scholarly journals Modifications of single acetylcholine-activated channels in BC3H-1 cells. Effects of trimethyloxonium and pH.

1990 ◽  
Vol 96 (1) ◽  
pp. 1-22 ◽  
Author(s):  
P A Pappone ◽  
G L Barchfeld

We have examined the effects of chemical modification with trimethyloxonium (TMO) and changes in external pH on the properties of acetylcholine (ACh)-activated channels in BC3H-1 cells, a clonal muscle cell line. TMO reacts covalently and specifically with carboxylic acid moieties in proteins to convert them to neutral methyl esters. In BC3H-1 cells TMO modification reduces the whole-cell response to ACh measured at negative membrane potentials by approximately 60%. G omega seal patch-clamp recordings of single ACh channel currents showed that the reduction in ACh sensitivity is due to alterations in both the current-carrying and the kinetic properties of the channels. Under all our experimental conditions, i.e., in external solutions of normal or low ionic strength, with or without external divalent cations, and at external pHs between 5.5 and 8.1, TMO treatment reduced ACh single-channel conductance to 70-90% of normal. The effects of TMO on channel kinetics were dependent on the ionic conditions. In normal ionic strength solutions containing both calcium and magnesium ions TMO modification reduced the channel average open time by approximately 25%. A similar reduction in open time was seen in calcium-free solution, but was not present when both calcium and magnesium ions were absent from the external solution. Lowering the ionic strength of the solution increased the mean open time in normal channels by about threefold, but did not affect the kinetics of modified channels. In low ionic strength solutions normal ACh channel open times were maximal at approximately pH 6.7 and decreased by three- to fourfold at both acid and alkaline pH. TMO modification removed the pH dependence of channel kinetics, and average open times were short at all pHs between 5.5 and 8.1. We suggest that TMO modifies normally titratable groups on the external surface of ACh channels that help to determine both the gating and permeability properties of ACh channels.

2006 ◽  
Vol 19 (3) ◽  
pp. 683-696 ◽  
Author(s):  
N. Choucair ◽  
V. Laporte ◽  
R. Levy ◽  
C. Tranchant ◽  
J.-P. Gies ◽  
...  

Author(s):  
Л.А. МАРЧЕНКО ◽  
Т.Н. БОКОВИКОВА ◽  
Е.В. ЛИСОВАЯ ◽  
С.А. ИЛЬИНОВА ◽  
Е.П. ВИКТОРОВА

Исследована возможность применения яблочной кислоты (ЯК) в качестве гидратирующего агента для перевода негидратируемых форм фосфолипидов в гидратируемые. Выбор ЯК обусловлен ее стоимостью, которая в 1,5 раза меньше стоимости янтарной кислоты, широко применяемой в качестве гидратирующего агента, а также большей доступностью с точки зрения промышленного производства. Исследование процесса комплексообразования и определение состава комплексных соединений ЯК с ионами кальция и магния осуществляли методом потенциометрического титрования. Установлено, что внесение в раствор ЯК ионов кальция и магния приводит к снижению значений рН, что свидетельствует о наличии комплексообразования в указанных системах. Наиболее устойчивыми являются комплексы ионов кальция и магния с непротонированным лигандом при соотношении Ме2 : лиганд 1 : 1. В процессе комплексообразования ионы Ca2 и Mg2 вытесняют протоны только карбоксильных групп ЯК, которая с ионами кальция образует более устойчивые комплексы, чем с ионами магния. Показано, что устойчивость комплексов ЯК с ионами кальция и магния значительно выше, чем устойчивость комплексов фосфатидилсеринов и фосфатидных кислот с указанными ионами. Использование водных растворов ЯК в качестве гидратирующего агента позволит повысить эффективность процесса гидратации и увеличить выход фосфолипидов и, следовательно, готового продукта лецитина. The possibility of using malic acid (MA) as a hydrating agent for converting non-hydrated forms of phospholipids into hydrated ones has been investigated. The choice of MA is due to its cost, which is 1,5 times less than the cost of succinic acid, as well as greater availability from the point of view of industrial production. The study of the complexation process and determination of the composition of complex compounds of MA with calcium and magnesium ions was carried out by the method of potentiometric titration. It was found that the introduction of calcium and magnesium ions into the MA solution leads to a decrease in pH values, which indicates the presence of complexation in these systems. Complexes of calcium and magnesium ions with an unprotected ligand at a ratio of ME2 : ligand 1 : 1 are the most stable. In the process of complexing, Ca2 and Mg2 ions displace protons only of the carboxyl groups of MA, which forms more stable complexes with calcium ions than with magnesium ions. It is shown that the stability of MA complexes with calcium and magnesium ions is significantly higher than the stability of phosphatidylserine and phosphatidic acid complexes with these ions. Using water solutions of MA as a hydrating agent will increase the efficiency of the hydration process and increase the yield of phospholipids and, consequently, the finished product lecithin.


