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J S Kelly

Publications and source records attributed to J S Kelly.

At least 37 records · Page 2Linked to original sources
PubMed1996

Glucocorticoids block protein kinase A inhibition of calcium-activated potassium channels.

Adrenal corticosteroids have well known and profound effects on neurons and neuroendocrine cells, but the underlying cellular mechanisms are poorly understood. The present study analyzed membrane currents and ACTH release in AtT20 mouse pituitary corticotrope tumor cells. Patch-clamp analysis revealed a significant and selective inhibition of calcium-activated (BK-type) potassium channels upon activation of protein kinase A by corticotropin-releasing factor or 8-chlorophenylthio-cAMP. The synthetic glucocorticoid dexamethasone had no effect on potassium currents evoked by depolarization but prevented the inhibitory effect of protein kinase A activators. The action of dexamethasone had the hallmarks of protein induction, i.e. a lag time and sensitivity to inhibitors of DNA transcription and mRNA translation. In parallel, the specific BK channel blocker iberiotoxin abolished early glucocorticoid inhibition of corticotropin-releasing factor-stimulated ACTH secretion. In summary, the present data show that glucocorticoid-induced proteins render BK-type channels resistant to inhibition by protein kinase A and that this action of the steroid is pivotal for its early inhibitory effect on the secretion of ACTH.

Adrenocorticotropic Hormone↗
PubMed1995

Only 'de novo' long-term depression (LTD) in the rat hippocampus in vitro is blocked by the same low concentration of FK506 that blocks LTD in the visual cortex.

It has been proposed that the long-term depression (LTD) seen following low frequency stimulation (LFS) in the rat hippocampus involves calcineurin. We have tested this by examining the effect of FK506, a macrolide which blocks calcineurin at nanomolar concentrations, on synaptic transmission in the rat hippocampal slice at a concentration of 1 microM which has been shown to block LTD in the visual cortex. The effect of FK506 on long-term potentiation (LTP) and spontaneous transmitter release was also studied. The magnitude of LTD induced by LFS was 16.7 +/- 2.4% in control which was not significantly different from the 22.3 +/- 3.0% seen in the same preparations after exposure to FK506 for 25-30 min. In contrast the magnitude of LTD induced 'de novo' in preparations exposed to FK506 was significantly reduced. FK506 had no significant effect on LTP, miniature EPSP frequency, miniature EPSP amplitude, resting membrane potential or input resistance. These results, therefore, support the hypothesis that calcineurin is involved in 'de novo' LTD but it appears that an event is triggered by LFS whereby FK506-insensitive LTD can subsequently be activated by a second episode of LFS.

Animals↗
PubMed1995

Adenosine 3':5'-cyclic monophosphate mediates a 5-hydroxytryptamine-induced response in neonatal rat motoneurones.

5-Hydroxytryptamine (5-HT) is present in nerve fibres descending from the brainstem Raphe nuclei to motoneurones and its release is thought to exert excitatory actions. 5-HT, applied from the outside, directly depolarizes spinal and cranial motoneurones in slices. This action of 5-HT is mediated, in part, by an inwardly rectifying cationic current, Ih. In cardiac cells, an equivalent current, if, has been shown to be directly activated by adenosine 3':5'-cyclic monophosphate (cAMP) applied to the inside of the patch membrane. By applying the whole-cell method to thin slices of brainstem and spinal cord, we have shown that intracellularly applied camp and extracellularly applied dibutyryl camp or forskolin mimics the inward current induced by 5-ht. The selective cAMP phosphodiesterase inhibitor, Ro 20-1724, clearly prolonged the 5-HT-induced current. Maximal doses of dibutyryl cAMP or forskolin occluded the 5-HT-induced current. The broad spectrum protein kinase inhibitors 1-(5-isoquinolinesulphonyl)-2-methlypiperazine (H-7) and N-[2-(methylamino)ethyl]-5-isoquinolinesulphonamide (H-8) had no effect on the currents induced by 5-HT and forskolin. From these results, we suggest that activation of 5-HT receptors on the motoneuronal membrane stimulates formation of intracellular cAMP, thereby inducing the inward current, possibly by a direct action on Ih.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1995

The temperature dependence of high-threshold calcium channel currents recorded from adult rat dorsal raphe neurones.

