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D C Eaton

Publications and source records attributed to D C Eaton.

102 records · Page 6Linked to original sources
PubMed1979

Correlation between macrophage intracellular electrical potentials and malignant melanoma growth in a murine model.

Peritoneal macrophages were collected from mice at varying periods after transplantation of an allogeneic malignant melanoma in the hind limb. The intracellular electrical potentials of these macrophages were measured and a correlation was found to exist between tumor growth measured by size and pathological examination, and the development of large negative intracellular potentials. We propose that this change in intracellular potential is correlated with changes in the immune system and may be triggered by membrane permeability changes possibly in response to calcium ions.

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PubMed1978

Sodium channel inactivation in squid axon is removed by high internal pH or tyrosine-specific reagents.

In squid axon, internal alkalinization from pH 7.1 to pH 10.2 results in a reversible decrease of the maximum inward current and the steady state sodium channel inactivation. Similar effects were observed after treatment of the axon with tetranitromethane or after iodination with lactoperoxidase. These results suggest that a tyrosine residue is an essential component of the inactivation process in this nerve.

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PubMed1978

Basolateral membrane potential of a tight epithelium: ionic diffusion and electrogenic pumps.

The contribution of specific ions to the conductance and potential of the basolateral membrane of the rabbit urinary bladder has been studied with both conventional and ion-specific microelectrode techniques. In addition, the possibility of an electrogenic active transport process located at the basolateral membrane was studied using the polyene antibiotic nystatin. The effect of ion-specific microelectrode impalement damage on intracellular ion activities was examined and a criterion set for acceptance or rejection of intracellular activity measurements. Using this criterion, we found (K+) = 72 mM and (Cl-) = 15.8 mM. Cl- but not K+ was in electrochemical equilibrium across the basolateral membrane. The selective permeability of the basolateral membrane was measured using microelectrodes, and the data analyzed using the Goldman, Hodgkin-Katz equation. The sodium to potassium permeability ratio (PNa/PK) was 0.044, and the chloride to potassium permeability ratio (PCl/PK) was 1.17. Since K+ was not in electrochemical equilibrium, intracellular (K+) is maintained by active metabolic processes, and the basolateral membrane potential is a diffusion potential with K+and C1- the most permeable ions. After depolarizing the basolateral membrane with high serosal potassium bathing solutions and eliminating the apical membrane as a rate limiting step for ion movement using the polyene antibiotic nystatin, we found that the addition of equal aliquots of NaCl to both solutions caused the basolateral membrane potential to hyperpolarize by up to 20mV (cell interior negative). This potential was reduced by 80% within 3 min of the addition of ouabain to the serosal solution. This hyperpolarization most probably represents a ouabain sensitive active transport process sensitive to intracellular Na+. An equivalent electrical circuit for Na+ transport across rabbit urinary bladder is derived, tested, and compared to previous results. This circuit is also used to predict the effects that microelectrode impalement damage will have on individual membrane potentials as well as time-dependent phenomena; e.g., effect of amiloride on apical and basolateral membrane potentials.

Animals↗
PubMed1977

Intracellular calcium and extra-retinal photoreception of Aplysia Giant neurons.

The early or "instantaneous" current-voltage relationship for the light-activated potassium current in Aplysia giant neurons was linear during the first second of illumination. However, the light current was greatly reduced or abolished by prolonged hyperpolarization. It was also greatly reduced by the injection of calcium EGTA buffers having calcium activities of 5.6 X 10(-8) M and simulated by injecting buffers with calcium activities of 2.8-5.6 X 10(-7) M. Removal of calcium from the extracellular fluid had no effect. Both the light- and calcium-activated outward potassium currents were reduced by tetraethylammonium (TEA) ions. The light current was not affected by substituting rubidium for potassium nor by substituting either lithium or Tris for sodium. The calcium-activated potassium current persisted when the neuron was cooled to 5 degrees C. However, the light response could no longer be elicited. Light hyperpolarizes Aplysia neurons probably by increasing intracellular calcium activity two-to six-fold which activates a membrane potassium conductance. Calcium levels appear to be restored within the cell and are energy dependent. The light-activated release of calcium is inhibited by cooling. The body wall of Aplysia transmits enough visible or 500 nm light to hyperpolarize some Aplysia giant neurons under ambient conditons. These neurons may be involved in the extraretinal light entrainment that occurs in Aplysia.

