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L B Cohen

Publications and source records attributed to L B Cohen.

At least 109 records · Page 6Linked to original sources

Changes in light scattering that accompany the action potential in squid giant axons: potential-dependent components.

1. To obtain information about structural events that occur in axons, changes in light scattering from squid giant axons were measured during action potentials and voltage-clamp steps.2. The scattering changes were measured at several scattering angles. Because the changes in scattering divided by the resting scattering were between 10(-6) and 10(-5), signal-averaging techniques were used to increase the signal-to-noise ratio.3. The scattering changes during the action potential were different at different angles. Two types were found, one at 10-30 degrees (forward angles) and the other at 60-120 degrees (right angles).4. At forward angles, there was a transient scattering decrease during the action potential. The time course of the change was similar to that of the action potential; this change was thought to be potential-dependent.5. At right angles, there was a transient scattering increase during the action potential followed later by a second, longer-lasting increase. Indirect evidence indicated that neither component could be totally potential-dependent.6. To further analyse these effects, scattering was measured during voltage-clamp steps. The changes seen during hyperpolarizing steps were presumed to be potential-dependent; again two different changes were found, one at forward angles and one at right angles.7. The potential-dependent change at right angles occurred with a time course that could be approximated by a single exponential with a time constant tau = 24 musec. The change at forward angles required two exponentials, tau(1) = 23 musec, tau(2) = 900 musec, to represent its time course.8. The size of both potential-dependent changes was proportional to the square of potential. The change at right angles, but not that at forward angles, was increased in size by the addition of butanol or octanol to the bathing solution.

Action Potentials↗

Changes in axon light scattering that accompany the action potential: current-dependent components.

1. When light scattering was measured during hyperpolarizing and depolarizing voltage-clamp steps, relatively large scattering changes were found during the depolarizing steps. These large changes were found to depend on the time integral of the ionic current and not on the changes in conductance or potential.2. The current-dependent changes were examined at several scattering angles, and three distinct time courses were found. At 30-120 degrees , the main change occurred after the current when steps of 2-5 msec duration were used. This change was called I-90 degrees . At 15-30 degrees , the change occurred with the same time course as the time integral of the current. This change was called I-25 degrees . At 5-15 degrees the scattering change occurred with a time course intermediate between that of I-90 degrees and I-25 degrees . This change was called I-10 degrees .3. In all experiments, outward potassium and outward sodium currents led to similar light scattering changes indicating that specific effects of the cation carrying the current across the membrane were not involved.4. The size of I-90 degrees was reduced by 29% when an isethionate artificial sea water was substituted for the normal chloride artificial sea water. This reduction equalled the reduction predicted for a transport number effect at the membrane-solution interface. The time course of I-90 degrees was similar to the predicted time course for a volume change in the periaxonal space, and such volume changes were tentatively identified as the origin of I-90 degrees .5. Because of difficulties in measuring the time course of I-25 degrees , it was not possible to distinguish between a water of hydration effect and a transport number effect as the cause of this change. Similarly, the origins of I-10 degrees were not identified. Only I-10 degrees was altered in size and time course when the external refractive index was increased with bovine albumin.6. When the scattering changes during the action potential were examined in light of the voltage-clamp experiments, we concluded that the forward-angle change was potential-dependent and that the long-lasting change at right angles probably represented a swelling of the periaxonal space resulting from the fact that chloride carried a significant fraction of the outward current during the action potential.

Action Potentials↗

Changes in light scattering associated with the action potential in crab nerves.

1. Changes in light scattering from stimulated crab leg nerves were measured in an effort to study changes in structure that occur during the action potential.2. The peak changes in scattering after individual stimuli were 10(-4)-10(-5) of the resting scattering; signal averaging techniques were used to increase the signal-to-noise ratio.3. During the compound action potential there was a transient increase in the 90 degrees scattering which reached a peak about 20 msec after the stimulus. This increase was probably not due to a change in axon volume. Its time course was somewhat slower than that of birefringence changes measured at the same time.4. In normal sea water the transient increase was followed by a longer lasting (persistent) decrease in scattering. This persistent decrease could be converted to an increase by reducing the tonicity of the sea water or by raising the external refractive index. This scattering change may represent a swelling of the axon resulting from the ionic exchanges that occur during the action potential.

Action Potentials↗

Analysis of the potential-dependent changes in optical retardation in the squid giant axon.