The temperature dependence of HVA calcium channel currents, using barium as the charge carrier, was studied in acutely isolated adult rat dorsal raphe neurones. Current amplitude was found to be highly sensitive with a Q10 of between 1.7 and 2.5. The most sensitive component of current is that that is activated from hyperpolarized holding potentials, and inactivates during a 200 msec test pulse. The least sensitive is the more sustained current elicited from depolarized potentials. Increases in temperature were also found to cause an irreversible shift in the current-voltage relationship in the hyperpolarizing direction. By far the most temperature-sensitive property was the activation time constant with an extraordinary Q10 of between 10 and 12. This was not significantly affected by holding potential, though the time constant itself is dependent on the test potential. Increases in temperature to 25 degrees C or above revealed a fast inactivating component, not seen at lower temperatures. These findings suggest that there are at least three components of HVA current in dorsal raphe neurones. In addition, the remarkably high Q10 for activation kinetics suggests that the processes underlying calcium channel current activation are multifaceted and complex. The following paper puts forward a new hypothesis which attempts to explain the way in which neurotransmitters modulate the activation kinetics of HVA calcium channel currents.

Animals↗
PubMed1995

The modulation of calcium channel currents recorded from adult rat dorsal raphe neurones by 5-HT1A receptor or direct G-protein activation.

The effect 5-HT1A receptor activation on the temperature dependence of HVA calcium channel currents has been studied in acutely isolated DR neurones, using barium as the charge carrier. 8-OH-DPAT caused a reduction in the temperature dependence of the peak current amplitude. However the most dramatic effect of 8-OH-DPAT was a large reduction in Q10 for the current activation rate. This also occurred with direct G-protein activation using GTP gamma S. In the presence of GTP gamma S, current activation became bi-exponential, rather than mono-exponential as in the control situation. The time constants of both components were significantly slower than the controls, and the Q10 for both components was significantly lower. GDP beta S had no effect on the temperature dependence or kinetics of activation of HVA current. Depolarizing prepulses applied prior to test pulses were able to reverse the action of 8-OH-DPAT on the Q10 of the activation rate. When prepulses were applied to cells containing GTP gamma S, the activation rate Q10 was similar to control values. We postulate that the highly significant reduction in activation rate Q10, seen with both 8-OH-DPAT and GTP gamma S, is as a result of a change in the mechanism underlying activation of HVA channels on depolarization. Contrary to previous models of calcium current modulation our results show that the mechanisms responsible for slowed activation by transmitters and facilitation of the residual current by depolarizing prepulses have little in common. We present a new model of transmitter modulation of HVA current, consistent with a mechanistic approach to channel subunit structure.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1995

The use of brain slices and dissociated neurones to explore the multiplicity of actions of 5-HT in the central nervous system.

The cellular actions of 5-hydroxytryptamine (5-HT) on adult and neonatal rat central neurones have been investigated in detail using a combination of in vitro slice and dissociated neurone preparations. Patch-clamp recordings from acutely dissociated adult rat dorsal raphe neurones confirm data obtained using conventional slice preparations that 5-HT activates an inwardly rectifying potassium channel through a 5-HT1A receptor leading to hyperpolarization of the cell. Single-channel recordings indicate that this pathway requires only the involvement of a pertussis toxin-sensitive G-protein. Adult rat facial motoneurones in conventional slices are depolarized by 5-HT through a combination of mechanisms, closure of potassium channels and enhancement of the hyperpolarization-activated, cationic current, IH. Distinct receptors appear to mediate these two actions. Both mechanisms are present in visually indentified neonatal rat facial motoneurones in thin brain slices. Whole-cell patch-clamp recordings show the action of 5-HT on IH to mediate a caesium-sensitive inward current which can be mimicked by the adenylate cyclase activator, forskolin. The experimental data illustrate how a range of complimentary in vitro electrophysiological techniques can be employed to unravel neurotransmitter mechanisms and pharmacology.

