Ratio confocal imaging of free cytoplasmic calcium gradients in polarising and polarisedFucuszygotes

Zygote ◽  
1993 ◽  
Vol 1 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Frederic Berger ◽  
Colin Brownlee

SummaryIn the marine brown alga,Fucus, two poles are differentiated before cell division determining the future rhizoid or thallus. We have used a combination of the Ca2+-sensitive dye Calcium Green and the pH-sensitive dye SNARF monitored at pH-insensitive wavelengths to obtain confocal ratio images of free cytoplasmic calcium distribution at different stages in polarisingFucuszygotes. These dyes have the advantage that they can be used in most confocal microscopes and their longer excitation wavelengths greatly reduce autofluorescence problems. Dyes of varying molecular weights (free acid form, 10 000 mol.wt or 70 000 mol.wt dextran-conjugated) were pressure microinjected into early zygotes which were allowed to polarise in unidirectional light. Dextran-conjugated dyes remained non-compartmentalised and fluorescence could be monitored for up to 3 days following microinjection. Currently we have been able to detect Ca2+gradients at the tip of the rhizoid, confirming earlier results. Localised Ca2+elevations have also been observed at the rhizoid pole of the polarising zygote before the onset of rhizoid germination. Limitations of this technique and the significance of these Ca2+gradients are discussed.

1989 ◽  
Vol 257 (2) ◽  
pp. C297-C305 ◽  
Author(s):  
E. Honore ◽  
C. Martin ◽  
C. Mironneau ◽  
J. Mironneau

The whole cell voltage-clamp technique was used to study the effects of extracellular ATP in cultured smooth muscle cells isolated from pregnant rat myometrium. An inward current was elicited by ATP (IATP) in cells held at -70 mV under voltage clamp. The amplitude of IATP was reduced by estrogen pretreatment and by the end of pregnancy. IATP not only did not undergo any desensitization but showed facilitation. The current-voltage relationship of IATP was linear and reversed close to 0 mV. Changing the sodium electrochemical gradient by decreasing extracellular or intracellular sodium resulted in a linear relationship between the reversal potential of IATP and Na equilibrium potential that, however, differed from the predicted curve for a purely sodium conductance. The conductance activated by ATP was monovalent cation selective with little discrimination between potassium, cesium, and sodium ions. IATP was depressed by divalent cations, and the rank order of potency was Co greater than Mg greater than Ca greater than Ba, suggesting that the free-acid form of ATP was the effective ligand. Adenosine, AMP, and ADP were ineffective in eliciting IATP, whereas ATP gamma S and alpha,beta-methylene ATP were capable of mimicking the effects of ATP, although they were less potent. These results are consistent with the free-acid form of ATP activating a monovalent cation-selective and estrogen-sensitive conductance in myometrium.


ChemInform ◽  
2008 ◽  
Vol 39 (7) ◽  
Author(s):  
Hideyuki Murakami ◽  
Kunihiko Hayashi ◽  
Ikuo Tsukada ◽  
Takeshi Hasegawa ◽  
Shoko Yoshida ◽  
...  

The Analyst ◽  
2020 ◽  
Vol 145 (11) ◽  
pp. 4004-4011
Author(s):  
Minghui Yi ◽  
Linchuan Zhao ◽  
Kang Wu ◽  
Chang Liu ◽  
Diandian Deng ◽  
...  

Methyl jasmonate (MeJA) and its free-acid form, jasmonic acid (JA), collectively referred to as jasmonates (JAs), are natural plant growth regulators that are widely present in higher plants.


1994 ◽  
Vol 47 (4) ◽  
pp. 677
Author(s):  
GL Mendz

Fluorinated derivatives of fluorescent calcium probes have been recently used to measure free calcium concentrations in biological fluids by 19F n.m.r. spectroscopy. To measure calcium levels a method utilizes the change in chemical shift experienced by the fluorine resonance of the calcium chelator 1,2-bis(2-amino-5-fluorophenoxy)ethane-N,N,N′,N′- tetraacetic acid upon complexation of cations. The procedure assumes that (A) the fluorinated probe is in equilibrium between the calcium complex and the free acid form; (B) the chemical exchange between different sites has similar effects on the intensities of the observable resonances; and (c) the presence of macromolecules affects equally the intensities of the resonances arising from different complexes. Experiments designed to test these assumptions were carried out in plasma and in model systems containing mixtures of cations and macromolecules. The results indicated that in biological fluids the calcium probe is involved in a multisite equilibrium in which 'invisible' (very broad) peaks affect the equilibrium intensities of the 'observable' peaks. Binding to macromolecules affected differently the intensities of the resonances arising from different complexes in the spectrum of the chelator , and influenced significantly the calculated free calcium concentrations.


2007 ◽  
Vol 80 (11) ◽  
pp. 2161-2169 ◽  
Author(s):  
Hideyuki Murakami ◽  
Kunihiko Hayashi ◽  
Ikuo Tsukada ◽  
Takeshi Hasegawa ◽  
Shoko Yoshida ◽  
...  

1981 ◽  
Vol 61 (1) ◽  
pp. 111-113 ◽  
Author(s):  
B. A. Scoggins ◽  
J. G. McDougall ◽  
Aldona Butkus ◽  
J. P. Coghlan ◽  
D. A. Denton ◽  
...  

1. To test the hypothesis that [Asp1,Val5]-angiotensin-(1–8)octapeptide ([Asp1,Val5]ANG II) is a more potent agonist to aldosterone secretion in the sodium-depleted animal than is [Asn1,Val5]angiotensin-(1–8)octapeptide ([Asn1,Val5]ANG II), local adrenal arterial infusion of the two peptides has been carried out in sheep with cervical adrenal autotransplants. 2. Neither [Asp1,Val5]ANG II nor [Asn1,Vak5]-ANG II further stimulated the increased level of aldosterone secretion in conscious moderately sodium-depleted sheep. Greater sodium deficiency further increased aldosterone secretion. 3. The conclusion of Campbell, Schmitz & Itskovitz [Clinical Science (1979), 56, 325–333] that the free acid form of angiotensin II was a more potent agonist of aldosterone secretion than was the amide form under conditions of reduced sodium status is not supported by studies in sodium-depleted sheep.


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