Effects of the intraluminal Ca load on the kinetics of 45Ca uptake and efflux in brain microsomes

1996 ◽  
Vol 271 (5) ◽  
pp. C1472-C1479 ◽  
Author(s):  
K. M. Wells ◽  
R. F. Abercrombie

Effects of increasing intraluminal Ca ([Ca]i) on the kinetics of rat brain microsomal uptake and efflux are reported here. Isolated rat brain microsomes accumulated 45Ca in an extravesicular free Ca ([Ca]o)- and ATP-dependent manner. Increased microsomal Ca load resulted in a decreased initial rate of 45Ca uptake and an increased tau, time to reach 63% of steady-state accumulation. Isolated rate brain microsomes lost 45Ca in a temperature- and [Ca]i-dependent manner. Whether preloaded with tracer 45Ca and either < or = 0.5 or 25 microM [Ca]o, the time constant of efflux was larger at 4 degrees C as compared with 37 degrees C. Additionally, increased microsomal Ca load resulted in a decreased time constant of 45Ca efflux. This shorter efflux time constant cannot explain the effect of [Ca]i on tau during uptake which was in fact longer for preloaded microsomes. Rather, these data suggest that, as Ca accumulates into unloaded microsomes, a steadily increasing [Ca]i slows unidirectional Ca influx (presumably by inhibiting the endoplasmic reticulum Ca pump) and enhances unidirectional Ca efflux, and that these combined effects ultimately shorten the time needed to achieve steady-state luminal [Ca]i.

1988 ◽  
Vol 8 (5) ◽  
pp. 1957-1969 ◽  
Author(s):  
R A Shapiro ◽  
D Herrick ◽  
R E Manrow ◽  
D Blinder ◽  
A Jacobson

As an approach to understanding the structures and mechanisms which determine mRNA decay rates, we have cloned and begun to characterize cDNAs which encode mRNAs representative of the stability extremes in the poly(A)+ RNA population of Dictyostelium discoideum amoebae. The cDNA clones were identified in a screening procedure which was based on the occurrence of poly(A) shortening during mRNA aging. mRNA half-lives were determined by hybridization of poly(A)+ RNA, isolated from cells labeled in a 32PO4 pulse-chase, to dots of excess cloned DNA. Individual mRNAs decayed with unique first-order decay rates ranging from 0.9 to 9.6 h, indicating that the complex decay kinetics of total poly(A)+ RNA in D. discoideum amoebae reflect the sum of the decay rates of individual mRNAs. Using specific probes derived from these cDNA clones, we have compared the sizes, extents of ribosome loading, and poly(A) tail lengths of stable, moderately stable, and unstable mRNAs. We found (i) no correlation between mRNA size and decay rate; (ii) no significant difference in the number of ribosomes per unit length of stable versus unstable mRNAs, and (iii) a general inverse relationship between mRNA decay rates and poly(A) tail lengths. Collectively, these observations indicate that mRNA decay in D. discoideum amoebae cannot be explained in terms of random nucleolytic events. The possibility that specific 3'-structural determinants can confer mRNA instability is suggested by a comparison of the labeling and turnover kinetics of different actin mRNAs. A correlation was observed between the steady-state percentage of a given mRNA found in polysomes and its degree of instability; i.e., unstable mRNAs were more efficiently recruited into polysomes than stable mRNAs. Since stable mRNAs are, on average, "older" than unstable mRNAs, this correlation may reflect a translational role for mRNA modifications that change in a time-dependent manner. Our previous studies have demonstrated both a time-dependent shortening and a possible translational role for the 3' poly(A) tracts of mRNA. We suggest, therefore, that the observed differences in the translational efficiency of stable and unstable mRNAs may, in part, be attributable to differences in steady-state poly(A) tail lengths.