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PubMed1977

Effects of nystatin on membrane conductance and internal ion activities in Aplysia neurons.

Two methods were used to study effects of the antibiotics, nystatin, on giant neurons of Aplysia. In the first method the effects of various concentrations of nystatin on the current-voltage relationship were evaluated at a fixed time after exposure to the antibiotic using a two-microelectrode voltage clamp. Nystatin increased membrane conductance in a dose-dependent manner. The dose-response relation was very steep, with little or no effect below 15 mg/liter and an effect too large to measure at concentrations greater than 30 mg/liter. Upon return to antibiotic-free solution, membrane conductance returned to pre-treatment levels within 30 minutes. The second type of experiment involved use of ion-specific microelectrodes to measure changes of intracellular univalent ion activities which attended the nystatin-induced permeability. Nystatin-induced permeability changes mainly involved univalent cations, but Cl permeability was also increased. Nystatin may therefore be used to selectively rearrange the internal ionic milieu to study the effect of such a change on membrane transport or electrical properties.

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PubMed1977

Nystatin as a probe for investigating the electrical properties of a tight epithelium.

We show how the antibiotic nystatin may be used in conjunction with microelectrodes to resolve transepithelial conductance Gt into its components: Ga, apical membrane conductance; Gbl, basolateral membrane conductance; and Gj, junctional conductance. Mucosal addition of nystatin to rabbit urinary bladder in Na+-containing solutions caused Gt to increase severalfold to ca. 460 micrometerho/muF, and caused the transepithelial voltage Vt to approach +50 mV regardless of its initial value. From measurements of Gt and the voltage-divider ratio as a function of time after addition or removal of nystatin, values for Ga, Gbl, and Gj of untreated bladder could be obtained. Nystatin proved to have no direct effect on Gbl or Gj but to increase Ga by about two orders of magnitude, so that the basolateral membrane then provided almost all of the electrical resistance in the transcellular pathway. The nystatin channel in the apical membrane was more permeable to cations than to anions. The dose-response curve for nystatin had a slope of 4.6. Use of nystatin permitted assessment of whether microelectrode impalement introduced a significant shunt conductance into the untreated apical membrane, with the conclusion that such a shunt was negligible in the present experiments. Nystatin caused a hyperpolarization of the basolateral membrane potential in Na+-containing solutions. This may indicate that the Na+ pump in this membrane is electrogenic.

Animals↗
PubMed1976

The mechanism of Na+ transport by rabbit urinary bladder.

The mechanism of Na+ transport in rabbit urinary bladder has been studied by microelectrode techniques. Of the three layers of epithelium, the apical layer contains virtually all the transepithelial resistance. There is radial cell-to-cell coupling within this layer, but there is no detectable transverse coupling between layers. Cell coupling is apparently interrupted by intracellular injection of depolarizing current. The cell interiors are electrically negative to the bathing solutions, but the apical membrane of the apical layer depolarizes with increasing Isc. Voltage scanning detects no current sinks at the cell junctions or elsewhere. The voltage-divider ratio, alpha, (ratio of resistance of apical cell membrane, Ralpha, to basolateral cell membrane, Rb) decreases from 30 to 0.5 with increasing Isc, because of the transport-related conductance pathway in the apical membrane. Changes in effective transepithelial capacitance with Isc are predicted and possibly observed. The transepithelial resistance, Rt, has been resolved into Ra, Rb, and the junctional resistance, Rj, by four different methods: cable analysis, resistance of uncoupled cells, measurements of pairs of (Rt, alpha) values in the same bladder at different transport rates, and the relation between Rt and Isc and between alpha and Isc. Rj proves to be effectively infinite (nominally 300 k omega muF) and independent of Isc, and Ra decreases from 154 to 4 omega muF with increasing Isc. In the resulting model of Na+ transport in "tight" epithelia, the apical membrane contains an amiloride-inhibited and Ca++-inhibited conductance pathway for Na+ entry; the basolateral membrane contains a Na+--K+-activated ATPase that extrudes Na+; intracellular (Na+) may exert negative feedback on apical membrane conductance; and aldosterone acts to stimulate Na+ entry at the apical membrane via the amiloride-sensitive pathway.

