Biomedical subjects
Publications and source records attributed to R Eckert.
Sublethally irradiated AB/Bln mice (500 cGy) were treated with splenotritin (fragment 32-34 of the splenic hormone splenin) and tested for their capability of producing antibodies against target sheep red blood cells. In x-ray plus splenotritin treated mice antibody forming cells were found earlier and in a higher frequency than in mice treated with x-rays alone.
Outward tail currents measured in Aplysia neurones after termination of depolarizing voltage-clamp pulses consist of rapidly decaying voltage-dependent K currents and slow tail currents of much slower time course. The rapidly decaying voltage-dependent tail currents were blocked with aminopyridines, and measurements of the slow tail currents were made following decay of any residual rapid tail currents. The slow tail current exhibited two components of differing sensitivity to externally applied tetraethylammonium (TEA) ions. In some neurones of the abdominal ganglion (L-2, L-4), virtually all of the slow tail current was resistant to blockage by TEA, while in others (L-3, L-6) 80% or more of the slow tail current was blocked by low TEA concentrations (KD less than 1 mM), the remaining slow tail current being resistant to TEA. This TEA-resistant slow tail current was identified as a K current because it reversed near the K equilibrium potential (EK), the reversal potential was shifted by changes in the external K concentration, and it could be blocked by injection of Cs+. It was abolished by replacement of external Ca2+ by Co2+ or Ba2+, by addition of Cd2+, or by injection of EGTA, and thus determined to be a Ca-dependent current. Intracellular injection of TEA or external application of aminopyridine or apamine had little or no effect on the TEA-resistant slow tail current. Quinidine reduced the TEA-sensitive, but not the TEA-resistant current. Both the TEA-sensitive and the TEA-resistant components of the slow tail current exhibited similar time courses of decay.(ABSTRACT TRUNCATED AT 250 WORDS)
Conjugates of the A-chain of the mistletoe lectin I and histamine but not A-chain per se inhibit the capacity of spleen cells of the mouse to induce antibody response or graft-versus-host reaction by 90%. The conjugates killed 27% of the spleen cells, 60% of the T-, and 15% of the B-lymphocytes. The receptors for histamine could be shown to be of the H2-type, because only H2-antagonists inhibited the toxic influence of the conjugates. The results suggest the hypothesis that not only suppressor cells but at least the majority of the immunocompetent cells bear receptors for histamine. The A-chain-histamine conjugate represents an "affinotoxin" which is cytotoxic only after affinity binding on receptors of the target cell.
Computer-assisted modeling of calcium influx through voltage-activated membrane channels predicted that buffer-limited elevation of cytoplasmic free calcium ion concentration occurs within microscopic hemispherical "domains" centered upon the active Ca channels. With increasing depolarization, the number of activated channels, and hence the number of Ca domains, should increase; the single-channel current should, however, decrease, thereby decreasing Ca2+ accumulation in each domain relative to the macroscopic current. Such voltage dependence of the microscopic distribution of Ca2+ may influence relations between total Ca2+ entry and Ca-dependent processes. Ca-mediated inactivation of Ca channels in Aplysia neurons exhibits behavior consistent with the calcium domain hypothesis.
The ability of Ca, Sr and Ba ions to support transmitter release was studied at the squid giant synapse by examining their respective actions on presynaptic current and post-synaptic responses. Transmitter-induced post-synaptic currents were smaller in Sr- than in Ca- containing solutions, and much smaller in Ba-containing solutions. The time course and amplitude of spontaneous miniature post-synaptic potentials were similar in the presence of all three divalent ions. Sr or Ba substitution has little effect on the resting potential of presynaptic terminals. In Sr-containing solutions, action potentials were similar in amplitude and time course to those recorded in Ca. Ba slightly prolonged action potential duration but had no effect on amplitude. Voltage-clamped presynaptic terminals exhibited inward Ca, Sr or Ba currents which were apparently carried through Ca channels. These currents were similar in amplitude and time course in all three ions, being somewhat larger in Ba. Although presynaptic currents were similar in these ions, transmitter release induced by these currents depended upon the divalent species entering the presynaptic terminal. Release was greatest in response to presynaptic current carried by Ca and smallest in response to current carried by Ba. Transfer curves relating presynaptic current to post-synaptic potential were sigmoidal in all three ions, and exhibited limiting slopes of approximately 2. Divalent cations differentially support transmitter release at the squid giant synapse in the sequence Ca greater than Sr much greater than Ba. The differential efficacy of the divalent cations is not due to post-synaptic alterations, presynaptic potential changes or differences in presynaptic divalent cation conductances. This sequence may reflect the cation selectivity of the exocytotic process responsible for transmitter release.