1992 ◽  
Vol 73 (5) ◽  
pp. 1939-1945 ◽  
Author(s):  
E. M. Postlethwait ◽  
S. D. Langford ◽  
A. Bidani

We previously showed, during quasi-steady-state exposures, that the rate of inhaled NO2 uptake displays reaction-mediated characteristics (J. Appl. Physiol. 68: 594–603, 1990). In vitro kinetic studies of pulmonary epithelial lining fluid (ELF) demonstrated that NO2 interfacial transfer into ELF exhibits first-order kinetics with respect to NO2, attains [NO2]-dependent rate saturation, and is aqueous substrate dependent (J. Appl. Physiol. 71: 1502–1510, 1991). We have extended these observations by evaluating the kinetics of NO2 gas phase disappearance in isolated ventilating rat lungs. Transient exposures (2–3/lung at 25 degrees C) employed rebreathing (NO2-air) from a non-compliant continuously stirred closed chamber. We observed that 1) NO2 uptake rate is independent of exposure period, 2) NO2 gas phase disappearance exhibited first-order kinetics [initial rate (r*) saturation occurred when [NO2] > 11 ppm], 3) the mean effective rate constant (k*) for NO2 gas phase disappearance ([NO2] < or = 11 ppm, tidal volume = 2.3 ml, functional residual capacity = 4 ml, ventilation frequency = 50/min) was 83 +/- 5 ml/min, 4) with [NO2] < or = 11 ppm, k* and r* were proportional to tidal volume, and 5) NO2 fractional uptakes were constant across [NO2] (< or = 11 ppm) and tidal volumes but exceeded quasi-steady-state observations. Preliminary data indicate that this divergence may be related to the inspired PCO2. These results suggest that NO2 reactive uptake within rebreathing isolated lungs follows first-order kinetics and displays initial rate saturation, similar to isolated ELF.(ABSTRACT TRUNCATED AT 250 WORDS)


1999 ◽  
Vol 22 (3) ◽  
pp. 253-256 ◽  
Author(s):  
Shizuo NARIMATSU ◽  
Shigeo YAMAMOTO ◽  
Tomoni KOITABASHI ◽  
Rika KATO ◽  
Yasuhiro MASUBUCHI ◽  
...  

1997 ◽  
Vol 272 (4) ◽  
pp. C1087-C1098 ◽  
Author(s):  
E. E. Burmeister Getz ◽  
S. L. Lehman

The models of the sarcoplasmic reticulum (SR) Ca pump used to simulate Ca kinetics in muscle fibers are simple but inconsistent with data on Ca binding or steady-state uptake. We develop a model of the SR pump that is consistent with data on transient and steady-state Ca removal and has rate constants identified under near-physiological conditions. We also develop models of the other main Ca-binding proteins in skeletal muscle: troponin C and parvalbumin. These models are used to simulate Ca transients in cut fibers during and after depolarizing pulses. Simulations using the full SR pump model are contrasted with simulations using a Michaelis-Menten (MM) approximation to SR pump kinetics. The MM pump underestimates the amount of Ca released during depolarization, underestimates the initial rate of Ca binding by the pump, and overestimates the later rate of Ca pumping. These errors are due to fast initial binding by the SR pump, which is neglected in the MM approximation.


1988 ◽  
Vol 254 (5) ◽  
pp. C621-C627 ◽  
Author(s):  
T. W. Hurley

Energy-dependent subcellular Ca2+ sequestration was studied in the presence of ruthenium red using rat pancreatic acini, which had been permeabilized by exposure to medium nominally free of Ca2+. The initial rate of Ca2+ uptake (approximately 2,800 pmol.min-1.mg acinar protein-1) quickly slowed, and a mean steady-state Ca2+ content of approximately 3,000 pmol/mg was reached after 5-10 min of incubation at 37 degrees C. Ca2+ uptake was stimulated by submicromolar Ca2+ concentrations (K0.5 = 156 nM); required Mg2+-ATP (K0.5 = 0.78 mM) was greatest at a pH of 7.0 and was abolished by the Ca2+ ionophore A23187. Other nucleotide phosphates as well as p-nitrophenylphosphate were relatively poor substrates, supporting Ca2+ uptake at initial rates that were 6-14% of those measured in the presence of ATP. These results show that pancreatic acini permeabilized without detergents possess a nonmitochondrial Ca2+ transporting system not located in the plasma membrane but with the properties expected of a major regulator of acinar cytosolic Ca2+ concentration.