Animals↗
PubMed1995

Use of esophageal or precordial stethoscopes by anesthesia providers: are we listening to our patients?

STUDY OBJECTIVE: To ascertain current anesthesia utilization of esophageal and precordial stethoscopes in U.S. anesthesia training programs. DESIGN: Prospective, single-blind, incidence study. SETTING: Operating rooms of three tertiary care hospitals with major academic anesthesiology departments. SUBJECTS: Anesthesia faculty [MD and certified registered nurse-anesthetist (CRNA) staff] and anesthesia trainees (anesthesiology residents and student nurse-anesthetists). INTERVENTIONS: observe and record the placement (stethoscope device appropriately positioned) and utilization (stethoscope in place and connected to the ear piece of the anesthesia provider) of the esophageal or precordial stethoscope during general, regional, and monitored anesthesia care. MEASUREMENTS AND MAIN RESULTS: During 520 anesthetics, an esophageal stethoscope was inserted in 68% of subjects, a precordial stethoscope was positioned in 16%, and an anesthetic stethoscope was absent in 16% of cases. Utilization (stethoscope connected to earpiece) ranged from a low of 11% of cases to a high of 45%, depending on the institution. Overall, providers were listening via an anesthetic stethoscope in only 28% of anesthetics. CONCLUSIONS: Our data suggest infrequent utilization of esophageal and precordial stethoscopes in anesthesia training institutions. Thus, current anesthesia training may be fostering an environment where providers overlook a valuable minimally invasive, and cost-effective continuous monitor of patients' dynamic vital organ function.

Anesthesia Department, Hospital↗
PubMed1995

Validation therapy: a case against.

1. Evidence for the success of validation therapy remains largely anecdotal. 2. Validation therapy persists despite a lack of research-based support for its effectiveness, perhaps because it has been more of a commercial success than a scientific one. 3. The effectiveness or therapeutic value of any form of treatment should be assessed objectively before it is adopted and a widespread commitment be made to its use.

Aged↗
PubMed1994

Differential effects of competitive (CGS19755) and non-competitive (MK 801) NMDA receptor antagonists upon local cerebral blood flow and local cerebral glucose utilisation in the rat.

The effects of the selective non-competitive NMDA receptor antagonist dizocilpine (MK801) and the competitive NMDA receptor antagonist CGS19755 upon local blood flow (lCBF) and local glucose utilisation (lCGU) were examined in 81 neuroanatomically discrete regions of the conscious rat brain using the [14C]iodoantipyrine and [14C]2-deoxyglucose quantitative autoradiographic techniques, respectively. Animals received dizocilpine (0.3 mg/kg), CGS19755 (30 mg/kg) or saline vehicle (2 ml/kg) 10 min prior to the initiation of lCGU studies while blood flow determinations were performed in parallel groups of animals 20 min after drug administration. Dizocilpine significantly increased lCGU in 33 of the 81 regions measured (most notably in cortical and subcortical limbic structures and in the basal ganglia) while reducing glucose use in seven brain areas (frontoparietal and somatosensory cortex, and in areas subserving auditory function). In contrast, CGS19755 significantly reduced lCGU use in 39 of the 81 areas examined while increases were observed in only three areas (anterior piriform cortex, substantia nigra pars reticulata, and posterior thalamic nucleus). Following Dizocilpine administration, there was evidence of widespread (64 of the 81 areas studied) increases in lCBF, while blood flow was reduced in the inferior colliculus. Significant increases in lCBF were also noted in 26 brain areas of CGS19755-treated rats while in one area (flocculus) blood flow was reduced. In saline-treated rats there was a close correlation between lCBF and lCGU. Dizocilpine administration was associated with an increase in the overall lCBF:lCGU ratio from 1.56 ml/mumol (in saline-treated rats) to 2.34 ml/mumol. In some brain areas (CA1 subfield of the dorsal hippocampus, somatosensory cortex and nucleus accumbens) there was evidence of focal disturbances in flow-metabolism relationship. While a similar increase in the overall lCBF-lCGU use ratio was evident in CGS19755 treated animals, there was no evidence of focal uncoupling of the flow metabolism relationship in any of the 81 brain areas examined. These data show that whilst both competitive and non-competitive NMDA receptor antagonists increased cerebral tissue perfusion beyond that required to meet underlying metabolic demand, focal disturbances in the flow metabolism relationship were observed only in dizocilpine-treated rats.

