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D H Ross

Publications and source records attributed to D H Ross.

At least 37 records · Page 2Linked to original sources
PubMed1985

Use of mitochondrial inhibitors to differentiate kinetic properties of the ATP-dependent Ca2+ uptake system in synaptic membranes.

Synaptosomal membranes accumulate 3-6 times more Ca2+ in the presence of ATP (50-1000 microM) than basal Ca2+ accumulation (-ATP). The location of this Ca2+ accumulation appears to reside on the cytosolic face of the synaptosome since lysed synaptosomes accumulate 4-times more Ca2+ than intact synaptosomes. The inclusion of mitochondrial inhibitors, oligomycin (0.7 micrograms/ml), sodium azide (100 microM) and dinitrophenol (100 microM) differentiate mitochondrial from nonmitochondrial Ca2+ accumulation under conditions that are [Ca2+]- and ATP-dependent. In the presence of low concentrations of ATP (less than 150 microM) and Ca2+ free (2.5 or 6.8 microM), Ca2+ accumulation occurs as one process in both lysed synaptosomal membranes and purified synaptic plasma membranes in the presence and/or absence of MI. When ATP levels are increased (greater than 200 microM), Ca2+ accumulation process remains independent of the presence of mitochondrial inhibitors when Ca2+ free = 2.5 microM. When Ca2+ free is increased to 6.8 microM, mitochondrial inhibitors differentiate mitochondrial from nonmitochondrial accumulation. These studies suggest that optimal conditions for the measurement of Ca2+ accumulating mechanisms in synaptosomal membranes depend on both [Ca2+] and ATP. Use of these assay conditions provide evidence that ATP-dependent Ca2+ uptake may be a viable mechanism for the regulation of synaptosomal Ca2+ levels.

Adenosine Triphosphate↗
PubMed1985

The effects of lubrol WX on brain membrane Ca2+/Mg2+ ATPase and ATP-dependent Ca2+ uptake activity following acute and chronic ethanol.

Acute administration of ethanol (2.5 gm/kg, i.p.) to rats inhibits the cytosolic buffering of Ca2+ in nerve terminals. Ca2+ ATPase and ATP-dependent Ca2+ uptake are both inhibited 30 min after a single dose of ethanol. Chronic ethanol administration (6%, 14 days) did not inhibit Ca2+ ATPase but significantly stimulated ATP-dependent Ca2+ uptake. Lubrol WX treatment of acute ethanolic membranes reverses the inhibition of Ca2+ ATPase seen following ethanol. Lubrol WX treatment of chronic ethanolic membranes prevents the increase in ATP-dependent Ca2+ uptake seen in ethanolic membranes. Both acute and chronic ethanol-induced changes in Ca2+ transport within nerve terminals may involve lipid-dependent parameters of the membrane which may underlie neuronal adaptation.

Adenosine Triphosphate↗
PubMed1985

Substrate requirements and subcellular distribution of calcium transport activities in brain membranes.

Ca2+ transport activity in synaptosomal membranes has been identified as having two major components: Ca2+-stimulated ATP hydrolysis and ATP-dependent CA2+ uptake. Both processes exhibit similar affinities for Ca2+ and operate maximally under identical buffer conditions. Subcellular fractionation studies revealed the Ca2+/Mg2+ ATPase and ATP-dependent CA2+ uptake activities to be highest in synaptic plasma membrane fractions 1 and 2, with lesser activity in synaptic vesicles and mitochondria. Progressive treatment with Triton X-100 activated, then decreased Ca2+/Mg2+ ATPase, Mg2+ ATPase and Ca2+ ATPase. ATP-dependent Ca2+ uptake was progressively decreased by similar treatment with Triton X-100. These studies illustrate that Ca2+ ATPase and ATP-dependent Ca2+ uptake may provide two important mechanisms for buffering of cytosolic Ca2+ at the nerve terminal. These systems may function to rapidly sequester cytosolic Ca2+ following a rise during depolarization and then extrude Ca2+ from the terminal against a concentration gradient. This regulation of cytosolic Ca2+, represented by two processes of the type seen in other plasma membranes, may play critical roles in calcium homeostasis in nerve cells.

