Mechanisms of pHi control and relationships between tension and pHi in human subcutaneous small arteries

1995 ◽  
Vol 268 (3) ◽  
pp. C580-C589 ◽  
Author(s):  
P. Carr ◽  
W. McKinnon ◽  
L. Poston
Keyword(s):  
Isometric Tension ◽  
Disulfonic Acid ◽  
Physiological Salt Solution ◽  
Ang Ii ◽  
Tension Development ◽  
Small Arteries ◽  
Transport Inhibitor ◽  
Acid Load ◽  
Ethanesulfonic Acid

Intracellular pH (pHi) control and relationships between pHi and tension have been investigated in human subcutaneous small arteries. Isometric tension and pHi (using 2',7'-bis(carboxyethyl)- 5(6)-carboxyfluorescein) were estimated simultaneously. pHi recovery from an acute acid load was dependent on external Na+ and partially inhibited by the absence of HCO3(-) [N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered solution] or by the anion transport inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). In an HCO3(-)-buffered physiological salt solution (PSS), pHi recovery was partially blocked by hexamethylene amiloride (HMA), an inhibitor of Na+/H+ exchange, and completely blocked by DIDS and HMA together. Intracellular Cl- depletion of arteries did not affect the rate of pHi recovery in PSS from an acid load. pHi recovery from acute alkalosis was unaffected by external Na+ removal, reduced in HEPES buffer, and abolished by removal of external Cl-. These data suggest that human small arteries maintain pHi by Na+/H+ exchange and Na(+)-dependent HCO3(-) exchange in response to an acid load, and Na(+)-independent Cl-/HCO3(-) exchange to counteract intracellular alkalosis. Norepinephrine (NE)-, endothelin-1 (ET-1)-, arginine vasopressin (AVP)-, and K(+)-induced tension did not alter pHi in PSS, but there was a small fall with angiotensin II (ANG II). In HEPES, stimulation with K+, NE, ANG II, or AVP led to a fall in pHi, but this did not occur with ET-1. It is therefore unlikely in vivo that an increase in pHi in these arteries would be involved in either tension development or growth induced by these agonists.

Myocardial mechanics predict hemodynamic performance during normal function and alcohol-induced dysfunction in rats

1991 ◽  
Vol 261 (6) ◽  
pp. H1880-H1888
Author(s):  
J. M. Capasso ◽  
P. Li ◽  
P. Anversa
Keyword(s):  
Pressure Rise ◽  
Normal Function ◽  
Left Ventricular ◽  
Isometric Tension ◽  
Papillary Muscles ◽  
Least Squares Regression ◽  
Tension Development ◽  
Myocardial Mechanics

To determine whether mechanical evaluation of muscle tissue removed from the myocardium can be employed as a direct indicator of cardiac contractile performance in situ, isometric and isotonic parameters of muscle mechanics in vitro were correlated with in vivo global functional characteristics of the same heart. Twelve-month-old animals maintained on standard food and water were employed as representative of normal cardiac function. Animals of identical age with left ventricular (LV) dysfunction induced by oral alcohol (30%) ingestion from 4 to 12 mo were utilized to represent depressed cardiac performance. Accordingly, 24 h after the establishment of the hemodynamic profile for a control or experimental heart, the LV posterior papillary muscle was removed from the same heart and examined isometrically and isotonically. Least squares regression analysis was employed to establish a correlation coefficient and P values between various in vitro and in vivo parameters. Hemodynamic measurements were performed under chloral hydrate anesthesia and LV pump performance was evaluated with respect to aortic and ventricular pressures and the rates of rise and decay of the LV pressure trace. Papillary muscles were evaluated with respect to timing parameters of the isometric and isotonic twitch, the first derivative of isometric tension development, and the speed of muscle shortening at increasing physiologic loads. LV peak rate of pressure rise and decay were then correlated with the various isometric and isotonic properties. Myocardial mechanics and hemodynamics revealed depressed function in the papillary muscles and hearts from alcoholic rats. Moreover, significant correlations were found between the LV rate of pressure change (peak +dP/dt and -dP/dt) and both isometric and isotonic twitch measurements.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Timing and magnitude of systolic stretch affect myofilament activation and mechanical work

2014 ◽  
Vol 307 (3) ◽  
pp. H353-H360 ◽  
Author(s):  
Jared R. Tangney ◽  
Stuart G. Campbell ◽  
Andrew D. McCulloch ◽  
Jeffrey H. Omens
Keyword(s):  
Force Transducer ◽  
Isometric Tension ◽  
Mechanical Work ◽  
In Vivo Studies ◽  
Time Varying ◽  
Force Output ◽  
External Work ◽  
Tension Development ◽  
Peak Tension

