Further observations of aldosterone response in barnacle muscle fibers

1983 ◽  
Vol 245 (3) ◽  
pp. E230-E238
Author(s):  
R. B. Tallitsch

Work with single muscle fibers from the barnacle Balanus nubilus has revealed that these fibers can be rendered sensitive to external application of aldosterone by preexposing the barnacle in vivo to the steroid for 16 h. Experiments investigating the dose-response relationship, glucocorticoid sensitivity of the preparation, and the role of the mitochondria in the aldosterone response in this preparation are presented. It is demonstrated that saturation kinetics may be seen in the dose-response curve if care is taken to isolate only fibers from animals that are in midmolt cycle. Data is presented that demonstrated a specific mineralocorticoid response to aldosterone in this preparation rather than a combined mineralocorticoid-glucocorticoid response, as has been demonstrated in other preparations. Experiments investigating the role of the mitochondrial inhibitors and substrates suggest that aldosterone modulates Na efflux in barnacle muscle fibers by altering mitochondrial function, presumably at the level of succinic hydrogenase.

1975 ◽  
Vol 53 (2) ◽  
pp. 285-292 ◽  
Author(s):  
Stephen S. Chen

Benzocaine, which occurs in the uncharged form in the physiological range of pH, caused inhibition of 45Ca efflux in barnacle muscle fibers. By contrast, in the presence of a low external Ca2+ concentration, it produced stimulation of the efflux. Both the inhibitory and stimulatory actions of benzocaine appeared to be less potent than those of procaine. Hemicholinium-3 (HC-3), on the other hand, which exists only in the charged form, caused a large stimulation of the 45Ca efflux following microinjection, and the potency of this action was found to be at least 10 times greater than that of procaine. External application of HC-3 produced inhibition occasionally. Effects of tetracaine were similar to those produced by procaine; however, its inhibitory action was greater in more alkaline solution, which is the opposite of that observed with procaine. Lidocaine produced a less consistent effect than procaine; the inhibitory action of the former was less potent but the stimulatory action of the two anesthetics were comparable. p-Aminobenzoic acid was without effect on 45Ca efflux. These results indicate that both the charged and uncharged forms of local anesthetics are capable of causing stimulatory and inhibitory effects on 45Ca efflux in barnacle muscle fibers, and that the inhibition produced is the result of action on the Ca–Ca exchange system whereas the stimulation is the result of release of Ca from internal storage sites.


1981 ◽  
Vol 240 (1) ◽  
pp. C80-C89 ◽  
Author(s):  
W. F. Boron ◽  
W. C. McCormick ◽  
A. Roos

Intracellular pH (pHi) regulation was studied in barnacle muscle fibers with pH-sensitive microelectrodes. The cells were acid loaded, and the subsequent recovery of pHi was monitored. The rate of recovery was reduced by one-third when external Na+ ([Na+]o) was replaced by Li+, but recovery was completely abolished when Na+ was replaced by choline or N-methyl-D-glucamine. In other experiments, varying amounts of Na+ were replaced by choline, and the acid extrusion rate, derived from the recovery rate of pHi, was calculated at a single value of pHi, 6.80. The dependence of the acid extrusion rate on [Na+]o could be described by Michaelis-Menten kinetics; at pHo (extracellular) = 8.0 and [HCO3-]o (extracellular) = 10 mM, the apparent Km and Vmax were 59 mM and 1.3 mmol x l(-1) x min-1. When [HCO3-]o was reduced to 2.5 mM at the same pHo, Km did not change significantly, but Vmax was substantially reduced. On the other hand, when pHo was reduced to 7.4 at constant [HCO3-]o, Vmax changed only slightly, but Km increased substantially. In similar experiments, we examined the dependence of the acid extrusion rate on [HCO3-]o. At pHo = 8.0 and [Na+]o = 440 mM, the apparent Km and Vmax were 4.1 mM and 2.1 mmol x 1-1 x min-1. When pHo was reduced to 7.4, Vmax was not altered, but Km substantially increased. The kinetic data are discussed in terms of the role of pHo, [Na+]o, and [HCO3-]o in the pHi-regulating system.


1992 ◽  
Vol 263 (1) ◽  
pp. C246-C256 ◽  
Author(s):  
B. A. Davis ◽  
E. M. Hogan ◽  
W. F. Boron

We internally dialyzed single barnacle muscle fibers (BMF) for 90 min with a dialysis fluid (DF) containing no Na+ and either 0 or 100 mM Li+ and measured intracellular pH (pHi) with a microelectrode. During dialysis, the pH 8.0 artificial seawater (ASW) contained neither Na+ nor HCO3-. After we halted dialysis with a Li(+)-free/low-pH DF and allowed pHi to stabilize at approximately 6.8, adding 440 mM Na(+)-10 mM HCO3- to the ASW caused pHi to recover rapidly and stabilize at 7.32. In contrast, when the DF contained 100 mM Li+, pHi stabilized at 7.49. In fibers dialyzed to a pHi of approximately 7.2, Li+ stimulated a component of acid extrusion that was dependent on Na+ but not affected by SITS. Thus Li+ activates a Na(+)-dependent acid-extrusion mechanism other than the well characterized Na(+)-dependent Cl-HCO3 exchanger. To study the Li(+)-activated mechanism, we minimized Na(+)-dependent Cl-HCO3 exchange by raising pHDF to 7.35 and pretreated BMFs with SITS. We found that dialysis with Li+ elicits a Na(+)-dependent pHi increase that is largely blocked by amiloride, consistent with the hypothesis that Li+ activates a latent Na-H exchanger even at a normal pHi. In the absence of Li+, the Na-H exchanger is relatively inactive at pHi 7.35 (net acid-extrusion rate, Jnet = 9.5 microM/min) but modestly stimulated by reducing pHi to 6.8 (Jnet = 64 microM/min). In the presence of Li+, the Na-H exchanger is very active at pHi values of both 7.35 (Jnet = 141 microM/min) and 6.8 (Jnet = 168 microM/min). Thus Li+ alters the pHi sensitivity of the Na-H exchanger. Because the Na-H exchanger is only approximately 6% as active as the Na(+)-dependent Cl-HCO3 exchanger in the absence of Li+ at a pHi of approximately 6.8, we suggest that the major role of the Na-H exchanger may not be in pHi regulation but in another function such as cell-volume regulation.


