Sodium Movements in Perfused Squid Giant Axons : Passive fluxes

Sodium movements in internally perfused giant axons from the squid Dosidicus gigas were studied with varying internal sodium concentrations and with fluoride as the internal anion. It was found that as the internal concentration of sodium was increased from 2 to 200 mM the resting sodium efflux increased from 0.09 to 34.0 pmoles/cm²sec and the average resting sodium influx increased from 42.9 to 64.5 pmoles/cm²sec but this last change was not statistically significant. When perfusing with a mixture of 500 mM K glutamate and 100 mM Na glutamate the resting efflux was 10 ± 3 pmoles/cm²sec and 41 ± 10 pmoles/cm²sec for sodium influx. Increasing the internal sodium concentration also increased both the extra influx and the extra efflux of sodium due to impulse propagation. At any given internal sodium concentration the net extra influx was about 5 pmoles/cm²impulse. This finding supports the notion that the inward current generated in a propagated action potential can be completely accounted for by movements of sodium.     

Continuous conduction of impulses in peripheral myelinated nerve fibers

1. Conduction of impulses in peripheral myelinated fibers of a nerve trunk is a continuous process, since with uninjured nerve fibers: (a) within each internodal segment the conduction time increases continuously and linearly with increasing conduction distance; (b) the presence of nodes of Ranvier does not result in any detectable discontinuity in the conduction of the impulse; (c) the ascending phase of the spike always has an S shape and never presents signs of fractionation; (d) the shape and magnitude of the spike are constant at all points of each internodal segment. 2. Records have been presented of the external logitudinal current that flows during propagation of an impulse in undissected single nerve fiber (Fig. 6). 3. Propagation of impulses across a conduction block occurs with a readily demonstrable discontinuity.     

After-Potentials and Large Depolarizations of Single Nodes of Ranvier Treated with Veratridine

Potential differences between normal nodes of Ranvier (single fiber from the sciatic nerve of the frog, air-gap method) and a node exposed to 1 to 2.5 x 10^-6 gm veratridine per ml were measured. Negative after-potentials occurred immediately after application of the alkaloid when spike configuration and resting potential were virtually unchanged. The after-potentials decreased in magnitude and their time constant increased as the resting membrane was depolarized either by outward currents or by a train of impulses. Increase of (Na)_o markedly increased the amplitude of the after-potential. After prolonged application of veratridine or with higher concentrations, a large slow depolarization (rate of potential change about 7 mv per second) could be triggered by a train of impulses or even a single spike. This depolarization could promptly be terminated by withdrawing Na. It is concluded that, once the nodal membrane has become permeable to Na (as during a spike), veratridine prevents the normal return of P_Na to its resting value.     

Photosensitivity of the Frog Iris

The absorption of quanta by rhodopsin leads to the contraction of frog iris muscle. The contractions reach a maximum after about 8 sec. in the light. When the light is turned off the irises relax exponentially with a half-time of about 6 sec. Membrane polarization is not necessary for the response but calcium movement and membrane permeability changes probably are. The response is not mediated by acetylcholine or epinephrine. The curves of log It vs. log t for constant response amplitude bend progressively upward away from a unit slope line at short times as larger response criteria are used because (a) light influences tension development over longer times and (b) the higher intensity, shorter duration flashes are less effective.     

