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Quantal analysis of synaptic transmission in CA1 pyramidal cells of the rat hippocampusIsaac, John Timothy Roger January 1993 (has links)
No description available.
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Single-channel recordings of potassium channels from guinea-pig inner hair cellsAppenrodt, Peter January 1997 (has links)
No description available.
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Patch-clamp studies of single type-1 Ins(1,4,5)P3 receptor channelsDargan, Sheila Louise January 2001 (has links)
No description available.
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The Structural Basis for Ligand Recognition by Mouse Odorant ReceptorsRepicky, Sarah Elizabeth 22 April 2008 (has links)
Mammalian odorant receptors (ORs) are Class I G-protein coupled receptors (GPCRs) located within the nasal epithelium. Odorant receptors interact with Galpha olfactory, a Galpha S type G-protein. Activated Galpha olfactory stimulates adenylate cyclase and the resulting increase in cAMP concentration opens cyclic nucleotide gated channels allowing Ca2+ to enter the cell. The increased Ca2+ then activates a Ca2+ activated Cl- channel which further depolarizes the cell. This depolarization initiates an action potential that reaches the axon of the olfactory sensory neuron located in the main olfactory bulb. Information from the main olfactory bulb is then transmitted to higher regions of the brain. Olfactory information is initially coded through the interaction of odorant molecules with hundreds of distinct ORs, but difficulty in exogenous expression of odorant receptors has delayed the identification of ligands for individual ORs. However, expression of mouse odorant receptors in Xenopus laevis oocytes allows for a systematic screening for potential ligands, as well as for efficient study of the structure-function relationship of the receptors and their ligands. My screening of odorant receptors using Xenopus oocytes included the coexpression of a signal transduction system and the use of robotic two-electrode voltage clamp electrophysiology. In this study, I investigated the structural basis for ligand recognition in mouse odorant receptors. First, I expanded the molecular receptor ranges of seven Class I odorant receptors. By use of a high throughput assay, I was able to expand upon current knowledge in the field for the mouse odorant receptors 23-1, 31-4, 32-11, 40-4, 42-1, 42-2 and 42-3. I then examined one receptor (MOR23-1) in more detail. I used the substituted cysteine accessibility method to identify residues within transmembrane domain five of this receptor that are accessible from the extracellular space. These residues may line the ligand binding site or the ligand access pathway. Conventional mutations of A205 caused little alteration in the molecular receptive range of the receptor, suggesting that this residue may not play a significant role in ligand interaction within the binding pocket. Mutagenesis of G111, a residue within transmembrane domain three caused significant shifts in the molecular receptive range of the receptor, but the location of this residue within the binding pocket could not be confirmed by the substituted cysteine method. Previous reports had suggested significant similarity between the molecular receptive ranges of the seven mouse odorant receptors that I used in my research. By expanding upon the known aliphatic ligands for each receptor identified new ligands for each receptor, I was able to show that the molecular receptive ranges of these receptors are in fact distinct. The experimental identification of residues located within the binding pocket on transmembrane five of mouse odorant receptor 23-1 provides an improved understanding of ligand recognition by this receptor class and will aid in better computer modeling of these receptors. This increased accuracy of the computer models of these basic Class I GPCRs may aid in future drug discoveries. Since GPCRs constitute a significant fraction of current drug targets, understanding the mechanism of ligand interactions with mouse odorant receptors may aid in the development of more efficacious compounds in the treatment of many common ailments.
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The Effect of Clamp Support on the Pipe to T(0,1) Guided Wave and Its SimulationKuo, Chun-hung 31 August 2007 (has links)
In this study, to discuss the effect of the boundary between pipeline and clamp support changed by different pressures to the propagation of guided wave in the pipeline is the main idea. In addition, the author simulates the wave propagation situation by using finite element method. In this work, T(0,1) torsional mode was used to discuss when adding different pressures to the clamp support, the change of its reflection coefficients in different frequencies by the axial symmetric property propagates in the cylinder pipe. In the simulation, we take the ¡§fictitious layer¡¨ was used to describe the situation between the clamp support and pipeline when adding different pressures. Moreover, the stiffness normal to the fictitious layer and the stiffness parallel to the fictitious layer were taken as material parameters to achieve the situation between clamp support and pipeline. According to experimental results, when the torque increases, the reflection coefficients will decrease with increasing frequency. The reflection coefficients are about 0.08 to 0.02. By the result of experiment and simulation, one can know that when adding torque on the clamp support increases, the reflection coefficient will decrease. In addition, the author also prove that if we change the stiffness parallel to the fictitious layer material factor, then the T(0,1) guided wave will be more sensitive by its action of particle motion.