Author(s):  
Aurica Pop ◽  

The paper showcases research conducted with the purpose of determining the Ca2+ and Mg2+ ions in the drinking water (fountain water) of a common household located in Dumbravita, Maramures County, Romania. The photometric method was used for this goal, as well as a Calcium and Magnesium Photometer. This study argues about implementing cost efficient techniques and precise analysis in order to determine Ca and Mg ions in the water samples.


1974 ◽  
Vol 60 (3) ◽  
pp. 673-705 ◽  
Author(s):  
A. N. CLEMENTS ◽  
TERRY E. MAY

1. Two nerve-muscle preparations were used to investigate the physiology of the locust retractor unguis muscle in relation to L-glutamic acid. These were an ‘isolated preparation’, in which the muscle and its nerve were dissected out, and a ‘perfusedfemur preparation’, in which the muscle suffered no mechanical disturbance. 2. Exposure of the nerve--muscle preparations to glutamate caused a variety of responses, some of which were shown to be abnormal and due to the experimental conditions. 3. When locust femora were perfused with saline or haemolymph the retractor unguis muscles were much more severely affected by glutamate if the hydrostatic pressure was slightly raised. At raised pressures the perfused-femur preparations were particularly prone to give repetitive and spontaneous contractions. 4. Analysis of haemolymph from adult male locusts showed that it contained, on average, 0-2 mmol/1 L-glutamate, 45 mol/1 total non-peptide amino acids, 5-0 mmol/1 calcium, and 11-6 mmol/1 magnesium. It was calculated that approximately 50% of the calcium and 75% of the magnesium ions are bound to amino acids, and that approximately 25% of the glutamic acid is bound to divalent metal ions. 5. The isolated preparations were severely affected by glutamate at the concentration at which it occurs in haemolymph, and it was concluded that in the intact locust some mechanism must protect the neuromuscular synapses from haemolymphg lutamate. No evidence could be obtained of the sequestration of glutamate by haemocytes, or of binding of glutamate to haemolymph proteins. 6. Calcium and magnesium ions reduced the sensitivity of nerve-muscle preparations to glutamate to a greater extent than could be accounted for by the formation of amino acid-metal complexes. This suggests that the protection afforded by calcium and magnesium involves an interaction of the metal ions with the neuromuscular system itself. 7. The retractor unguis muscle was much less sensitive to glutamate when it was contained within an undissected femur than in an isolated preparation. It was concluded that the muscle is normally protected from haemolymph glutamate by a diffusion barrier which is damaged on dissection. 8. Comparison of the fine structure of retractor unguis muscles, fixed either after dissection or while still contained within the femur, showed that dissection normally caused a partial separation of muscle fibres and damage to the connective tissue sheath, with the resultant exposure of some nerve endings. The connective tissue sheath may constitute the postulated diffusion barrier. 9. The excitatory synapses of the locust retractor unguis muscle are believed to be isolated from haemolymph glutamate by a diffusion barrier, which is tentatively identified with the connective tissue sheath that binds the muscle fibres together. Calcium and magnesium ions reduce the sensitivity of nerve-muscle preparations to glutamate, and may have such a role in the living insect.


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