1. An analysis has been made of the change in optical retradation of the membrane elicited by the application of voltage-clamp pulses in squid giant axons.2. The retardation response consists of three separate voltage-dependent components. For freshly mounted axons, defined as being in state 1, hyperpolarizing pulses give a rapid increase in the light intensity measured with crossed polarizers which has been termed the fast phase. This is followed by a rather slow return towards the base line termed the rebound. On treatment of the axon with certain agents that include tetrodotoxin, high calcium and terbium, the rebound disappears and the fast phase slows down, increases in size, and has a new slow component added to it. This transition from state 1 to a second state, 2, appears to be irreversible.3. In state 1, the time constant of the fast phase is 20-40 musec at 13 degrees C; it has a very large negative temperature coefficient (Q(10) = Ca.(1/8)). The size of the retardation change is independent of temperature and varies as the square of the applied voltage, but the voltage-retardation curve is symmetrical about a point well beyond zero membrane potential, at an internal potential of around + 70 mV. In state 2, the time constant is about five times larger, and varies much less markedly with temperature; the apex of the voltage-retardation curve is shifted to + 200 mV.4. The rebound has a time constant of the order of 20 msec at 13 degrees C. A 10 degrees rise in temperature more than halves the time constant and roughly doubles the amplitude of the rebound. The voltage dependence of the rebound differed from that of the fast phase.5. The slow component of state 2 has a time constant of about 2 msec which does not change noticeably between 10 and 25 degrees C. The size of this component seems to be linearly dependent on the applied voltage, rather than obeying a square law.6. A tenfold increase in external calcium concentration had no discernible effect on the fast and slow phases, but reversibly reduced the amplitude of the rebound nearly to half.7. In experiments on perfused axons, the retardation response was not measurably altered by any of the modifications made to the composition of the perfusing fluid.8. There was some indication of the possible existence of a small current- or conductance-dependent component of the retardation response.9. These phenomena seem likely to originate either from molecular relaxation processes analogous with the Kerr effect, or from changes in membrane thickness under the influence of the pressure exerted by the electric field. However, the specific molecules involved in the retardation response cannot yet be identified.

Animals↗

Changes in axon birefringence during the action potential.

1. Observations have been made on the changes in optical retardation accompanying the passage of impulses along crab leg nerves and squid giant axons.2. The nerves were mounted on the stage of a polarizing microscope, at 45 degrees to the planes of polarization and analysis, brightly illuminated with white light. During the nerve impulse the intensity of the light passing the analyser decreased temporarily by 1 part in 10(3)-10(6). Signal-averaging techniques were used to obtain an acceptable ratio of signal to noise.3. The changes in light intensity recorded under these conditions were shown to arise almost entirely from alterations in retardation, with little or no interference from scattering, absorption, linear dichroism or optical rotation effects; the occurrence of stimulus and coupling artifacts was also ruled out.4. In the squid giant axon, the retardation change was shown to be located in a thin cylinder immediately surrounding the axoplasm, and to have a radially oriented optic axis.5. The time course of the decrease in optical retardation was very similar to that of the action potential recorded with an intracellular electrode, suggesting that the retardation closely followed the electrical potential across the membrane.

Action Potentials↗

Light scattering and birefringence changes during activity in the electric organ of electrophorus electricus.

1. In an attempt to obtain information about structural changes related to electrical activity in Electrophorus electroplates, we have determined the size and time course of the changes in light scattering and in bire-fringence that occur during and after the discharge of the electric organ.2. The changes in light intensity detected with a photomultiplier were never greater than 0.2% for a single discharge, and were often much smaller than this, but records with an acceptable ratio of signal to noise could be obtained by signal-averaging techniques.3. A single stimulus led to a decrease, then an increase, and finally another decrease in the light scattered by slices of the main electric organ. These three phases were designated E1, E2 and E3.4. E1 started at the beginning of the action potential, and its peak was reached at the same time as the completion of repolarization, even when the repolarization was delayed by cooling or hastened by drawing larger currents from the tissue.5. E2 was proportional to the integral of the current flowing through the slice of electric organ, and may arise from the swelling and shrinking of the tubules that stud the faces of the electroplates. It developed within a millisecond or two of the start of an applied current, and lasted for about 100 msec.6. E3 was a variable decrease in scattering that lasted for some seconds.7. A stimulus also led to a transient increase in the birefringence of the electric organ. The optical change followed the change in electrical potential across the innervated faces of the electroplates with a delay of somewhat under 50 musec.8. This voltage-dependent change in birefringence may arise from a Kerr effect (electric birefringence) in the membrane or from compression of the membrane.

Action Potentials↗