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D Tillotson

Publications and source records attributed to D Tillotson.

11 recordsLinked to original sources

Oscillations in cytosolic free Ca2+, oxygen consumption, and insulin secretion in glucose-stimulated rat pancreatic islets.

Insulin secretion in the intact organism, and by the perfused pancreas and groups of isolated perifused islets, is pulsatile. We have proposed a metabolic model of glucose-induced insulin secretion in which oscillations in the ATP/ADP ratio drive alterations in metabolic and electrical events that lead to insulin release. A key prediction of our model is that metabolically driven Ca2+ oscillations will also occur. Using the fluorescent Ca2+ probe, fura 2, digital image analysis, and sensitive O2 electrodes, we investigated cytosolic free Ca2+ responses and O2 consumption in perifused rat islets that had been maintained in culture for 1-4 days. We found that elevated ambient glucose increased the average cytosolic free Ca2+ level, the ATP/ADP ratio, and oxygen consumption, as previously found in freshly isolated islets. Oscillatory patterns were obtained for Ca2+, O2 consumption, and insulin secretion in the presence of 10 and 20 mM glucose. Very low amplitude oscillations in cytosolic free Ca2+ were observed at 3 mM nonstimulatory glucose levels. Evaluation of the Ca2+ responses of a large series of individual islets, monitored by digital image analysis and perifused at both 3 and 10 mM glucose, indicated that the rise in glucose concentration caused more than a doubling of the average cytosolic free Ca2+ value and a 4-fold increase in the amplitude of the oscillations with little change in period. The pattern of Ca2+ change within the islets was consistent with recruitment of responding cells. The coexistence of oscillations with similar periods in insulin secretion, oxygen consumption, and cytosolic free Ca2+ is consistent with the model of metabolically driven pulsatile insulin secretion.

Adenine Nucleotides

Effects of calculator use on arithmetic and conceptual skills of nursing students.

The purpose of this study was to determine the effect of the use of calculators on the responses of undergraduate nursing students to items on a dosage calculations examination. The study sought to determine if calculator use had any influence on both the arithmetic and conceptual skills associated with the solving of calculations problems. The population consisted of all students enrolled in a pharmacology course in a baccalaureate nursing program. An experimental repeated measures design was used in which all participants completed two parallel forms of a calculations examination, one form with a calculator and one form without a calculator. Findings of the study revealed that there was significant difference in both arithmetic and conceptual skills ability using a calculator. Use of a calculator was associated with improved arithmetic skills performance and diminished conceptual skills performance. A major factor underlying both arithmetic skills and conceptual skills performance, independent of the calculator effect, was the pre-admission arithmetic skills ability of the subject.

Concept Formation

Effects of Na+ and Ca2+ gradients on intracellular free Ca2+ in voltage-clamped Aplysia neurons.

Selected neurons of the abdominal ganglion of Aplysia californica were voltage-clamped and intracellular free Ca [( Ca2+]i) and Na [( Na+]i) concentrations were monitored with ion selective microelectrodes. Reducing [Na+]o from 500 mM (normal seawater, NSW) to 5 mM resulted in a decrease of the potential measured by the Ca electrode (VCa). Increasing [Ca2+]o from 10 to 50 mM increased [Ca2+]i two-fold, keeping [Ca2+]o at 50 mM and decreasing [Na+]o to 5 mM still led to a decrease in VCa. With 100 mM [Ca2+]o, which also increased [Ca2+]i, decreasing [Na+]o increased VCa in two of the eight cells tested. This indicates that in normal or moderately high resting [Ca2+]i, Ca2+ extrusion by Na/Ca exchange (forward mode) is not essential for [Ca2+]i buffering. [Na+]i was 12.9 +/- 3.6 mM (S.E.M., n = 7) in NSW; reducing [Na+]o to 5 mM decreased [Na+]i to 2.0 +/- 1.1 mM (S.E.M.). Keeping [Na+]o at 5 mM and increasing [Ca2+]o from 10 to 20 mM further decreased [Na+]i to about 1.0 mM, evidence of Na/Ca exchange operating in the reverse mode. Attempts to increase [Ca2+]i by bath application of the Ca ionophores A23187, X537A, ionomycin or ETH 1001 resulted in no measurable change of the resting [Ca2+]i. Application of Ouabain caused an apparent increase in [Ca2+]i in two of the six cells tested. In cells injected with the metallochromic indicator arsenazo III (AIII), the rate of the falling phase of the AIII absorbance increase, following a voltage-clamp pulse, was significantly slower in 5 mM [Na+]o. This indicates that in its forward mode Na-Ca exchange is active in clearing large submembrane increases in [Ca2+]i.