Dyssynchronous activation of the heart leads to abnormal regional systolic stretch. In vivo studies have suggested that the timing of systolic stretch can affect regional tension and external work development. In the present study, we measured the direct effects of systolic stretch timing on the magnitude of tension and external work development in isolated murine right ventricular papillary muscles. A servomotor was used to impose precisely timed stretches relative to electrical activation while a force transducer measured force output and strain was monitored using a charge-couple device camera and topical markers. Stretches taking place during peak intracellular Ca2+ statistically increased peak tension up to 270%, whereas external work due to stretches in this interval reached values of 500 J/m. An experimental analysis showed that time-varying elastance overestimated peak tension by 100% for stretches occurring after peak isometric tension. The addition of the force-velocity relation explained some effects of stretches occurring before the peak of the Ca2+ transient but had no effect in later stretches. An estimate of transient deactivation was measured by performing quick stretches to dissociate cross-bridges. The timing of transient deactivation explained the remaining differences between the model and experiment. These results suggest that stretch near the start of cardiac tension development substantially increases twitch tension and mechanical work production, whereas late stretches decrease external work. While the increased work can mostly be explained by the time-varying elastance of cardiac muscle, the decreased work in muscles stretched after the peak of the Ca2+ transient is largely due to myofilament deactivation.

Effects of angiotensin II and vasopressin on intracellular pH of glomerular mesangial cells

1988 ◽  
Vol 254 (6) ◽  
pp. F787-F794 ◽  
Author(s):  
M. B. Ganz ◽  
G. Boyarsky ◽  
W. F. Boron ◽  
R. B. Sterzel
Keyword(s):  
Angiotensin Ii ◽  
Intracellular Ph ◽  
Mesangial Cells ◽  
Ang Ii ◽  
Glomerular Mesangial Cells ◽  
Initial Value ◽  
Ph Change ◽  
Acid Load ◽  
Ethanesulfonic Acid ◽  
Acid Extrusion

We investigated changes in intracellular pH (pHi) of cultured rat glomerular mesangial cells (MCs) exposed to angiotensin II (ANG II) and arginine vasopressin (AVP). pHi of quiescent MCs, passage 2–5, and grown on glass cover slips, was assessed by spectrofluorometry using the pH-sensitive dye, 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF). The steady-state pHi of MCs in a pH 7.4, HCO3-free N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered solution was 7.10 +/- 0.02 (n = 68) and in a pH 7.4, HCO3-containing solution, was 7.23 +/- 0.03 (n = 47) (P less than 0.01). The pHi recovery following an NH+4-induced acid load was inhibited by removal of Na+ from the bath or by addition of the amiloride analogue, ethyl isopropyl amiloride (EIPA). These effects were observed in MCs bathed in HEPES- or in HCO3-buffered solutions, consistent with the action of a Na+-H+ exchanger. When cells were bathed in HEPES, a 10-min exposure to ANG II or AVP (10(-10) to 10(-6) M) caused early and transient acidification of MCs (maximal pH change was -0.10), followed by gradual alkalinization (maximal pHi change +0.15 above the initial value). The increase of pHi was dependent on the presence of Na+ in the bath and was inhibited by EIPA. In the presence of HCO3, ANG II or AVP induced merely a small gradual acidification of MCs (pHi change -0.05). These findings demonstrate that MCs utilize a Na+-H+ exchanger for acid extrusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II-induced endothelium-dependent relaxation of fowl aorta

1993 ◽  
Vol 264 (5) ◽  
pp. R903-R911 ◽  
Author(s):  
K. Hasegawa ◽  
H. Nishimura ◽  
M. C. Khosla
Keyword(s):  
Angiotensin Ii ◽  
Smooth Muscles ◽  
Isometric Tension ◽  
High Dose ◽  
Ang Ii ◽  
Bathing Medium ◽  
Calmodulin Antagonist ◽  
Endothelium Dependent Relaxation

