occluded by the inward current evoked by the phorbol ester. 6. The depolarization caused by muscarine was competitively antagonized by pirenzepine; the dissociation constant of 11 nM suggested involvement of the M1 receptor. 7. It is concluded that muscarine acts at M1 receptors to reduce the membrane potassium conductance and that activation of protein kinase C may be an intermediate step.

Action Potentials↗
PubMed1990

Inhibition of calcium currents by noradrenaline, somatostatin and opioids in guinea-pig submucosal neurones.

1. Whole-cell recordings were made from submucosal neurones acutely dissociated from guinea-pigs. The actions of noradrenaline, somatostatin and [Met5]enkephalin on currents carried by calcium ions were studied. 2. On depolarization from a holding potential of -70 mV, an inward current activated at -40 mV, reached its peak amplitude at 10 mV and reversed to outward at 72 mV (with external calcium of 5 mM and internal caesium of 160 mM). 3. Cadmium, nickel and cobalt reversibly blocked the calcium current; concentrations causing 50% block were 2.5, 500 and 2000 microM respectively. The calcium current (holding at -70 or -30 mV) was reversibly blocked by omega-conotoxin (100 nM), and unaffected by Bay K 8644 (0.1-10 microM) and nifedipine (1 microM). Cadmium caused an outward shift in holding current at -30 mV, implying that there was a persistent inward calcium current at this potential. 4. Noradrenaline, somatostatin and [Met5]enkephalin decreased the calcium current. The maximal inhibition observed with any one agonist, or with a combination of two agonists, did not exceed 50%; concentrations giving half-maximal inhibition were 5.5 microM for noradrenaline, 4 nM for somatostatin and 1 microM for [Met5]enkephalin. The inhibition was independent of membrane potential. All three agonists also reduced the persistent calcium current at -30 mV. 5. Inhibition of the calcium current by noradrenaline occurred with a latency of not less than 175 ms; cadmium applied by the same method depressed the current within 5-45 ms. 6. Experiments with selective agonists and antagonists indicated that the receptor types involved in calcium current inhibition were alpha 2-adrenoceptors and delta-opioid receptors. Somatostatin acted at a distinct receptor. 7. Calcium currents were also inhibited by intracellular dialysis with guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S). Agonists were ineffective in cells pre-treated with pertussis toxin, but their action was restored when purified GTP-binding proteins (Go or Gi) were included in the intracellular recording solution. 8. It is concluded that noradrenaline, somatostatin and [Met5]enkephalin act at their respective receptors on guinea-pig submucosal neurones to inhibit a voltage-dependent calcium current. Activation of the same receptors also increases a potassium conductance in these cells: in both cases a pertussis-sensitive G protein is involved.

Action Potentials↗
PubMed1990

Cation current activated by hyperpolarization (IH) in guinea pig enteric neurons.

Intracellular microelectrode and whole cell patch-clamp recordings were made from guinea pig enteric neurons in vitro. In most myenteric AH neurons (but not S neurons), step hyperpolarizations from the resting potential evoked an inward current (IH). IH peaked in approximately 400 ms (at 36 degrees C), was fully activated at -100 mV, and did not inactivate during 10 s. IH was associated with a conductance increase and reversed polarity at about -40 mV (by extrapolation). IH was reduced in low-sodium concentrations and increased when the concentration of extracellular potassium ions was increased. Cesium (2 mM) blocked IH in a voltage-dependent manner; this led to an increase in the amplitude of the spike after-hyperpolarization. Cobalt (2-4 mM) or barium (0.01-1 mM) did not alter IH. Only 12% of submucous plexus neurons showed IH and this subgroup included both S and AH neurons. In myenteric AH neurons, IH would act in opposition to the persistent calcium-activated potassium current and thus stabilize the resting potential.

Action Potentials↗
PubMed1990

Cation current activated by hyperpolarization in a subset of rat nucleus accumbens neurons.