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Structural examination of voltage gated potassium channels by voltage clamp fluorometryVaid, Moninder 05 1900 (has links)
Voltage clamp fluorometry (VCF) was first developed in the mid 1990s by Isacoff and his colleagues. In this approach fluorophores are attached to substituted cysteine residues that are engineered by site-directed mutagenesis. Changes in the dielectric environment of the fluorophore report local transitions that are associated with electrically-related and electrically-silent transitions. VCF provides a powerful technique to observe real time reports of ion channel gating conformations. It has proven to be a useful technique because it adds insight that is not available using other techniques. X-ray crystallography studies give a predominantly static picture of the channel, while patch clamping of channels gives information only about residues that effect ionic current flow. Similarly, gating current provides insight only about residues that are charged and move across the membrane electric field.
In this thesis we examined the structural rearrangements of the Shaker channel and the effect of 4-AP on channel gating. We also examined for the first time the structural rearrangements of the Kv1.5 gating and the how the channel responds to depolarization pulses. This work is instrumental in the examination of the potassium channel gating.
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Cross-talk between nicotinic acetylcholine (nAChR) and serotonin (5HT3R) receptors in sympathetic neurons2013 September 1900 (has links)
Serotoninergic type 3 receptors (5HT3Rs) are members of the Cys-loop family of ligand-gated ion channels (LGIC), which includes nicotinic ACh, glycine, GABA-A and GABA-C receptors. All members of this family are widely expressed in the central and peripheral nervous systems, where they mostly participate in fast synaptic transmission. Activation of 5HT3Rs on vagal sensory nerve endings affect respiration, circulation, emesis and nociception; and in the central nervous system they are implicated in anxiety, depression, and drug dependence. In contrast, the function of 5HT3Rs in sympathetic neurons has not been fully determined. We discovered that 5HT3Rs interact with nicotinic acetylcholine receptors (nAChRs), the main drivers of the fast cholinergic autonomic synapse, through cross-talk mechanisms. We examined cross-talk by the patch-clamp technique on cultured mouse superior cervical ganglia (SCG) neurons. Co-stimulation of 5HT3Rs and nAChRs resulted in the generation of a combined current that was smaller than arithmetically predicted if the receptors did not interact with one another. This interaction, which we quantified as mean peak amplitude and mean ionic charge, was dependent on activation of 5HT3Rs and nAChRs, and independent of metabotropic receptors, Ca2+ entry and Ca2+ second messenger pathways, and of the direct action of 5HT on nAChRs. Preliminary data using an antibody targeted to the M3-M4 linker region of the 5HT3A subunit revealed that 5HT3Rs and nAChRs possibly cross-talk through physical interactions. These results revealed a potential role of the 5HT3R in the regulation of sympathetic synaptic transmission through cross-talk inhibition of nAChRs.
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Effects of Alismatis Rhizoma's Extract on Short ¡V Circuit Current and Conductance across Frog Skin EpitheliumPei, Jui-fa 30 May 2004 (has links)
In traditional Chinese herb medicine, Alismatis rhizoma has been used in treating edema, inflammation and increasing urine flow. Mechanism of Alismatis rhizoma¡¦s effect on these functions has not been elucidated. Since diuresis has been considered closely related to the reabsorption of sodium ion via the epithelium of tubule and collecting duct in kidney, we suspected that Alismatis rhizoma may influence transportation of salt and water.
The measurement of short circuit current ( Isc ) has been used widely to estimate the ion transportation between mucosal and serosal side of epithelium. In the present experiment, we used the voltage ¡V clamp technique to demonstrate the effect of Alismatis rhizoma¡¦s extracts ( ARE ) on Isc and sodium ion conductance in frog skin. Our result showed that in control stage, the potential difference ( PD ) of frog skin is 64.81 ¡Ó 2.44 mV, the Isc is 59.82 ¡Ó 3.58 £gA / cm2 and the conductance is 1.09 ¡Ó 0.18 £gA / cm2 / mV.
After ARE was applied to mucosal side of the frog skin, its Isc decrease from 62.63 ¡Ó 5.31 £gA / cm2 to 47.92 ¡Ó 5.41 £gA / cm2, which could further go down to 3.36 ¡Ó 1.06 £gA/cm2 by adding amiloride. Treating serosal side of frog skin with ARE decreased approximately 10% of its Isc. No apparent changes in conductance were observed by adding ARE to mucosal ( 0.98 ¡Ó 0.24 £gA / cm2 / mV ) or serosal side ( 0.96 ¡Ó 0.23 £gA / cm2 / mV ).