1976 ◽  
Vol 30 (1) ◽  
pp. 197-212
Author(s):  
Daniel F. Wolff ◽  
Osvaldo A. Alvarez ◽  
Fernando F. Vargas

1983 ◽  
Vol 244 (3) ◽  
pp. C297-C302 ◽  
Author(s):  
S. S. Sheu ◽  
M. P. Blaustein

The influence of internal and external Ca2+ on membrane potential and 22Na influx were tested in internally perfused giant barnacle muscle fibers. The fibers depolarized by about 2-3 mV, and Na+ influx increased when external Ca2+ was removed. These effects were inhibited and reversed by adding 2 mM La3+ externally but not by tetrodotoxin (TTX). Ca2+ channel blockers did not prevent the depolarization. Increasing internal free Ca2+ ([Ca2+]i) from 10(-7) to 10(-5) M also stimulated Na+ influx and depolarized the fibers by a few millivolts. Neither external La3+ nor TTX prevented the effects of raising [Ca2+]i; however, internal tetrabutylammonium ions depolarized the fibers and attenuated the internal Ca2+-dependent effects. These data are consistent with the idea that removal of external Ca2+ activates a La3+-sensitive channel that is permeable to Na+; raising [Ca2+]i activates a La2+-insensitive, Na+-permeable channel that may be similar to the internal Ca2+-activated nonselective cation channels observed in cardiac muscle. The results demonstrate that all Na+ (and Ca2+) fluxes that do not contribute to Na-Ca exchange must be carefully identified before the exchange stoichiometry can be determined from Na+ and Ca2+ flux measurements.


1990 ◽  
Vol 78 (1) ◽  
pp. 101-104 ◽  
Author(s):  
Joe Collier ◽  
Patrick Vallance

1. The dose-response to acetylcholine has been examined in dorsal hand veins of healthy volunteers before and after removal of the endothelium. 2. Measurements were made in single dorsal hand veins during local infusions of acetylcholine. The vein was irrigated with distilled water to remove the endothelium. Dilator studies were performed in vessels preconstricted by a continuous infusion of noradrenaline. 3. In the endothelium-intact vessel the dose-response to acetylcholine was biphasic; low doses produced venodilatation with higher doses causing venoconstriction. 4. Dilatation to low doses of acetylcholine was abolished by prior irrigation with distilled water, consistent with denudation of the endothelium by this process. Irrigation augmented the constriction seen in response to higher doses of acetylcholine. 5. This is the first demonstration of an endothelium-dependent biphasic dose-response to acetylcholine in man. The results raise questions as to the possible physiological actions of endogenous acetylcholine and as to the use of the acetylcholine dose-response curve as a marker of endothelial function.


1987 ◽  
Vol 65 (7) ◽  
pp. 1416-1420 ◽  
Author(s):  
Jean-Pierre Caillé ◽  
Marie Pigeon-Gosselin ◽  
Michel Pézolet

The Raman spectra observed from barnacle muscle fibers are quite complex because the cytoplasm of these cells contains several proteins and solutes. An extraction procedure was used to separate organic solutes from the contractile proteins. Glycine, trimethylamine oxide, taurine, and alanine were found to contribute to the Raman spectra of barnacle muscle fibers, while spectra of lobster fibers reveal the presence of betaine in addition. We have observed that the increase in osmolarity of the intracellular fluid caused by the augmentation of the salinity of sea water (density, 1.023–1.030) in which the barnacles were kept, induces a reduction of intensity of the amide I band. To distinguish among the different parameters which are modified by the sea water salinity, observations were made on glycerinated barnacle muscle fibers. The reduction of intensity of the amide I band in the Raman spectra of glycerinated muscle fibers was also observed with the addition of taurine (0.08 M) in the external relaxing solution. Therefore, under these experimental conditions, the Raman scattering intensity in the amide I region assigned to the α-helix conformation (1645–1650 cm−1) is increased when the concentration of organic electrolytes is reduced. However, as no significant decrease of the scattering intensity in the 1660–1670 cm−1 region where the amide I bands of either β-sheet or disordered conformations normally appear was observed, the increase of intensity of the amide I band centered at 1645 cm−1 is assigned to a change of orientation of α-helical segments of the myosin molecules. Our results suggest that organic solutes influence the position of the S-2 segments relative to the thick filaments.


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