Adenosine Triphosphate↗
PubMed1985

Dibutyryl-cyclic GMP stimulation of Ca2+ -ATPase activity in rat brain synaptic membranes.

The effects of dibutyryl cyclic AMP (db-cAMP) and dibutyryl cyclic GMP (db-cGMP) were tested on Ca2+-ATPase, Mg2+-ATPase, and (Ca2+ + Mg2+)-ATPase activities in lysed synaptosomes prepared from whole rat brains (minus cerebellum). At concentrations from 0.1 to 2.0 mM, db-cGMP produced a selective, concentration-dependent increase in Ca2+-ATPase activity. Both db-cGMP and db-cAMP slightly reduced Mg2+-ATPase activity, whereas neither compound had concentration-dependent effects on (Ca2+ + Mg2+)-ATPase activity. These findings suggest that the Mg2+-independent, Ca2+-ATPase activity in rat brain is regulated by a cyclic GMP-dependent process. Further, the data provide evidence that the Ca2+-ATPase activity in lysed synaptosomal membranes represents an enzyme that is distinguishable from both the Mg2+ -and (Ca2+ + Mg2+)-ATPase.

Adenosine Triphosphatases↗
PubMed1983

Effects of in vivo ethanol administration on Ca2+/Mg2+ ATPase and ATP-dependent Ca2+ uptake activity in synaptosomal membranes.

Acute administration of ethanol (4 g/kg, i.p.) to mice inhibits the sequestration of calcium into endoplasmic reticulum-like organelles in synaptosomal membranes. Ethanol administration inhibits both Ca2+-stimulated adenosine triphosphate hydrolysis and ATP-dependent calcium uptake in the vesicles at time of loss of righting reflex. At recovery of righting reflex, the Ca2+-ATPase activity returns to normal levels, while the ATP-dependent uptake remains inhibited. The effect of ethanol is specific for the sequestration (active transport) of calcium since calcium binding to synaptic membranes is not altered. Alteration in mechanisms responsible for synaptosomal buffering of cytosolic Ca2+ levels by in vivo ethanol may contribute to altered transmitter release rates following ethanol administration.

Adenosine Triphosphate↗
PubMed1983

Calmodulin stimulation of Ca2+-dependent ATP hydrolysis and ATP-dependent Ca2+ transport in synaptic membranes.

We report here characterization of calmodulin-stimulated Ca2+ transport activities in synaptic plasma membranes (SPM). The calcium transport activity consists of a Ca2+-stimulated, Mg2+-dependent ATP hydrolysis coupled with ATP-dependent Ca2+ uptake into membraneous sacs on the cytosolic face of the synaptosomal membrane. These transport activities have been found in synaptosomal subfractions to be located primarily in SPM-1 and SPM-2. Both Ca2+-ATPase and ATP-dependent Ca2+ uptake require calmodulin for maximal activity (KCm for ATPase = 60 nM; KCm for uptake = 50 nM). In the reconstituted membrane system, KCa was found to be 0.8 microM for Ca2+-ATPase and 0.4 microM for Ca2+ uptake. These results demonstrate for the first time the calmodulin requirements for the Ca2+ pump in SPM when Ca2+ ATPase and Ca2+ uptake are assayed under functionally coupled conditions. They suggest that calmodulin association with the membrane calcium pump is regulated by the level of free Ca2+ in the cytoplasm. The activation by calmodulin, in turn, regulates the cytosolic Ca2+ levels in a feedback process. These studies expand the calmodulin hypothesis of synaptic transmission to include activation of a high-affinity Ca2+ + Mg2+ ATPase as a regulator for cytosolic Ca2+.

Animals↗
PubMed1982

Kinetic characterization and substrate requirement for the Ca2+ uptake system in platelet membrane.