In the domestic fowl, angiotensin II (ANG II) decreases blood pressure in vivo and causes endothelium-dependent relaxation of aortic smooth muscles in vitro. To characterize ANG II-induced vasorelaxation, we compared endothelium-dependent vasodilatory effects of [Asp1,Val5]-ANG II (fowl ANG II) and acetylcholine (ACh) with the endothelium-independent vasorelaxing effect of sodium nitroprusside (SNP) on isometric tension of fowl aortic rings. Hemoglobin (Hb), gossypol, and N omega-nitro-L-arginine methyl ester (L-NAME), inhibitors for endothelium-derived relaxing factor (EDRF) in mammalian blood vessels, partially inhibited vasorelaxation induced by ANG II and ACh in fowl. Hb also markedly attenuated SNP-induced vasorelaxation, but not 8-bromoguanosine 3',5'-cyclic monophosphate-induced relaxation. 3,4,5-Trimethoxybenzoic acid 8-(diethylamino)octyl ester hydrochloride (TMB-8) or the removal of Ca2+ from the bathing medium attenuated the ACh-induced relaxation but did not significantly reduce vasorelaxation induced by ANG II or SNP. In the zero Ca2+ medium, aortic rings showed tachyphylaxis to ACh, while ANG II caused tachyphylaxis regardless of the presence or absence of external Ca2+. Furthermore, pretreatment of the ring with a high dose of ACh abolished the vasorelaxation response to ANG II, suggesting that ACh and ANG II may share a common Ca2+ pool. Calmidazolium, a calmodulin antagonist, abolished the vasorelaxation induced by ANG II and ACh but not that by SNP. Comparison of the vasodilatory effects of several ANG II analogues on fowl aortic rings showed an approximate potency order of [Asp1,Val5]-ANG II = [Asp1,Ile5]-ANG II > [Asn1,Ile5]-ANG II = [Sar1,Ile5]-ANG II > [Val5]-ANG III.(ABSTRACT TRUNCATED AT 250 WORDS)

Arterial muscle contraction and potassium movement in vitro

1962 ◽  
Vol 202 (1) ◽  
pp. 17-20 ◽  
Author(s):  
Verle E. Headings ◽  
Paul A. Rondell
Keyword(s):  
Muscle Contraction ◽  
Potassium Content ◽  
Physiological Salt Solution ◽  
Time Intervals ◽  
Potassium Efflux ◽  
Tension Development ◽  
Maximum Loss ◽  
Indirect Action

Rings of dog carotid 2–3 mm in width were placed in a physiological salt solution, equilibrated, under 1 g of tension, at 37 C for 2 hr and were then stimulated electrically or with epinephrine, or norepinephrine. Tension development was recorded. At given time intervals following stimulation, rings were removed and analyzed for potassium content. Maximum loss of potassium, averaging 10% of original content, occurred 5–15 min after electrical stimulation. After stimulation by either epinephrine or norepinephrine a consistent slight increase in potassium content occurred. Thus net potassium efflux (and membrane depolarization?) is not an essential part of vascular muscle contraction in response to catecholamine stimulation. The reported loss of potassium from artery wall following catecholamine stimulation in vivo suggests another, perhaps indirect, action of these agents.

Na(+)-H+ and Na(+)-dependent Cl(-)-HCO3- exchange control pHi in vascular smooth muscle

1990 ◽  
Vol 259 (1) ◽  
pp. C134-C143 ◽  
Author(s):  
A. M. Kahn ◽  
E. J. Cragoe ◽  
J. C. Allen ◽  
R. D. Halligan ◽  
H. Shelat
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Transport Systems ◽  
Disulfonic Acid ◽  
Physiological Salt Solution ◽  
Physiological Conditions ◽  
Acid Load ◽  
22Na Uptake ◽  
Sarcolemmal Transport ◽  
Exchange Control

The mechanisms that control intracellular pH (pHi) in vascular smooth muscle are not fully understood. These studies were performed to determine the identity and relative importance of the sarcolemmal transport systems that mediate net acid efflux in primary cultured vascular smooth muscle cells from canine femoral artery. In HEPES- or HCO3(-)-buffered physiological salt solution (HEPES-PSS, HCO3(-)-PSS), recovery from an acute acid load was totally dependent on external Na+. 5-[N-ethyl-N-isopropyl]amiloride (EIPA, 50 microM) inhibited pHi recovery 100 and 68% in HEPES-PSS and HCO3(-)-PSS, respectively. EIPA-insensitive pHi recovery in HCO3(-)-PSS was inhibited 48% by 4,4'-diisothyocyanostilbene-2,2'-disulfonic acid (DIDS). An outwardly directed H+ gradient stimulated amiloride-sensitive 22Na+ uptake, and an inwardly directed HCO3- gradient stimulated amiloride-insensitive 22Na+ uptake. The latter was inhibited by DIDS or prior depletion of cell Cl-. In HEPES-PSS, resting pHi was 7.17 +/- 0.03, was not affected by DIDS, but was lowered by EIPA or by removing extracellular Na+. In HCO3(-)-PSS, resting pHi was 7.25 +/- 0.02 (P less than 0.05) and was not affected by EIPA. Removing extracellular Na+ in the presence of EIPA decreased pHi in HCO3(-)-PSS but not in HEPES-PSS. DIDS lowered resting pHi in HCO3(-)-PSS, after which EIPA further lowered pHi. We conclude that acid efflux from these cells is mediated by a Na(+)-H+ exchanger and a Na(+)-dependent Cl(-)-HCO3- exchanger. In HEPES-PSS, acid efflux via the Na(+)-H+ exchanger maintains resting pHi. In HCO3(-)-PSS, additional acid efflux via the Na(+)-dependent Cl(-)-HCO3- exchanger results in a higher pHi. Although the Na(+)-H+ exchanger is primarily responsible for acid efflux after an acute acid load, the Na(+)-dependent Cl(-)-HCO3- exchanger is responsible for acid efflux under physiological conditions.