Animals

Ability of the Ca2+-selective microelectrodes to measure fast and local Ca2+ transients in nerve cells.

The ability of the Ca2+-selective microelectrode to measure fast Ca2+ transients intracellularly is reviewed. In vitro, Ca microelectrodes can respond to Ca2+ injections with time to peaks as small as 40 ms. We present methods to improve the dynamic response of Ca microelectrodes and to make Ca-buffered solutions in high ionic strength. Examples of measurements of intracellular free Ca2+ [( Ca2+]i) transients in Aplysia neurons and in Limulus photoreceptors are shown. To show the validity of those measurements, simultaneous recordings of the Arsenazo III (AIII) absorbance and of the Ca-selective electrode potential were made in voltage-clamped neurons of the abdominal ganglion of Aplysia californica. Pressure injection of AIII to a concentration of 300-500 microM induced a rise in resting [Ca2+]i; injection of higher [AIII] led to buffering of [Ca2+]i transients. Both techniques responded to changes in resting [Ca2+]i in the same direction except that AIII showed an increase in absorbance in 0 [Ca2+]o. Voltage-clamp pulses transiently increased both the AIII absorbance and the Ca2+ electrode potential. Reducing or increasing the driving force for Ca2+ entry changed the magnitude of both signals in the right direction. Examples of spatial localization of [Ca2+]i increases and Ca2+ gradients within the cytoplasm were demonstrated using the Ca electrode. The use of optical techniques to measure local [Ca2+]i changes is briefly reviewed.

Animals

The rate of diffusion of Ca2+ and Ba2+ in a nerve cell body.

A spectrophotometric method was developed to directly measure the diffusion rate of Ca2+ and some other ions in nerve cell bodies, using pulsed ionophoretic injections and an optical microprobe to record locally absorbance changes of the dye arsenazo III. We report here that Ca2+ and Ba2+ diffuse at approximately the same rate in nerve soma cytoplasm, having effective diffusion coefficients in the range of 7-12 X 10(-7) cm2/s, while identical measurements conducted in an electrolytic solution yielded values of 5.2 X 10(-6) cm2/s for Ca and 5.4 X 10(-6) cm2/s for Ba. The results are discussed in relation to the mechanisms that regulate the intracellular concentration of free Ca.

Animals

Inactivation of Ca conductance dependent on entry of Ca ions in molluscan neurons.

Inactivation of the Ca channel of Aplysia neurons was studied in the absence of potassium current in cells that were cesium-loaded with the aid of the ionophore nystatin. Inactivation was substantially decreased by methods that limited Ca entry. Depolarizations commensurate with the equilibrium potential of Ca resulted in minimal inactivation. Replacement of extracellular Ca by Ba also decreased inactivation. It is concluded that inactivation of the Ca channel is a function of the extent of Ca entry rather than membrane potential, thus differing fundamentally from the purely voltage-dependent mechanism for sodium inactivation.

Animals

Potassium activation associated with intraneuronal free calcium.

Relations between calcium entry and activation of a calcium-dependent outward current during depolarization were examined under voltage clamp in dorid giant neurons injected with the calcium-sensitive photoprotein aequorin. Activation kinetics and amplitude of the slow calcium-dependent component were both found to be related to the rate and extent of free calcium accumulation and to the electromotive force acting on potassium ions, independent of the calcium activation kinetics. This indicates that the activation of the calcium-dependent outward current is more closely related to the transient intracellular accumulation of free calcium ions than to the movement of calcium through the plasma membrane during depolarization.

Animals

Voltage-dependent facilitation of Ca2+ entry in voltage-clamped, aequorin-injected molluscan neurons.

Voltage-clamp experiments were performed on giant neurons of the nudibranch Anisodoris nobilis injected with the Ca-sensitive photoprotein, aequorin. Depolarization beyond -10 to +5 m V produced an aequorin signal, the amplitude of which depended on the extracellular Ca2+ concentration, the amplitude of the depolarization, and its duration. In paired pulse experiments, the amplitude of the aequorin signal produced in response to the second of two identical depolarizing pulses was larger than that produced during the first, resulting from an increased entry of Ca2+ during the second pulse. The increment in Ca conductance inferred from the augmented signal during the second pulse was independent of Ca2+ influx during the first pulse but, instead, was related to the amplitude and duration of the first pulse.

Aequorin