Animals↗
PubMed1994

Endothelin-1 induced middle cerebral artery occlusion: pathological consequences and neuroprotective effects of MK801.

In the present study we utilise the potent vasoconstrictor properties of endothelin-1 (Et-1) in a new model of middle cerebral artery occlusion in the anaesthetized rat. We evaluate the reproducibility of the model and examine the neuroprotective efficacy of the potent anti-ischaemic agent, MK801. Adult male SD rats received MK801 (5 mg/kg, n = 7) or saline vehicle (n = 7) 30 mins prior to the microinjection of Et-1 (60 pmol in 3 microliters) via a 31-g cannula stereotaxically positioned 0.5 mm above the middle cerebral artery. Three days after the injection of Et-1, rats were perfusion fixed, the brain removed, cryostat sectioned and processed for histological staining. Sections at eight predetermined levels were examined by light microscopy and the volume of infarction calculated. Following administration of Et-1, saline-pretreated rats exhibited a pattern of ischaemic damage similar to that previously reported following permanent occlusion of the rat middle cerebral artery. This pattern was characterised by a large volume of infarction covering the dorsal and lateral neocortex (98 +/- 12 mm3) and striatum (32 +/- 3 mm3) ipsilateral to the insult. Power analysis predicted a group size of 7 would be required for a 50% reduction in ischaemic damage to be recorded as statistically significant at the 5% level. Pretreatment with MK801 reduced cortical tissue damage by 51% (P = 0.026) but did not significantly alter either the pattern or volume of infarction (33 +/- 4 mm3; P = 0.95) in the striatum.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthetics↗
PubMed1994

Selective enhancement of an A type potassium current by dexamethasone in a corticotroph cell line.

Perforated patch recording was used to examine the effect of the synthetic steroid dexamethasone on the whole cell potassium (K+) current, in the mouse corticotroph tumour cell line AtT20/D16-16. In 15 out of 52 control cells (29%) there was a rapidly-activating, rapidly-inactivating K+ current of the A type, the amplitude of which was strongly dependent on the holding potential in use prior to its activation by depolarising voltage pulses, and which was blocked by 1 mM 4-aminopyridine (4-AP, n = 5). The effect of dexamethasone (100 nM, 2 h, 37 degrees C) was that the A current increased in prevalence (24 out of 31 cells, 77%), lost its dependence on holding potential (over the range studied), and as a result became significantly larger than in controls, for certain voltage steps (peak A current density was 18.5 +/- 2.4 pA/pF (n = 12) for control cells and 26.3 +/- 3.9 pA/pF (n = 18) for dexamethasone treated cells, for a step to +30 mV from -60 mV, values are mean +/- SEM). All cells exhibited a slowly-activating, sustained K+ current, which was unaffected by changes in the holding potential, unaffected by 4-AP and consisted of at least 3 components: one blocked by 30 mM tetraethylammonium(TEA) or 100 nM charybdotoxin (CTX); a second blocked by 100 nM apamin; and a third not blocked by TEA, CTX, apamin, clofilium (100 nM) or niflumic acid (0.1 mM). Dexamethasone produced no change in the slowly-activating, sustained current nor in any of its individual components. The effect of dexamethasone on the A current was completely blocked by 0.1 mM puromycin, a protein synthesis blocker, while puromycin alone did not affect the size or frequency of the A current, nor alter the slowly-activating, sustained current. Secretion studies using 4-AP confirmed that the A current has a role in stimulated adrenocorticotrophic hormone (ACTH) secretion. In summary, AtT20 cells contain at least four types of K+ current: an A current and 3 currents contributing to the slowly-activating current. Selective enhancement of the A current by dexamethasone, shown here to require synthesis of new protein, is one of the mechanisms whereby glucocorticoids exert inhibitory control on ACTH secretion.

