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F Ramón

Publications and source records attributed to F Ramón.

36 records · Page 2Linked to original sources

Interaction of anaesthetics with electrical synapses.

Studies of the interaction of anaesthetics with various preparations, from whole animals to organic solvents, have been continuing since Overton and Meyer found a correlation between anaesthetic potency and solubility in olive oil. Although the physiological basis of anaesthesia is far from clear, one popular hypothesis is that anaesthetics act primarily by interfering with the normal functioning of chemical synapses. This hypothesis is supported by experiments showing that these synapses are more sensitive to both local and general anaesthetics than are axons. The effects of anaesthetics on electrical synapses (gap-junctions or nexus) have not previously been studied. These ubiquitous structures, presumably responsible for cell-to-cell communication, are found in most vertebrate and invertebrate tissues. We report here the effects of several anaesthetics on electronic coupling between nerve cells, and show that electrical synapses are less sensitive to most anaesthetics than are chemical synapses and axonal membranes.

Alcohols↗

On the electrotonic coupling mechanism of crayfish segmented axons: temperature dependence of junctional conductance.

It is generally accepted that the mechanism for electrotonic coupling involves the presence of hydrophilic channels connecting the cytoplasm of neighboring cells. These channels are presumed to be water filled holes. To test this hypothesis, we measured the temperature dependence of coupling parameters and calculated the specific resistance of junctional synapses of crayfish segmented axons. Results demonstrate that: (i) low temperature increases the junctional resistance in a manner that depends on the time course of cooling; (ii) the specific junctional resistance is, at most, 1-20 omega cm2. These results are consistent with a hypothesis of cell communication based on hydrophilic channels and suggest the presence of a temperature-dependent component of these channels.

Action Potentials↗

[L.D.H. and isoenzymes in C.S.F. of children with neurological diseases (author's transl)].

Authors studied L.D.H .and Isoenzymes of L.D.H. activities in 254 C.S.F. corresponding to 112 children affected with different diseases. The purpose was to discover its diagnostic utility and possible etiological explanation. Results have been compared with reference values found in a previous study. Highest enzymatic activities of L.D.H. are observed, in decreasing order, in tumours, terminal process, endocranial hipertension plus meningitis, and meningitis. Referring to different types of meningitis, highest enzymatic activity of L.D.H. was found in bacterial type, especially those caused by a germen different from "Neisseria meningitidis". An inversion of isoenzymatic pattern is observed with an increase of L.D.H.-4 and L.D.H.-5 in detriment of L.D.S.-1 and L.D.S.-2, in tumours, endocranial hipertension plus meningitis and in all infections of central nervous system in the initial phase, returning to normal in recuperation phase of disease. A possible enzymatic etiology in different pathological diseases is suggested. In conclusion, it is of interest to determine L.D.H. and its isoenzymes as control of disease but not as differential diagnosis.

Child↗

Propagation of action potentials in squid giant axons. Repetitive firing at regions of membrane inhomogeneities.

Effects of reduction in potassium conductance on impulse conduction were studied in squid giant axons. Internal perfusion of axons with tetraethylammonium (TEA) ions reduces G K and causes the duration of action potential to be increased up to 300 ms. This prolongation of action potentials does not change their conduction velocity. The shape of these propagating action potentials is similar to membrane action potentials in TEA. Axons with regions of differing membrane potassium conductances are obtained by perfusing the axon trunk and one of its two main branches with TEA after the second branch has been filled with normal perfusing solution. Although the latter is initially free of TEA, this ion diffuses in slowly. Up until a large amount of TEA has diffused into the second branch, action potentials in the two branches have very different durations. During this period, membrane regions with prolonged action potentials are a source of depolarizing current for the other, and repetitive activity may be initiated at transitional regions. After a single stimulus in either axon region, interactions between action potentials of different durations usually led to rebound, or a short burst, of action potentials. Complex interactions between two axon regions whose action potentials have different durations resembles electric activity recorded during some cardiac arrhythmias.

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[Reference values of A.S.T., L.D.H., isoenzymes of L.D.H. and A.D.A. in cerebrospinal fluid of children (author's transl)].