Ca currents flowing during voltage-clamp depolarizations were examined in axotomized Aplysia neurones under conditions that virtually eliminated other currents. Moderate to large currents exhibited a two-component time course of relaxation that can be approximated reasonably well by the sum of two exponentials. The rapid phase (tau 1 approximately equal to 70 ms at 0 mV) plus the slower phase (tau 2 approximately equal to 300 ms at 0 mV) ride upon a steady, non-inactivating current, I infinity. Conditions that diminish the peak current amplitude, such as reduced stimulus depolarization, inactivation remaining from a prior depolarization, or partial blockade of the Ca conductance by Cd, slowed both phases of inactivation, and all selectively eliminated the tau 1 phase, such that weak currents exhibited only the slower phase of decline. Injection of EGTA slowed both phases of inactivation, decreased the extent of the tau 1 phase, and increased the intensity of I infinity and of the current during the tau 2 phase. For a given voltage, the rate of inactivation increased as the peak current strength was increased, and decreased as the peak current strength was decreased. For a given peak current the rate of inactivation decreased as depolarization was increased. The relation of inactivation to prior Ca2+ entry was essentially linear for small currents, but decreased in slope with time during strong currents. The relation also became shallower with increasing depolarization, suggesting an apparent decrease in the efficacy of Ca in causing inactivation at more positive potentials. The basic kinetics of Ca current inactivation along with experimentally induced changes in those kinetics were simulated with a binding-site model in which inactivation develops during current flow as a function of the entry and accumulation of free Ca2+. This demonstrated that a single Ca-mediated process can account for the two-component time course of inactivation, and that the nearly bi-exponential shape need not arise from two separate processes. The two-component time course emerges as a consequence of a postulated hyperbolic reaction between diminishing probability of channels remaining open and the accumulation of intracellular free Ca2+. The occurrence of a single- or a two-component time course of inactivation thus appears to depend on the levels of internal free Ca2+ traversed during current flow.
Explore the source record for details and available documents.
The effect on the Ca-dependent potassium current, IK(Ca), of procedures that increase intracellular cAMP levels was studied in Aplysia neurons using three different pharmacological approaches. Exposure to cAMP analogues which were either resistant to or protected from phosphodiesterase hydrolysis caused an increase in IK(Ca) from 30 to 50% in 10 min. The degree of reversibility of this effect varied from complete with db cAMP to very little with pcpt cAMP. Exposure to cholera toxin, which stimulates the synthesis of endogenous cAMP, increased IK(Ca) 25% in 10 min and the effect was not reversible. Both approaches were effective in all seven neuron types studied. Application of serotonin plus phosphodiesterase inhibitor caused an increase in IK(Ca) in neuron R15 but not in the other neuron types. Application of pentylene tetrazole (PTZ) led to a decrease in IK(Ca). It is proposed that elevation of cyclic AMP mediates an increased sensitivity of the IK(Ca) channel to Ca ions.
Calcium tail currents were measured in axotomized Aplysia neurones L2-L6 using a two-electrode voltage clamp and micro-electrodes of specially low resistance. Measurements were made at -40 mV following depolarizing pulses of 7 or 10 ms duration in the presence of 45 microM-tetrodotoxin and 200 mM-tetraethylammonium. Symmetrical currents were eliminated by addition of digitally stored current traces produced in response to equivalent hyperpolarizations. The remaining current, identified as a tail current, was blocked by replacement of extracellular calcium with cobalt or manganese. Computer fits showed that the tail current closely approximated the sum of two exponentially decaying components. The first had a time constant, tau 1, of 0.38 +/- 0.05 ms, which may have been frequency-limited by the speed of the clamp; the second had a time constant, tau 2, of 2.0 +/- 0.8 ms. A more slowly decaying third tail current component (tau 3 = 30 ms), which developed more slowly, may be related to the non-specific current rather than the calcium current. The tau 1 and tau 2 components of the tail current lost amplitude with increasing pulse duration along an approximately bi-exponential time course that resembled the time course of relaxation of the calcium current during a prolonged depolarization. The slow third component of the tail current showed no such inactivation. The amplitudes of the first and second components of the calcium tail current both increased as sigmoidal functions of the test pulse voltage, reaching half maximum at +20 mV and plateauing above +60 mV. The voltage dependencies of the two components were similar. The rate of decay of the tau 1 component increased with increasing temperature and with increasing negative potential, whereas tau 2 showed little dependence on these parameters. The rates of decay of neither the tau 1 nor the tau 2 component were affected by large changes in the amplitude of the test depolarization or in the amplitude of the tail current or by injection of calcium ions or EGTA. Thus, the kinetics of the tail current as resolved under our conditions appear to be virtually independent of calcium-mediated inactivation. Activation time constants (tau m) predicted from tau 1 are 3 to 5 times longer than the values of tau m determined from the half-time to peak of activation. These kinetics are slower than those reported for Limnaea by factors of 2.5 to 3.5.(ABSTRACT TRUNCATED AT 400 WORDS)
