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W J Gibson

Publications and source records attributed to W J Gibson.

15 recordsLinked to original sources
PubMed2000

The interaction of U-73122 with the histamine H1 receptor: implications for the use of U-73122 in defining H1 receptor-coupled signalling pathways.

U-73122, an N-aminosteroid homologue of N-ethylmaleimide (NEM), widely used as an inhibitor of phospholipase C, was found to be a potent inhibitor (IC50 5.5+/-0.5 microM) of the binding of [3H]mepyramine to guinea-pig cerebellar membranes. The succinimido analogue, U-73343, also inhibited the binding of [3H]mepyramine (estimated IC50 24+/-1 microM), but NEM was only a weak inhibitor, even at 10 mM. The interaction of U-73122 and U-73343 with the H1 receptor was effectively irreversible, on the time-scale of the experiment. There is no indication that reaction with a receptor thiol residue is involved in the binding of U-73122, since preincubation of membranes with 2 mM NEM did not significantly increase the IC50 for the inhibition of [3H]mepyramine binding by U-73122. We conclude that U-73122 binds to the histamine H1 receptor in the concentration range in which it acts as an inhibitor or PLC. This compromises the use of U-73122 to provide evidence that an H1 agonist action is mediated via PLC. The tight binding of U-73343 to the receptor appears to be primarily a function of the hydrophobic nature of the compound.

Animals↗
PubMed1998

SK&F 96365 (1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenylethyl]-1H- imidazole hydrochloride) stimulates phosphoinositide hydrolysis in human U373 MG astrocytoma cells.

SK&F 96365 (1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenylethyl]-1H-imi dazole hydrochloride) stimulated the accumulation of [3H]inositol monophosphates ([3H]IP1) in human U373 MG astrocytoma cells prelabelled with [3H]inositol (EC50 15 +/- 1 microM, Hill coefficient 3.8 +/- 0.4). SK&F 96365-induced accumulation of [3H]IP1 increased linearly with time, but there was no initial rapid formation of [3H]IP3. SK&F 96365 also stimulated [3H]IP1 accumulation in human HeLa cells, but only to a small extent in slices of rat cerebral cortex and guinea-pig cerebellum. SK&F 96365-induced accumulation of [3H]IP1 in U373 MG cells increased as extracellular Ca2+ was increased from nominally zero to 4 mM, but there was no evidence that SK&F 96365 induced any marked entry of Ca2+ into cells; only an inhibition of store-refilling-induced Ca2+ entry was apparent. Further, the response to SK&F 96365 was additive with that to the Ca2+ ionophore ionomycin. Depolarization of the cells with raised K+ produced only a small stimulation of phosphoinositide hydrolysis. SK&F 96365 caused the release of Ca2+ from intracellular stores in U373 MG cells (EC50 26 +/- 14 microM), but thapsigargin induced only a small accumulation of [3H]IP1. Miconazole, another N-substituted imidazole, also stimulated [3H]IP1 accumulation in U373 cells.

Animals↗
PubMed1998

Dual effects of histamine and substance P on intracellular calcium levels in human U373 MG astrocytoma cells: role of protein kinase C.

1. In human U373 MG astrocytoma cells agonist-induced increases in intracellular Ca2+ ([Ca2+]i) are rapidly returned towards prestimulated levels. Examination of the effect of histamine and substance P on [Ca2+]i in thapsigargin-treated cells has allowed a mechanism contributing to this effect to be characterized. 2. Histamine and substance P stimulated [3H]-inositol monophosphate ([3H]-IP1) accumulation in U373 MG cells. Concentration-response curves of [3H]-IP1 accumulation in suspensions of U373 MG cells in HEPES buffer containing 30 mM Li+ yielded best-fit EC50 values of 19.1+/-1.5 microM for histamine and 5.7+/-1.3 nM for substance P. 3. In confluent monolayers of fura-2 loaded U373 MG cells perfusion with 100 microM histamine resulted in a transient 597+/-50 nM increase in [Ca2+]i. The best-fit EC50 for histamine was 4.6+/-2.2 microM. The initial, transient, histamine response was often followed by further small transient increases in [Ca2+]i. 4. Treatment of U373 MG cells with 5 microM thapsigargin, followed by the readdition of 1.8 mM Ca2+ to the perfusion buffer, resulted in a steady-state level of [Ca2+]i 97+/-5 nM above pretreated levels (measured 400 s after readdition of Ca2+). Perfusion of histamine (100 microM, 100 s) caused a rapid decline in the thapsigargin-induced steady state level of [Ca2+]i. This effect of histamine was normally reversible upon washout. The best-fit EC50, for the histamine response was 0.8+/-0.2 microM. Substance P (10 nM, 100s) also caused a reduction in thapsigargin-induced steady-state levels of [Ca2+]i. 5. Neither 100 microM histamine nor 10 nM substance P inhibited the rate of quench of fura-2 fluorescence by Mn2+ in U373 MG cells pretreated with 5 microM thapsigargin, indicating that the depressant effect on steady-state raised [Ca2+]i was probably not due to a block of Ca2+ entry. 6. The depressant effect of histamine on [Ca2+]i was blocked by 1 microM mepyramine, and was partially reduced by pre-incubation with 1 microM staurosporine (61+/-7% reduction) and with Ro 31-8220 (24+/-10% and 50+/-6% reduction by 1 and 10 microM Ro 31-8220, respectively). Pre-incubation with H-89 did not alter the depressant effect of histamine. 7. Neither 1 microM staurosporine nor 10 microM KN-62 inhibited the binding of [3H]-mepyramine to guinea-pig cerebellar membranes, whereas it was reduced by 17+/-1% and 55+/-2% by 1 and 10 microM Ro 31-8220, respectively. However, [3H]-IP1 accumulation stimulated by histamine in U373 MG cells was not inhibited by 1 or 10 microM Ro 31-8220 and in 2 out of 3 experiments there was a significant potentiation of the response to histamine with both concentrations of Ro 31-8220. Staurosporine, 1 microM, similarly potentiated the response to 100 microM histamine in 3 out of 4 experiments. KN-62 (10 microM) did not stimulate histamine-induced [3H]-IP1 accumulation. 8. In HEPES buffer to which no Ca2+ had been added, histamine stimulated a transient 451+/-107 nM increase in [Ca2+]i. Pretreatment with 1 microM and 10 microM Ro 31-8220 did not significantly alter the initial peak response to histamine, but slowed the rate at which histamine-induced increases in [Ca2+]i were returned to prestimulated levels. Pretreatment with KN-62 had no significant effect on the response to histamine, but consistently inhibited the secondary slower phase of the decline in [Ca2+]i. H-89 did not alter the histamine response. 9. The effect of histamine in stimulating Ca2+ extrusion was not confined to U373 MG cells, since 100 microM histamine also caused a rapid decrease in steady-state levels of [Ca2+]i in thapsigargin-treated human HeLa cells. 10. The results indicate that agonists which increase [Ca2+]i via activation of phosphoinositide metabolism can also stimulate a homeostatic mechanism which acts to reduce [Ca2+]i. The balance of the evidence indicates that in U373 MG cells the latter effect most likely involves a PKC-mediated stimulation of a Ca2+-extrusion pump.