From 230 C.S.F., authors found the reference values of A.S.T., L.D.H., isoenzymes of L.D.H. and A.D.A. in children from the area of Barcelona. The reference values obtained were: A.S.T.: n, 208; mean, 6.91 U/l., and s, +/- 2.71 U/l. A.D.A.: n, 175; percentil 50, 0.388 U/l.; percentil 97.5, 1.64 U/l., and percentil 2.5, 0 U/l. L.D.H.: n, 148; percentil 50, 9.54 U/l.; percentil 97.5, 24.96 U/l., and percentil 2.5, 3.81 U/l. Isoenzymes of L.D.H.: n, 82; L.D.H.-1: mean, 41.31 per cent and s, +/- 5.41 per cent. L.D.H.-2: mean, 30.79 per cent, and s, +/- 2.84 per cent. L.D.H.-3: mean, 20.3 per cent, and s, +/- 4.11 per cent. L.D.H.-4: mean, 6.02 per cent, and s, +/- 2.48 per cent. L.D.H.-5: mean, 2.47 per cent, and s, +/- 1.7 per cent. Higher activities of A.S.T. and L.D.H. were found in newborns, also varying the distribution of isoenzymes of L.D.H. The A.D.A didn't show variations.

Adenosine Deaminase↗

Fine structure of the electrotonic synapse of the lateral giant axons in a crayfish (Procambarus clarkii).

The electrotonic synapse of lateral giant axons of the crayfish was studied by conventional thin sectioning. The most prominent membrane specialization observed in this synaptic region is the communicating junction. It is characterized by a close apposition of the two axonal membranes which are separated by a 4--5 nm wide gap. Other characteristics of the junction are an array of particles spaced about 20--22 nm apart and a row of vesicles symmetrically arranged at the cytoplasmic leaflets of each membrane. The communicating junction does not cover the entire surface of the electrotonic synapse. Indeed, we have found other specializations such as: finger-like Schwann cell processes extending between synaptic membranes, saccular invaginations of one synaptic membrane into its axon, and coated vesicles continuous with one of the membranes. In addition, large vesicular pieces of the communicating junctions, with their accompanying vesicles, appeared to extend deeply inside the axoplasm. The morphological appearance of the communicating junction is found to be different from the one reported for mammalian maculae communicans such as liver or heart muscle. This is surprising because, regardless of their morphological differences, both junctions seem to transmit electrotonically.

Animals↗

Ephaptic transmission in squid giant axons.

Some characteristics of ephaptic transmission of action potentials were investigated with squid giant axons. For these studies two isolated axons were placed side by side or, on occasion, a single long axon was looped to form an "ephapse" between the axon trunk and one of its main branches. Extracellular potentials measured adjacent to axons surrounded by a very restricted volume of liquid ranged up to 80 mV in magnitude and had a shape similar to that of the membrane current. Intracellular records of the same axon regions show small voltage deflections; however, the transmembrane voltage (Vm = Vi - Vo) has the appearance of normally propagated action potentials. Ephaptic transmission of action potentials is possible when the ephaptic region is submerged in oil, as well as when the region is immersed in low-calcium solutions. When the speed of the propagated action potential is lowered by replacing the normal artifical seawater (ASW) with low-sodium ASW, some ephaptic effects are enhanced. It is concluded that in regions in which axons are confined by restricted extracellular volume, the large extracellular voltage changes arising during the passage of an action potential in one can cause ephaptic excitation in another.

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Squid giant axons. A model for the neuron soma?

Insertion of electrically floating wires along the axis of a squid giant axon produces an apparent increase in diameter in the region where the wire surface has been treated to give it a low resistance. The shape of action potentials propagating into this region depend upon the surface resistance (and the length) of the wire. As this segment's internal resistance is lowered by reducing the wire's surface resistance, the following characteristic sequence of changes in the action potential is seen at the transition region: (a) the duration increases; (b) two peaks develop, the first one generated in the normal axon region and the second one generated later in the axial wire region, and; (c) blockage occurs (for a very low resistance wire). Action potentials recorded at the membrane region near the tip of the axial wire in (b) resemble those recorded at the initial segment of neurons upon antidromic invasions. Squid axon action potentials propagated from a normal region into that containing the low resistance wire also resemble antidromic invasions recorded in neuron somas. Hyperpolarizing current pulses applied through the wire act as if the wire surface resistance was momentarily reduced. For example, the two components of the action potential recorded at the axial wire membrane region noted in (b) can be sequentially blocked by the application of increasing hyperpolarizing current through the wire. Similar effects are seen when hyperpolarizing currents are injected into motoneuron somas. It is concluded that the geometrical properties of the junction of a neuron axon with its soma may be in themselves sufficient to determine the shape of the action potentials usually recorded by microelectrodes.