Calcium tail currents, recorded at -40 mV after repolarization from 7 or 10 ms voltage-clamp depolarizations in axotomized Aplysia neurones in the presence of tetrodotoxin and tetraethylammonium, were used to investigate the inactivation of the calcium conductance without interference from contaminating potassium currents. Prior depolarization with a prepulse (V1) resulted in a reduction in size of the tail currents recorded following the test pulse (V2). The reduction occurred in both the fast (tau 1 less than 0.4 ms) and slow (tau 2 approximately equal to 2.0 ms) components of the tail current. The degree of inactivation remained constant when tail currents were measured at potentials ranging up to 30 mV on either side of the potassium equilibrium potential. Thus, any changes in potassium current must have contributed virtually nothing to the changes in tail current amplitude seen following presentation of the prepulse. Inactivation was greatest following prepulses to potentials (+10 to +40 mV) that produce maximal entry of calcium ions, and declined to about zero as the prepulse approached the calcium equilibrium potential. For V1 potentials above +50 mV, the prepulse caused an apparent short-term facilitation of V2 tail currents in EGTA-injected neurones. This effect, detected up to 50 ms following the pulse, is of uncertain origin. Pressure injection of calcium ions caused reduction in the size of the tail current, which was restored by subsequent injection of EGTA. Tail current amplitude was reduced by presentation of the prepulse for all test pulse voltages, but the measured inactivation declined exponentially towards a minimum with test pulses of increasingly positive potential. Removal of inactivation following a 200 ms prepulse was greatly accelerated by injection of EGTA. The EGTA-resistant inactivation remaining at short times decayed with a time constant of about 0.12 s. The relation of tail current reduction to prepulse voltage is consistent with the interpretation that the EGTA-resistant inactivation remaining at short times depends on entry of calcium ions during the prepulse, as does the EGTA-sensitive inactivation remaining at later times. It is proposed that the 'EGTA-resistant' phase of inactivation results from loading of EGTA with calcium ions near the inner surface of the membrane during sustained calcium entry, allowing the intracellular calcium concentration to rise. The results provide further evidence for a calcium-mediated inactivation of the calcium conductance.(ABSTRACT TRUNCATED AT 400 WORDS)
Beads of calf serum proteins (CSB) with antigen-antibody complexes or antigen by activation with glutaraldehyde were used as immunoadsorbents for the fractionation of mouse lymphocytes. T-lymphocytes could be separated by this method with high purity. The precursors of antibody-forming cells were completely eliminated from spleen and bone marrow cell suspensions. The unspecific binding of lymphocytes to CBS or Sepharose was similar and not selective for T or B lymphocytes.
Explore the source record for details and available documents.
The results reported here show the correlation between spleen (S) and body (B) weights in normal mice. This correlation is relevant to calculation of the spleen index in the graft-versus-host reaction (GVHR). By linear regression analysis a calibration curve B/S = f(B) could be calculated. This calibration curve provides a more reliable estimate of the strength of the GVHR since it compensated for the biological variance of the control values. This method of calculation further offers the possibility of pooling litters of different size and age (0-4 days postpartum) without loss of information due to additional variables.
Calcium ions enter and accumulate during depolarization of some cells, activating a potassium current, IK(Ca), that depends on the cytoplasmic concentration of calcium ions, [Ca]i. However, elevation of [Ca]i can depress IK(Ca) elicited by a subsequent membrane depolarization. The depression of IK(Ca) is ascribed here to a [Ca]i-mediated inactivation of the voltage-gated calcium conductance, which causes a net reduction in calcium ions available for the activation of IK(Ca). This suggests that other processes dependent on gated calcium entry may also be depressed by small background elevations in cytosolic free calcium ions.
Explore the source record for details and available documents.
Literature review. Mast cells and basophils are included not only in anaphylactic but also in complement activating and cell-mediated immune reactions. A histamine release can be induced via the IgE mechanism, the anaphylatoxins, the histamine releasing activity, and by other reactions. Histamine and several other mediators induce a local inflammation reaction either by a direct action on their target cells or indirectly by the activation of other humoral and/or cellular effector systems. Histamine influences several immune reactions. Here, its influence on histamine receptor-bearing lymphocytes is discussed more in detail. Using in vitro test systems, it has been shown that histamine has a dual effect, it induces a suppression when applied in higher concentrations, while low concentrations are stimulating. In vivo, histamine receptor-bearing lymphocytes have suppressive functions, mediated by H2 receptors. Some preliminary evidence for the inclusion on histamine in the immunoreagulation of patients with several diseases is presented.
This paper describes common used methods of immune cell separation such as separation by density-gradient, cell electrophoresis, cell adherence, affinity chromatography, isolation of cell populations by antisera and complement, by cell sorting, by monolayer and lectins. The authors give a critical evaluation in respect of yield and purity in relation to the extent of the experimental procedure and regard eventual artefacts.