1. Intracellular recordings were made from neurons in slices of rat nucleus accumbens in vitro. Membrane currents were measured in the potential range -60 to -120 mV with the use of a single-electrode, voltage-clamp amplifier. 2. A minority of neurons (28/285) was identified that had resting membrane potentials almost 20 mV less negative than the majority of the cells. These cells, but not the majority, had an inward current that activated slowly when the cells were hyperpolarized from -60 to -120 mV. The time constant of activation was approximately 3 s at -70 mV and 100 ms at -120 mV. 3. This inward current was completely blocked by external cesium (2 mM) but unaffected by barium. The current was reduced in solutions containing low-sodium concentration and increased in solutions with high-potassium concentration; its reversal potential was estimated to be -36 mV. 4. It is concluded that two types of neuron can be distinguished in the rat nucleus accumbens on the basis of the presence or absence of a cation current activated by hyperpolarization. This current (IH, also called If and IQ) causes the neurons to have less-polarized resting potentials than the majority of nucleus accumbens neurons.

Animals↗
PubMed1990

Potassium conductance increased by noradrenaline, opioids, somatostatin, and G-proteins: whole-cell recording from guinea pig submucous neurons.

Agonists at alpha 2-adrenoceptors, delta-opioid receptors, and somatostatin receptors were applied to dissociated guinea pig submucous plexus neurons; whole-cell recordings of membrane current showed that they increased the membrane potassium conductance. The conductance affected showed inward rectification, being described by Gag(max)/[1 + exp((V - V0.5)/k)] where V0.5 was about -65 mV and Gag(max) was about 10 nS. The agonists were ineffective when the potassium conductance of the neurons had first been increased by intracellular dialysis with purified guanosine 5'-triphosphate (GTP)-binding proteins (Gi or Go). Agonist actions were prevented by pertussis toxin, applied intracellularly (10-100 ng/ml for several minutes) or extracellularly 1-10 micrograms/ml for 1 hr); in the latter case, the agonist responses were reconstituted by intracellular dialysis with GTP-binding proteins.

Adrenergic alpha-Agonists↗
PubMed1990

Muscarine depolarizes rat substantia nigra zona compacta and ventral tegmental neurons in vitro through M1-like receptors.

Intracellular recordings were made from presumed dopamine-containing neurons in slices of rat mesencephalon. Muscarine (3-100 microM) increased the rate of spontaneous action potentials; it also caused a membrane depolarization and, in voltage-clamp, an inward current. Concentration-effect curves to muscarine were shifted rightwards by pirenzepine (0.03-1 microM) with an estimated KD of 14 nM. The inward current caused by muscarine was voltage-dependent. Between about -50 and (-)-65 mV it was associated with a decrease in membrane conductance, but between -70 and -110 mV it was unaccompanied by any change in membrane conductance. Muscarine was without effect on the action potential afterhyperpolarization, or on a slowly developing inward current evoked by step hyperpolarizations of up to 20 mV from -45 mV. Muscarinic depolarizations or inward currents were reduced reversibly or abolished by a low calcium (0.25 mM)/high magnesium (10 mM) solution. It is concluded that muscarinic excitation of dopaminergic neurons is mediated by M1-like receptors.

Animals↗
PubMed1989

5-HT3 receptors are membrane ion channels.

The neurohormone 5-hydroxytryptamine (5HT or serotonin) exerts its effects by binding to several distinct receptors. One of these is the M-receptor of Gaddum and Picarelli, now called the 5-HT3 receptor, through which 5-HT acts to excite enteric neurons. Ligand-binding and functional studies have shown that the 5-HT3 receptor is widely distributed in peripheral and central nervous tissue and evidence suggests that the receptor might incorporate an ion channel permeable to cations. We now report the first recordings of currents through single ion channels activated by 5-HT3 receptors, in excised (outside-out) membrane patches from neurons of the guinea pig submucous plexus. Whereas application of acetylcholine activated predominantly a 40-pS channel, 5-HT caused unitary currents apparently through two channels of conductances of 15 and 9 pS, which were reversibly blocked by antagonists of the 5-HT3 receptor. Receptors for amine neurotransmitters, including 5-HT1 and 5-HT2, have previously been thought to transduce their effects through GTP-binding proteins: the direct demonstration that 5-HT3 receptors are ligand-gated ion channels implies a role for 5-HT, and perhaps other amines, as a 'fast' synaptic transmitter.

Acetylcholine↗