ChemInform Abstract: SYNTHESES OF FLUORESCENT DYES. XIV. STANDARDS FOR FLUORESCENCE MEASUREMENTS IN THE NEAR NEUTRAL PH‐RANGE. (1)

Wolfbeis, Otto S.; Urbano, Edmund

doi:10.1002/chin.198311287

Cited by 6 publications

(9 citation statements)

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“…Storage at -80~ had no effect on transport characteristics. SPQ was synthesized as described previously [30], Vesicles ( -8 mg protein/ml) were loaded with 10 mM SPQ and appropriate buffers by incubation for > 24 hr at 4~ Buffers contained 100 mM sucrose, 10 mM HEPES-Tris, X mM K-gluconate, and (150 -X) mM N-metbyl-D-glucamine gluconate (NMG-glucohate), pH 7.0. X = 50 mM in experiments where membrane potential (4') was damped at 0 mV; X = 5 mM when ~ was set to +60 inV.…”

Section: Methodsmentioning

confidence: 99%

Chloride transport in apical membrane vesicles from bovine tracheal epithelium: Characterization using a fluorescent indicator

1988

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C1 transport in apical membrane vesicles derived from bovine tracheal epithelial cells was studied using the C1-sensitive fluorescent indicator 6-methoxy-N-(3-sulfopropyl) quinolinium. With an inwardly directed 50 mM C1 gradient at 23 degrees C, the initial rate of C1 entry (JC1) was increased significantly from 0.32 +/- 0.12 nmol.sec-1.mg protein-1 (mean +/- SEM) to 0.50 +/- 0.07 nmol.sec-1.mg protein-1 when membrane potential was changed from 0 to +60 mV with K/valinomycin. At 37 degrees C, with membrane potential clamped at 0 mV, there was a 34 +/- 7% (n = 5) decrease in JC1 from a control value of 0.37 +/- 0.03 nmol.sec-1.mg protein-1 upon addition of 0.2 mM diphenylamine-2-carboxylate. The following did not alter JC1 significantly (JC1 values given as percent change from control): 50 mM cis Na (-1 +/- 5%), 0.1 mM furosemide (-3 +/- 4%), 0.1 mM furosemide in the presence of 50 mM cis Na (-5 +/- 2%), 0.1 mM H2DIDS (-18 +/- 9%), a 1.5 pH unit inwardly directed H gradient (-7 +/- 7%), and 0.1 mM H2DIDS in the presence of a 1.5 unit pH gradient (4 +/- 18%). With inward 50 mM anion gradients, the initial rates of Br and I entry (JBr and JI, respectively) were not significantly different from JC1.JC1 was a saturable function of C1 concentration with apparent Kd of 24 mM and apparent Vmax of 0.54 nmol.sec-1.mg protein-1.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Methodsmentioning

confidence: 99%

Chloride transport in apical membrane vesicles from bovine tracheal epithelium: Characterization using a fluorescent indicator

1988

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“…SPQ, synthesized by the method of Wolfbeiss and Urbano [11], was the generous gift of Jacqui Marshall, Department of Biology, University of York. SPQ was dissolved directly into culture medium to a strength of 20 mM prior to each experiment.…”

Section: Chemicalsmentioning

confidence: 99%

“…Thus SPQ exhibits good C1-selectivity, and under physiological conditions emits fluorescence in relation to a particular free C1-concentration. SPQ was initially synthesized by Wolfbeis and Urbano [11], and first used to measure biological C1-fluxes by Illsley and Verkman [10], in sealed human erythrocyte ghosts and placental microvillous vesicles. It has since been used in a range of biological systems, from plant membrane vesicles [12] to cultured tracheal cells [13,14] and whole isolated rabbit proximal convoluted tubules [15].…”

Section: Introductionmentioning

confidence: 99%

Role of Chloride Transport in the Development of the Rat Blastocyst1

Brison

Leese

1993

Biology of Reproduction

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Preimplantation mammalian development culminates in the generation of a fluid-filled cavity, the blastocoel, which requires the vectorial transport of ions across the trophectoderm, followed by the movement of water. Experiments were carried out to establish the role of Cl- transport in blastocoel formation in the rat. These included investigations of the effect of Cl- substitution and the Cl- transport inhibitors 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, furosemide, and anthracene-9-carboxylic acid, on the development of morulae into blastocysts in culture, and on the rate at which blastocoel fluid is accumulated. In addition, a novel technique was developed in which the Cl(-)-sensitive fluorescent dye 6-methoxy-N-(3-sulfopropyl) quinolium (SPQ) was loaded into the blastocoel cavity, in order to characterize the pathways of transtrophectodermal Cl- transport. We established that 1) blastocoel formation in the rat requires the external presence of Cl- ions; 2) transport across the trophectoderm appears to be via a transcellular pathway, since rates of blastocyst development, fluid accumulation, and SPQ-monitored Cl- efflux are all reduced in the presence of Cl- transport inhibitors; and 3) Cl- transport occurs via Cl-/HCO3- exchange.

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“…Storage roots were harvested immediately before use. The fluorescent probe, SPQ, was synthesised and purified as described previously (Wolfbeis and Urbano 1982;Pope and Leigh 1988). Amiodarone was a gift from Sanofi UK (Wythenshawe, Manchester, UK).…”

Section: Methodsmentioning

confidence: 99%

Characterisation of chloride transport at the tonoplast of higher plants using a chloride-sensitive fluorescent probe

Pope¹,

Leigh²

1990

Planta

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The characteristics of Cl(-) transport in isolated tonoplast vesicles from red-beet (Beta vulgaris L.) storage tissue have been investigated using the Cl(-)-sensitive fluorescent probe, 6-methoxy-1-(3-sulfonatopropyl)-quinolinium (SPQ). The imposition of (inside) positive diffusion potentials, generated with K(+) and valinomycin, increased the initial rate of Cl(-) transport, demonstrating that Cl(-) could be electrically driven into the vesicles. Chloride influx was unaffected by SO 4 (2-) , but was competitively blocked by NO 3 (-) , indicating that both Cl(-) and NO 3 (-) may be transported by the same porter. In some preparations, increases in free-Ca(2+) concentration from 10(-8) to 10(-5) mol·dm(-3) caused a significant decrease in Cl(-) influx, which may indicate that cytosolic Ca(2+) concentration has a role in controlling Cl(-) fluxes at the tonoplast. However, this effect was only seen in about 50% of membrane preparations and some doubt remains over its physiological significance. A range of compounds known to block anion transport in other systems was tested, and some partially blocked Cl(-) transport. However, many of these inhibitors interfered with SPQ fluorescence and so only irreversible effects could be tested. The results are discussed in the context of recent advances made using the patch-clamp technique on isolated vacuoles.

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ChemInform Abstract: SYNTHESES OF FLUORESCENT DYES. XIV. STANDARDS FOR FLUORESCENCE MEASUREMENTS IN THE NEAR NEUTRAL PH‐RANGE. (1)

Abstract: Die mit dem Sulton (V) aus entsprechenden Ausgangsmaterien erhaltenen Farbsalze (I)‐(IV) werden hinsichtlich ihrer fluoreszierenden Wirkung quantitativ untersucht.

Cited by 6 publications

References 1 publication

Chloride transport in apical membrane vesicles from bovine tracheal epithelium: Characterization using a fluorescent indicator

Chloride transport in apical membrane vesicles from bovine tracheal epithelium: Characterization using a fluorescent indicator

Role of Chloride Transport in the Development of the Rat Blastocyst1

Characterisation of chloride transport at the tonoplast of higher plants using a chloride-sensitive fluorescent probe

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