Animals↗
PubMed1975

Ionic permeabilities of an Aplysia giant neuron.

In a giant neuron of Aplysia californica, permeabilities and conductances obtained by measuring net fluxes of Na+, K+ and Cl-minus with ion-specific microelectrodes were compared with those obtained by measuring transmembrane current and potential changes when the three ions were varied in the external solution. Net fluxes were measured with ion-specific microelectrodes, after blocking metabolic processes, thus allowing movement of ions down their electrochemical gradients. Premeabilities and conductances obtained from the "chemical" measurements (i.e., ion-specific electrodes) were generally comparable to the values obtained from "electrical" measurements (i.e., ion-specific electrodes) were generally comparable to the values obtained from "electrical" measurements. Where discrepancies occurred, they could be explained by showing that some of the assumptions necessary to use the "electrical" method were not quantitatively true in this system. The absolute magnitudes of the permeabilities are significantly less than those found in many axonal preparations. There is also a relatively high PNa/PK ratio. The selectivity of the membrane against ions such as Tris" and MeSO3-minus is not good, Tris+ being nearly as permeable as Na+ and MeSO3-minus about one-half as permeables as Cl-minus. These properties may be characteristic of somal membranes.

Animals↗
PubMed1974

Membrane currents carried by Ca, Sr, and Ba in barnacle muscle fiber during voltage clamp.

Membrane currents associated with voltage clamp of the giant muscle fibers of a barnacle, Balanus nubilus, were analyzed in terms of currents of the Ca and K channels. Although the activation of the K channel occurs more slowly than that of the Ca channel, both currents show a significant temporal overlap. The currents carried by Ca(++), Sr(++), or Ba(++) through the Ca channel were compared under the conditions at which this overlap was the least. When only one divalent cation is present in the solution, Ba(++) carries more current than Ca(++) or Sr(++) and the sequence of the current is Ba > Sr approximately Ca. When the external solution contains a relatively high concentration of Co(++), which is a blocking agent for the Ca channel, inversion of the sequence occurs, to Ca > Sr > Ba. This is due to the fact that the blocking effect differs depending on which ion carries current through the Ca channel. The Ba current is most sensitive and the Ca current is least affected. Ba suppresses the current of the K channel, independently of its current-carrying function through the Ca channel.

Animals↗
PubMed1972

Potassium ion accumulation near a pace-making cell of Aplysia.

1. A delayed current decrease associated with prolonged depolarization was studied in R(15) (the parabolic burster) of Aplysia by using intracellular recording and voltage clamp techniques.2. For long duration command pulses (3 sec), the outward current shows a delayed decrease. The current goes from a maximum near 100 msec and falls until a steady-state outward current is reached between 1.5 and 2.5 sec after the beginning of the command step. This final steady-state current is usually only about 20-30% of the peak outward current.3. Double step voltage clamps show that this current decrease is associated with a large shift of e.m.f. Measurements of conductance, on the other hand, fail to show any significant difference in conductance associated with peak and steady-state currents.4. From the results of application of high K(+) ringer, the conclusion is reached that this shift in e.m.f. is due to an accumulation of K(+) near the exterior cell membrane. Several other experiments exclude the possibility of either metabolic events or compensating conductance changes producing the phenomenon.5. The location of the accumulation is considered on the basis of anatomical studies. It is concluded that the accumulation takes place in the extensive infoldings found in cells like R(15). An explanation of the difference in delayed current decrease between pace-makers and non-pace-makers is suggested, since the pace-makers apparently have more extensive invaginations than the non-pace-makers. This suggestion is lent support by measurements of capitance and current density.

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