Animals↗
PubMed1994

Modulation of antagonist binding to histamine H1-receptors by sodium ions and by 2-amino-2-hydroxymethyl-propan-1,3-diol HCl.

1. NaCl (100 mM) reduced the potency of (+)-N-methyl-4-methyldiphenhydramine ((+)-QMDP) as an inhibitor of the binding of [3H]-mepyramine to histamine H1-receptors on guinea-pig cerebellar membranes to a greater extent than that of mepyramine, consistent with the greater inhibitory effect of Na+ on the binding of [3H]-QMDP than on the binding of [3H]-mepyramine. 2. The concentration of 2-amino-2-hydroxymethyl-propan-1,3-diol HCl (Tris, HCl) buffer, pH 7.5, present had little effect on the temelastine-insensitive binding of [3H]-mepyramine, but caused a concentration-dependent inhibition of the binding of [3H]-mepyramine sensitive to 1 microM temelastine (H1-receptor binding), with an approximate IC50 of 75 mM, assuming that complete inhibition would have been achieved. 3. Inhibition of [3H]-mepyramine binding by Na+ was more marked in 10 mM than in 50 mM Tris HCl and was not evident in 200 mM Tris HCl. 4. The Kd for the temelastine-sensitive binding of [3H]-mepyramine measured in 10 mM Tris HCl, 0.24 +/- 0.01 nM, was increased by 2.2 +/- 0.2 fold by 100 mM NaCl, without any significant change in the maximum binding (Bmax). The Bmax for [3H]-mepyramine was similarly unchanged in 50 mM Tris HCl, but the Kd was increased 2.5 +/- 0.2 fold. 5. The Kd for the temelastine-sensitive binding of [3H]-mepyramine was also increased in 50 mM,compared with 10 mM, N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulphonic acid] KOH (HEPES.KOH)buffer (Kd 0.25 +/- 0.02 nm in 10 mM HEPES), but the evidence for an interaction between HEPES and Na+ was less clear.6. The effect of 100 mM NaCl on the inhibition of [3H]-mepyramine binding in 10 mM Tris HCl was examined for a range of antagonists. The decrease in potency caused by Na+ was greatest for triprolidine, (+)-chlorpheniramine and benzilylcholine (9.6-10.3 fold increase in K1 values) but the binding of mepyramine and promethazine was much less affected (1.8 and 1.9 fold increase in Kd respectively). The Kd for temelastine was not significantly changed. In contrast to the general decrease in antagonist affinity in the presence of Na+, the for MDL 16,455A (4-[1-hydroxy-4-[4-(hydroxydiphenylmethyl)-1-piperidinyl]butyl]-alpha,alpha-dimethylbenzene acetic acid) was increased, but only by 1.5 fold.7. It is concluded that Na+ can act as an allosteric effector of the binding of antagonists at the histamine HI-receptor. Tris HCl also appears to have an allosteric action at the H1-receptor.

Animals↗
PubMed1975

Experimental evaluation of coronary infusates in dogs.

In a study involving dogs, a coronary infusate containing a variety of substances including cardiac substrates, metabolic inhibitors, membrane stabilizers, electrolytes, and anticoagulants was evaluated in terms of protection afforded to the myocardium for up to 90 minutes of normothermic anoxic arrest. Electron microscopical studies and tissue adenosine triphosphate measurements revealed this coronary infusate to be considerably beneficial to the myocardium. Infused myocardium was better preserved compared to myocardium that was not infused or infused with two other "control" infusates. The individual components of our infusate are known cardiac preservatives. The rationale for combining a number of them in our infusate is to produce an additive or synergetic effect through beneficial intervention at several points in the cardiac metabolic cycle. This objective appears to have been fulfilled under the conditions of the present experiment.

Adenosine Triphosphate↗