pH regulation in hepatoma cells: roles for Na-H exchange, Cl-HCO3 exchange, and Na-HCO3 cotransport

1989 ◽  
Vol 257 (3) ◽  
pp. G317-G327 ◽  
Author(s):  
W. H. Weintraub ◽  
T. E. Machen
Keyword(s):  
Buffer Capacity ◽  
Ph Regulation ◽  
Hepatoma Cell ◽  
Disulfonic Acid ◽  
Hepatoma Cell Line ◽  
Free Treatment ◽  
Acid Load ◽  
Flow Through ◽  
Ethanesulfonic Acid ◽  
Acid Loading

Regulation of intracellular pH (pHi) was studied in Fu5, a rat hepatoma cell line that maintains a variety of differentiated functions. Microspectrofluorimetry of the pH-sensitive dye 2',7'-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF) was used to measure pHi in 10-15 cells growing on cover glasses that were mounted in a flow-through chamber on the stage of a microscope. In N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered solutions, pHi was 7.14, and intrinsic buffer capacity was inversely related to pHi. Amiloride (0.1 mM) caused pHi to decrease by 0.33 pH units in 4 min. Recovery from an acid load (using either NH4 prepulse technique or Na-free solutions) was completely blocked by amiloride. In HCO3-CO2-buffered solutions, pHi was 7.15, and buffer capacity was relatively insensitive to pHi between pHi of 6.6 and 7.2. Amiloride caused pHi to decrease by only 0.09 units. Recovery from an acid load was Na dependent, occurred in Cl-free solutions, and was totally blocked by the combination of amiloride plus 0.5 mM dihydro-4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (H2DIDS); recovery occurred when either amiloride or H2DIDS was removed. Removal of external Cl caused a rapid, H2DIDS-blockable alkalinization that was faster in HCO3-CO2 than in HEPES. The apparent Km for Clout for relaxation of Cl-free alkalinization was 4.5 mM. Rate of HCO3 transport during Cl-free treatment increased at alkaline resting pHi. It is concluded that Fu5 cells have two Na-dependent base-loading mechanisms and an acid-loading Cl-HCO3 exchanger. In solutions containing HCO3-CO2, the Na-H exchanger accounts for approximately 40% of recovery from an acid load, and a Na-HCO3 cotransporter accounts for the remainder. Recovery from an alkaline load appears to occur through the activity of the Cl-HCO3 exchanger.

Influence of aldosterone on isometric tension development in the rat heart

1963 ◽  
Vol 204 (6) ◽  
pp. 1001-1004 ◽  
Author(s):  
William C. Ullrick ◽  
Robert L. Hazelwood
Keyword(s):  
Heart Muscle ◽  
Rat Heart ◽  
Isolated Rat Heart ◽  
Isometric Tension ◽  
Muscle Performance ◽  
Tension Development ◽  
Tension Time ◽  
Isometric Twitch

Isometric twitch curves were recorded from preparations of rat heart trabeculae carneae removed from normal or adrenalectomized animals, or from normal animals injected for 3 days with saline or with 3 µg/day of d-aldosterone. Preparations from normal and from adrenalectomized animals were stimulated for 60 min in aldosterone-free Ringer's solution, or for 30 min in aldosterone-free Ringer's followed by 30 min in Ringer's containing 3 x 10–4 µg/ml of d-aldosterone. For a number of preparations from adrenalectomized animals the concentration of aldosterone was increased to 3 x 10–1 µg/ml. Regardless of treatment, all preparations were stimulated for a total of 1 hr at a rate of approximately 395/min; subsequently the recorded twitch curves were analyzed for peak tension development and for tension-time area. Although adrenalectomy tended to lower these variables of in vitro heart muscle performance, aldosterone, either administered in vivo or added directly to the isolated muscle bath, was without influence. It is concluded that aldosterone in the concentrations used does not alter the isometric tension characteristics of isolated rat heart muscle.