2005 ◽  
Vol 10 (3) ◽  
pp. 223-233 ◽  
Author(s):  
J. Kulys

Kinetics of biocatalytical synergistic reactions has been analyzed at non-stationary state (NSS) and at quasi steady state (QSS) conditions. The application to the model kinetic constants taken from the first type of the experiments shows that QSS can be established for the enzyme and the mediator at time less than 1 s. Therefore, the analytical solution of the initial rate (IR) may be produced at relevant to an experiment time, and the dependence of the IR on substrates concentration may be analyzed rather easy. The use of kinetic constants from the second type of reactions shows that QSS is formed for the enzyme but not for the mediator. For this reason the modeling of the synergistic process was performed by solving the ordinary differential equations (ODE). For this purpose the novel program KinFitSim (c) was used.


1998 ◽  
Vol 201 (14) ◽  
pp. 2103-2113 ◽  
Author(s):  
R Bräucker ◽  
A Murakami ◽  
K Ikegaya ◽  
K Yoshimura ◽  
K Takahashi ◽  
...  

The kinetics of gravitaxis and gravikinesis in Paramecium caudatum were investigated by employing (1) step transitions from normal gravity (1 g) to weightlessness (microgravity) and (2) turns of the experimental chambers from the horizontal to the vertical position at 1 g. The transition to microgravity left existing cell orientations unchanged. Relaxation of negative gravitaxis under microgravity took longer than 10 s and may be described by the time constant of the decay of orientation coefficients. Gravitaxis was started at 1 g by turning the experimental chamber from a horizontal to a vertical position. Gravitaxis activated rapidly during the turning procedure and relaxed to an intermediate level after the turning had stopped. Gravity-induced regulation of swimming speed (gravikinesis) at 1 g had reached a steady state after 1 min; at this point, gravikinesis counteracted the effects of sedimentation (negative gravikinesis). A step transition to microgravity initially reversed the sign of the gravikinesis (positive gravikinesis). The relaxation of this kinetic response was not completed during 10 s of microgravity. The data suggest that gravikinesis is functionally unrelated to gravitaxis and is strongly affected by the rate of change in acceleration. We present a model explaining why gravikinesis reverses sign upon the onset of a step from 1 g to microgravity.


ADMET & DMPK ◽  
2016 ◽  
Vol 4 (2) ◽  
pp. 91
Author(s):  
Alejandro Palacios ◽  
Javier Barberón ◽  
Patricio Leaden ◽  
Pedro Zeinsteger

<p class="ADMETabstracttext">In this study the effects of Calendula officinalis L. (Asteraceae) extract (CO) on the polyunsaturated fatty acid composition, chemiluminescence and unsaturation index of microsomes isolated from brain rat, are presented. After incubation of microsomes in an ascorbate (0.4 mM)-Fe<sup>2+</sup> (2.15 µM) system (180 min at 37 °C) it was observed that the total cpm/mg protein originated from light emission:chemiluminescence was lower in brain microsomes obtained from CO group compared to the control group (without extract supplementation). Moreover, it was observed that the addition of the extract reduced chemiluminescence -measured as total cpm- in a concentration dependent manner. The fatty acid composition of brain microsomes from control group was profoundly modified when subjected to non-enzymatic lipoperoxidation with a considerable decrease of arachidonic acid C20:4w6 and docosahexaenoic acid C22:6w3. As a consequence, the unsaturation index, a parameter based on the maximal rate of oxidation of specific fatty acids, was higher in the CO group compared to controls. The simultaneous analysis of unsaturation index, chemiluminescence and fatty acid composition indicate that CO may act as an antioxidant protecting rat brain microsomes from peroxidative damage.</p>


Sign in / Sign up

Export Citation Format

Share Document