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Axon voltage-clamp simulations. I. Methods and tests.

This is the first in a series of four papers in which we present the numerical simulation of the application of the voltage clamp technique to excitable cells. In this paper we describe the application of the Crank-Nicolson (1947) method for the solution of the parabolic partial differential equations that describe a cylindrical cell in which the ionic conductances are functions of voltage and time (Hodgkin and Huxley, 1952). This method is compared with other methods in terms of accuracy and speed of solution for a propagated action potential. In addition, differential equations representing a simple voltage-clamp electronic circuit are presented. Using the voltage clamp circuit equations, we simulate the voltage clamp of a single isopotential membrane patch and show how the parameters of the circuit affect the transient response of the patch to a step change in the control potential. The stimulation methods presented in this series of papers allow the evaluation of voltage clamp control of an excitable cell or a syncytium of excitable cells. To the extent that membrane parameters and geometrical factors can be determined, the methods presented here provide solutions for the voltage profile as a function of time.

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Axon voltage-clamp simulations. II. Double sucrose-gap method.

This is the second in a series of four papers on the simulation of the voltage clamp of cylindrical excitable cells. In this paper we evaluate the double sucrose-gap voltage-clamp technique for the squid and lobster giant axons. Using the Crank-Nicolson method of solution of the cable equations and differential equations representing the voltage clamp circuit we studied the effect of length of the sucrose gap "node" on the voltage profile along an excitable cell during a simulated voltage clamp. The voltage gradients along the region of the cell within the node produce "notches" in the current recording as well as changes in the magnitude of the sodium and potassium current for a given voltage step. Our results show that good voltage clamp control requires node lengths less than one-half the axon diameter.

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Axon voltage-clamp simulations. III. Postsynaptic region.

This is the third in a series of four papers in which we present the numerical simulations of the application of the voltage clamp technique to excitable cells. In this paper we discuss the problem of voltage clamping a region of a cylindrical cell using microelectrodes for current injection and voltage recording. A recently developed technique (Llinás et al., 1974) of internal application of oil drops to electrically insulate a short length of the postsynaptic region of the squid giant synapse is evaluated by simulation of the voltage clamp of an excitable cylindrical cell of finite length with variable placement of the current and voltage electrodes. Our results show that ENa can be determined quite accurately with feasible oil gap lengths but that the determination of the reversal potential for the synaptic conductance, ES, can be considerably in error. The error in the determination of ES dependp, and especially the membrane resistance at the time the synaptic conductance occurs. It is shown that the application of tetraethylammonium chloride to block the active potassium conductance very significantly reduces the error in the determination of ES. In addition we discuss the effects of cable length and electrode position on the apparent amplitude and time course of the syn aptic conductance change. These results are particularly relevant to the application of the voltage clamp technique to cells with nonsomatic synapses. The method of simulation presented here provides a tool for evaluation of voltage clamp analysis of synaptic transmission for any cell with known membrane parameters and geometry.

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Axon voltage-clamp simulations. A multicellular preparation.

In this paper we extend the simulation of the voltage clamp of a single nerve fiber to a bundle of axons. These simulations included not only the description of the voltage clamp circuit and a single unidimensional cable to represent the preparation in the "node" region of a double sucrose gap used previously but also a series resistance and a shunt pathway. The output of the voltage control amplifier is applied across the membrane plus the series resistance, producing a voltage drop across the series resistance due to the current generated by the membrane in response to a depolarizing voltage step. Since the membrane current has an inward and an outward phase, voltage drops of opposite sign are produced across the series resistance. During the transient current and at all points along an axon, the potential deviation produced by the series resistance is opposite to the deviation produced by the longitudinal gradient. Only at a command potential equal to the sodium equilibrium potential, the membrane potential transiently matches the command potential. For the attempted voltage clamp of an axon, values of series resistance larger than 50 omega-cm2 allowed propagated action potentials in the membrane. In spite of the presence of propagated action potentials at the calbe membrane, the recorded current does not show "notches" and it has a phase of inward current and a phase of outward current. It is concluded that, in a multicellular preparation with series resistance, the recording of a square voltage pulse does not indicate voltage control of the transmembrane potential. The presence of a shunt pathway produces inaccurate values of current density. Neither series or shunt resistance produce "notches" in the current records.