Endotoxemia-induced myocardial dysfunction is not associated with changes in myofilament Ca2+responsiveness

1998 ◽  
Vol 274 (2) ◽  
pp. H580-H590 ◽  
Author(s):  
Sherri L. Rigby ◽  
Polly A. Hofmann ◽  
Juming Zhong ◽  
H. Richard Adams ◽  
Leona J. Rubin
Keyword(s):  
Ventricular Myocytes ◽  
Contractile Function ◽  
Myocardial Dysfunction ◽  
Isometric Tension ◽  
Saline Control ◽  
Tension Development ◽  
Maximal Tension ◽  
Myocardial Contractile ◽  
The Relationship

Myocardial contractile function is depressed after onset of endotoxemia and is intrinsic to the ventricular myocyte. We tested the hypothesis that decreased Ca2+ responsiveness of the contractile myofilaments underlies this inotropic depression. Specifically, we evaluated the relationship between Ca2+ and unloaded cell shortening and isometric tension development of skinned guinea pig ventricular myocytes. Myocytes were isolated 4 h after intraperitoneal injection of 4 mg/kg Escherichia colilipopolysaccharide (LPS) or saline (control; Ctl). Myofilament Ca2+ responsiveness assessed by image analysis of shortening of skinned myocytes at pH 7.0 was not different between Ctl [pCa value that resulted in half-maximal shortening (pCa50): 5.78 ± 0.04] and LPS (pCa50: 5.72 ± 0.02). Similarly, myofilament Ca2+ responsiveness measured by isometric tension of skinned myocytes was not different between Ctl (pCa50: 5.73 ± 0.02) and LPS (pCa50: 5.76 ± 0.02). Maximal tension generated by LPS myocytes (2.89 ± 0.23 g/mm2) was significantly less ( P < 0.05) than Ctl (3.75 ± 0.34 g/mm2). However, when myocytes were isolated and skinned in the presence of protease inhibitors, maximal tension generated by LPS myocytes (3.53 ± 0.98 g/mm2) was similar to Ctl (3.01 ± 0.80 g/mm2). We conclude that in vivo administration of LPS resulting in endotoxemia without shock does not alter myofilament Ca2+ responsiveness of ventricular myocytes. Rather, reduced contractility is more likely a result of decreased Ca2+ availability because systolic Ca2+ transients of fura 2-loaded LPS myocytes were significantly decreased ( P < 0.05) compared with Ctl myocytes.

Luminal flow rate regulates proximal tubule H-HCO3 transporters

1992 ◽  
Vol 262 (1) ◽  
pp. F47-F54 ◽  
Author(s):  
P. A. Preisig
Keyword(s):  
Proximal Tubule ◽  
Flow Rate ◽  
Initial Rate ◽  
Apical Membrane ◽  
Basolateral Membrane ◽  
Rate Dependence ◽  
Unstirred Layer ◽  
Disulfonic Acid ◽  
Luminal Flow

In vivo microperfusion was used to examine the mechanism of luminal flow rate dependence of proximal tubule acidification. Luminal flow rate was acutely changed between 5 and 40 nl/min, while luminal and peritubular capillary composition were held constant. With inhibition of basolateral membrane base transport by peritubular 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), cell pH (pHi) provides a sensitive index of apical membrane H secretory activity. At a luminal perfusate [HCO3] of 25 mM, progressive increases in luminal flow rate (5----15----25----40 nl/min) caused progressive increases in pHi. This effect was of a smaller magnitude with a luminal perfusate [HCO3] of 60 mM and was further decreased at a luminal perfusate [HCO3] of 100 mM. This pattern of diminished flow rate dependence at higher luminal [HCO3] is consistent with the presence of a luminal unstirred layer, whose composition can be modified by luminal flow rate. The activity of the apical membrane Na-H antiporter, assayed as the initial rate of pHi recovery from an acid load in the presence of peritubular DIDS, was faster at 40 compared with 5 nl/min. Basolateral membrane Na-3HCO3 symporter activity, assayed as the initial rate of pHi recovery from an alkali load in the absence of luminal and peritubular chloride, was faster at 40 compared with 5 nl/min. This effect was eliminated by luminal amiloride, suggesting an indirect effect of flow mediated by changes in pHi secondary to flow rate-dependent changes in apical membrane Na-H antiporter activity. In summary, increases in luminal flow rate directly increase apical membrane H secretion, possibly by modification of a luminal unstirred layer.(ABSTRACT TRUNCATED AT 250 WORDS)

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