Adrenocorticotropic Hormone↗
PubMed1993

Characterisation of the binding of [3H]FR115427, a novel non-competitive NMDA receptor antagonist, to rat brain membranes.

The binding of [3H]FR115427 ([3H](+)-1-methyl-1-phenyl-1,2,3,4- tetrahydroisoquinoline) to rat cortical synaptosomal membranes was investigated. Binding was optimal at pH 7.4-8.0, and temperature had little effect on specific binding. Binding reached equilibrium within 30 min at 25 degrees C, and was reversible in the presence of excess unlabelled FR115427. [3H]FR115427 bound to a single population of non-interacting sites with an affinity of 45.4 +/- 3.9 nM, and a binding site density of 9.12 +/- 0.52 pmol/mg protein. The affinities of other N-methyl-D-aspartate (NMDA) receptor channel blockers for [3H]FR115427 binding sites were consistent with binding to a similar site to that occupied by dizocilpine. Binding was potentiated by L-glutamate and glycine with EC50 values of around 80 nM. In the presence of L-glutamate (10 microM), specific binding was increased 4-fold, whilst addition of glycine (10 microM) increased specific binding 2-fold. FR115427 exhibited marked stereoselectivity; (+)-FR115427 has 100-fold higher affinity than (-)-FR115427. This ligand may therefore be useful for the pharmacological investigation of the NMDA receptor ion channel.

Animals↗
PubMed1993

Neurochemical and electrophysiological studies on FR115427, a novel non-competitive NMDA receptor antagonist.

The pharmacological profile of FR115427 has been examined using ligand binding and electrophysiological techniques. Binding of [3H]dizocilpine in the presence of L-glutamate was inhibited by the (+) isomers of dizocilpine and FR115427. The corresponding (-) isomers were less active, and stereoselectivity was particularly marked in the case of FR115427. In contrast to dizocilpine, the affinity of FR115427 for [3H]dizocilpine binding sites was little affected by addition of either L-glutamate and/or glycine. In a cortical wedge preparation, FR115427 inhibited N-methyl-D-aspartate (NMDA)-induced responses in a non-competitive, use-dependent manner. Intracellularly recorded excitatory synaptic responses in hippocampal neurones were only partially inhibited by FR115427 thereby confirming a selective effect on the NMDA-mediated component of neuronal excitation induced by the endogenous neurotransmitter. The data suggest that FR115427 is a non-competitive, use-dependent NMDA receptor antagonist with more pronounced stereoselectivity and less marked use dependence than dizocilpine.

Action Potentials↗
PubMed1993

Binding of 5-HT1A receptor and 5-HT transporter ligands in rat cortex and hippocampus following cholinergic and serotonergic lesions.

The cellular localisation of 5-HT1A receptor and 5-HT transporter binding sites in the rat cortex and hippocampus has been examined. Lesions of either basal forebrain neurones or serotonergic neurones did not affect [3H]8-OH-DPAT binding, suggesting that 5-HT1A binding sites are not localised on cholinergic or serotonergic nerve terminals. The binding of the 5-HT transporter ligand, [3H]citalopram was unaffected by the cholinergic lesion whereas binding was reduced in both the hippocampus and cortex following serotonergic lesions. A reduction in binding site density rather than an alteration in affinity was responsible for this effect. While these data suggest that [3H]citalopram binding sites are located on serotonergic nerve terminals, the abolition of hippocampal binding sites contrasted with a 50% loss in cortical tissue.

5,7-Dihydroxytryptamine↗