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On the connection between the transverse tubules and the plasma membrane in frog semitendinosus skeletal muscle. Are caveolae the mouths of the transverse tubule system?

The transverse tubular system (TTS) of skeletal muscle fibers represents the morphological basis for the inward spread of conduction of the electrical signal that triggers muscle contraction. A historical account of the main steps contributing to the elucidation of the structure and function of the TSS has been presented by Huxley (1971). While the localization of the TSS and its association with the sarcoplasmic reticulum (SR) is well documented; there is still a need further to develop our knowledge of the morphology of the connection between the TSS and the plasma membrane. It is generally believed that the TSS opens directly to the extracellular space and that there is continuity between its membrane and the sarcolemma. However, direct observation of such a connection has been clearly shown only for the myotome of fish (Franzini-Armstrong and Porter, 1964). In other muscle fibers, only indirect evidence of the connection has been provided by experiments showing penetration of extracellular tracers into the TSS. These extracellular markers were also observed inside another membrane-bounded compartment consisting of round profiles named "caveolae" (Yamada, 1955) or "pinocytotic vesicles" (Ashurst, 1969). The present study deals with the communication between the TTS, caveolae, and plasma membrane (Peachey, 1965); Ezerman and Ishikawa, 1967; Schiaffino and Margreth, 1968; and Rayns et al., 1968). A detailed study of the caveolae compartment was undertaken with ruthenium red as an electron-dense tracer. As a result of this study, we propose that in certain species the caveolae compartment represents the transitional region in the connection between the TSS and the sarcolemma.

Animals↗

Simulation of action potential propagation in an inhomogeneous sheet of coupled excitable cells.

Cable theory and active equivalent circuits have been used to simulate the propagation of action potentials along a single nerve or muscle fiber by representing the cell as a unidimensional cable composed of isopotential segments. We extended this method to a two-dimensional sheet of cells which in many ways represents the atrium. Our method consisted of solving for the potential profile of a sheet composed of a large number of isopotential membrane patches, each of which was represented by an active equivalent circuit in which the ionic conductances were functions of voltage and time. The patches were arranged in a rectangular array with resistive interconnections that could be varied over the sheet. We used this model to study the effect of various inhomogeneities on conduction velocity and the resulting wave fronts in a sheet of excitable tissue. Some of these inhomogeneities included different effective internal resistances in the x and y directions, preferential pathways, and discrete regions of changing resistive connections. The results showed that very localized changes in membrane properties or cellular interconnections produce changes in the wave front over broad areas. This model provides a method for computing the wave fronts of action potential propagation in any two-dimensional inhomogeneous sheet of coupled excitable cells.

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Propagation of action potentials in inhomogeneous axon regions.

Described are studies of propagation of action potentials through inhomogenous axon regions through experiments performed on squid giant axons and by computer simulations. The initial speed of propagation of the action potential is dependent upon the stimulus waveform. For a rectangular pulse of current, the action potential travel initally at a high speed that declines over the distance, reaching a constant speed of propagation at about 1-5 resting length constants; this distance depends on the stimulus strength. additional experiments studied the effects of changing the axon diameter and of introducing a temperature step. It was found that the propagated action potential suffers profound modification in shape and velocity as it reaches the region of transition. In both cases, it was possible to obtain reflected action potentials. A region of increased effective diameter was produced experimentally in the squid giant axon by insertion of an axial wire as usually employed in voltage clamps. It was found that the action potential, at the axial wire tip region, undergoes shape changes similar to those obtained tn simulations of a region of increased diameter as in a junction with the axon and soma in motor neurons. It is conducluded that the gaint axon can be used to reproduce simple electrical behaviors in other structures.-Ramón, F., R. W. Joyner and J.W. Moore. Propagation of action potentials in inhomogeneous axon regions.

Action Potentials↗