An ATP-dependent mechanism for Ca2+ uptake in human platelet membrane fractions has been identified and characterized. Ca2+ uptake into a membrane fraction is shown to be stimulated at low concentrations of ATP and Ca2+ and to require magnesium ions. Initial rate kinetics, using Eadie-Scatchard analysis, indicated a single class of calcium uptake sites in the presence of ATP, with a Kd for free [Ca2+] of 0.145 microM. Ca2+ uptake in the presence of several ATP concentrations demonstrates that ATP binds to at least two sites, representing high and low affinities of 3.21 and 80.1 microM, respectively. The neuroleptic drug fluphenazine inhibited ATP-stimulated calcium uptake (IC50 = 55 microM), suggesting this ATP-dependent Ca2+ uptake system may provide a useful ion-transport model with which to study neuroleptic therapy in humans.

Adenosine Triphosphate↗
PubMed1981

Radiation and brain calcium: a review and critique.

Serious controversy pervades the scientific study of radio-frequency (RF) radiation and its biological effects. The issues range broadly from international differences in safe exposure standards to questions pertaining to the neurological symptoms purportedly induced by electromagnetic radiation. In a more specialized vein, there is great concern in the discipline about the influence of different sources of radiation on the activity of calcium in the brain. A principal and very realistic reason for this concern stems from the pivotal importance of calcium ions in the normal functioning of the brain in all of its myriad complexity. The purpose of the review is to critically evaluate from an unbiased and "non-involved" viewpoint the major findings on the possible interaction between calcium ions and various radiation sources. Background information is also considered as it relates even indirectly to hypothetical mechanisms that might be used to explain any possible shift in Ca++ ion kinetics. Finally, an inclusive critique is presented which deals with the bench-top methods and strategy used in the conduct of calcium-radiation experiments.

Animals↗
PubMed1981

Kinetic characterization of Ca2+ transport in synaptic membranes.

Lysed synaptosomal membranes were prepared from brain cortices of HA/ICR Swiss mice, and the ATP-stimulated Ca2+ uptake, Ca2+-stimulated Mg2+-dependent ATPase activity, and the Ca2+-stimulated acyl phosphorylation of these membranes were studied. The Km values for free calcium concentrations ([Ca2+]f) for these processes were 0.50 microM, 0.40 microM, and 0.31 microM, respectively. Two kinetically distinct binding sites for ATP were observed for the ATP-stimulated Ca2+ uptake and the Ca2+-stimulated Mg2+-ATPase activity. The high-affinity Km values for ATP for these two processes were 16.3 microM and 28 microM, respectively. These results indicate that the processes studied operate in similar physiological concentration ranges for the substrates [Ca2+]f and ATP under identical assay conditions and, further, that these processes may be functionally coupled in the membrane.

Adenosine Triphosphatases↗
PubMed1979

Nerve cell calcium as a messenger for opiate and endorphin actions.

The studies outlined in this chapter provide convincing evidence for the role of calcium in pharmacological and biochemical actions of opiates on the central nervous system. The ability of the synaptosomes to accumulate excess calcium during narcotic exposure represents a singular major example of cellular adaptation at the level of the membrane. These findings together with the possibility that CDR and calcium may mediate a number of processes involving neurotransmitter activity provide many avenues for the future study of opiates and other central nervous system depressants.

Animals↗
PubMed1979

Correlation of systemic and biochemical effects of PBB with behavioral effects.

Rats were given PBB orally at 1, 3, and 6 mg/kg (or vehicle as control) daily for 20 days. Some animals were sacrificed immediately while the food intake of remaining animals was limited to attain and maintain 80% of normal body weight. No effect of PBB upon body weight was observed for any of the dose levels employed. Immediately after dosing, liver/body weight ratios were 110% of controls for the 1 mg/kg group and 152% for the 3 and 6 mg/kg groups; after weight reduction for 2-6 months liver/body weight ratios for all 3 dose groups were 160-170% of controls. In the absence of body fat, most tissues exhibited dose-dependent retention of PBB 2-6 months after dosing with highest levels in liver followed by kidney. In 6 mg/kg rats weight-reduced for 6 months, liver AHH activity was 613% of controls; in rats sacrificed immediately after dosing, liver AHH activity was dose-related but appeared to reach maximal value at the 3 mg/kg dose. Both calcium binding to synaptic plasma membranes and calcium uptake by intact synaptosomes was significantly reduced in the brains of 1 mg/kg PBB rats, but not affected in preparations from 3 and 6 mg/kg animals.

Animals↗