Adding L-arginine ( the precursor of nitric oxide ) to the serosal side of the skin epithelium elevated the Isc for 17.10 ¡Ó 9.30 £gA/cm2. This effect can be inhibited by applying ARE or NG-L-nitro-arginine methyl ester ( L-NAME, NO synthase inhibitor ) before application of L-arginine.
In summary, Alismatis rhizoma could affect Isc on both mucosal and serosal sides of the frog skin. Its effect on lowering Isc was more obvious when applied to the mucosal side than to the serosal side. The ARE may exert its effect on mucosal side by affecting amiloride¡Vsensitive sodium channel and on serosal side by affecting the NO signal transduction pathway.
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Properties of Action Potential Waveform-Evoked L-type Calcium Currents in Pituitary GH3 CellsLee, Chuan-Te 20 June 2002 (has links)
The purpose of this study was to examine the time course and kinetics of L-type Ca2+ current (ICa,L) from pituitary GH3 cells in response to various action potential (AP) waveforms using the whole-cell configuration of the patch-clamp technique. The major findings in this study are:
1. ICa,L evoked during the AP waveform exhibited an early and a late component. The early component of ICa,L occurred on the rising phase of the AP, whereas the late component coincided with the falling phase.
2. A prolongation of the falling phase of APs led to an increase in Ca2+ charge carried by ICa,L, although the amplitude of the late ICa,L was reduced.
3. When the peak voltage of AP waveforms was prolonged without changing the rising and falling phases, the amplitude of the late components was significantly increased.
4. ICa,L was also found to inactivate during a train of AP waveforms. When Ba2+ ions were used as the charge carriers, current inactivation during a train of APs was decreased.
5. The amplitude of ICa,L evoked by the AP templates with irregular bursting pattern was inactivated.
6. When spontaneous APs with the depolarizing potentials were replayed to GH3 cells, Ca2+ entry was not only spread over the entire AP, but also occurred during the interspike voltage trajectory.
7. When cells were exposed to thyrotropin releasing hormone (TRH; 10
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CONTRIBUTIONS OF EAG PROTEIN TO NEURONAL EXCITABILITY IN IDENTIFIED THORACIC MOTONEURONS OF DROSOPHILASrinivasan, Subhashini January 2010 (has links)
Diversity in the expression of ion channel proteins among neurons allows a wide range of excitability, growth and functional regulation. Ether-a-go-go (EAG), a member of the voltage-gated K+ channels, was characterized by spontaneous firing in nerve terminals and enhanced neurotransmitter release. In situ whole-cell patch-clamp recordings performed from the somata of Drosophila larval thoracic aCC motoneurons revealed spontaneous spike-like events in eag mutants. Spontaneous events were absent in wild type motoneurons. Spikes evoked by somatic current injection in to the cell body were not altered and comparable to wild type. Spontaneous spike-like events could be due to increased synaptic drive or altered intrinsic excitability of the motoneuron. Reduction of EAG function with selective expression of eag double stranded RNAi transgene in motoneurons only did not cause spontaneous spike-like events or alter evoked firing. This suggests increased synaptic drive contributes to spontaneous events.Both transient and sustained voltage-activated K+ currents, each with Ca++-sensitive (IA(Ca) and IK(Ca)) and Ca++ -insensitive components (IA and IK), were isolated in thoracic aCC motoneurons. In wild type motoneurons, IA was larger than IA(Ca). Conversely, IK(Ca) was larger than IK. Both eag mutants and eag RNAi expression resulted in a decrease in IA , IK and a slow sustained K+ current. Further, EAG and Shal demonstrate a potential functional interaction and contribute to IA. The voltage sensitivity for inactivation was reduced in Shal only and EAG-Shal double knock down compared to controls and EAG only knock down. In addition, a Ca++ sensitive EAG dependent K+ current was blocked by cAMP. Thus, both voltage-dependent and modulatory functions of EAG influence excitability in motoneurons.Firing properties and K+ currents distinguish aCC motoneurons in thoracic segments, T1 and T3. T3aCC had a shorter delay to spike, higher input resistance and were more easily recruited than T1aCC. T1aCC had a larger IA than T3aCC, but comparable IA(Ca). IK(Ca) was larger in T3aCC compared to T1aCC. These differences reflect cell-specific ion channel distribution that could contribute to patterned segmental motor output.
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