Further study of soma,dendrite, and axon excitation in single neurons

The present investigation continues a previous study in which the soma-dendrite system of sensory neurons was excited by stretch deformation of the peripheral dendrite portions. Recording was done with intracellular leads which were inserted into the cell soma while the neuron was activated orthodromically or antidromically. The analysis was also extended to axon conduction. Crayfish, Procambarus alleni (Faxon) and Orconectes virilis (Hagen), were used. 1. The size and time course of action potentials recorded from the soma-dendrite complex vary greatly with the level of the cell's membrane potential. The latter can be changed over a wide range by stretch deformation which sets up a "generator potential" in the distal portions of the dendrites. If a cell is at its resting unstretched equilibrium potential, antidromic stimulation through the axon causes an impulse which normally overshoots the resting potential and decays into an afternegativity of 15 to 20 msec. duration. The postspike negativity is not followed by an appreciable hyperpolarization (positive) phase. If the membrane potential is reduced to a new steady level a postspike positivity appears and increases linearly over a depolarization range of 12 to 20 mv. in various cells. At those levels the firing threshold of the cell for orthodromic discharges is generally reached. 2. The safety factor for conduction between axon and cell soma is reduced under three unrelated conditions, (a) During the recovery period (2 to 3 msec.) immediately following an impulse which has conducted fully over the cell soma, a second impulse may be delayed, may invade the soma partially, or may be blocked completely. (b) If progressive depolarization is produced by stretch, it leads to a reduction of impulse height and eventually to complete block of antidromic soma invasion, resembling cathodal block, (c) In some cells, when the normal membrane potential is within several millivolts of the relaxed resting state, an antidromic impulse may be blocked and may set up within the soma a local potential only. The local potential can sum with a second one or it may sum with potential changes set up in the dendrites, leading to complete invasion of the soma. Such antidromic invasion block can always be relieved by appropriate stretch which shifts the membrane potential out of the "blocking range" nearer to the soma firing level. During the afterpositivity of an impulse in a stretched cell the membrane potential may fall below or near the blocking range. During that period another impulse may be delayed or blocked. 3. Information regarding activity and conduction in dendrites has been obtained indirectly, mainly by analyzing the generator action under various conditions of stretch. The following conclusions have been reached: The large dendrite branches have similar properties to the cell body from which they arise and carry the same kind of impulses. In the finer distal filaments of even lightly depolarized dendrites, however, no axon type all-or-none conduction occurs since the generator potential persists to a varying degree during antidromic invasion of the cell. With the membrane potential at its resting level the dendrite terminals contribute to the prolonged impulse afternegativity of the soma. 4. Action potentials in impaled axons and in cell bodies have been compared. It is thought that normally the over-all duration of axon impulses is shorter. Local activity during reduction of the safety margin for conduction was studied. 5. An analysis was made of high frequency grouped discharges which occasionally arise in cells. They differ in many essential aspects from the regular discharges set up by the generator action. It is proposed that grouped discharges occur only when invasion of dendrites is not synchronous, due to a delay in excitation spread between soma and dendrites. Each impulse in a group is assumed to be caused by an impulse in at least one of the large dendrite branches. Depolarization of dendrites abolishes the grouped activity by facilitating invasion of the large dendrite branches.     

Sodium extrusion by internally dialyzed squid axons

A method has been developed which allows a length of electrically excitable squid axon to be internally dialyzed against a continuously flowing solution of defined composition. Tests showed that diffusional exchange of small molecules in the axoplasm surrounding the dialysis tube occurred with a half-time of 2–5 min, and that protein does not cross the wall of the dialysis tube. The composition of the dialysis medium was (mM): K isethionate 151, K aspartate 151, taurine 275, MgCI₂ 4–10, NaCl 80, KCN 2, EDTA 0.1, ATP 5–10, and phosphoarginine 0–10. The following measurements were made: resting Na influx 57 pmole/cm²sec (n = 8); resting potassium efflux 59 pmole/ cm²sec (n = 4); stimulated Na efflux 3.1 pmole/cm²imp (n = 9); stimulated K efflux 2.9 pmole/cm²imp (n = 3); resting Na efflux 48 pmole/cm²sec (n = 18); Q ₁₀ Na efflux 2.2 (n = 5). Removal of ATP and phosphoarginine from the dialysis medium (n = 4) or external application of strophanthidin (n = 1) reversibly reduced Na efflux to 10–13 pmole/cm²sec. A general conclusion from the study is that dialyzed squid axons have relatively normal passive permeability properties and that a substantial fraction of the Na efflux is under metabolic control although the Na extrusion mechanism may not be working perfectly.     

Hyperpolarization of a barnacle photoreceptor membrane following illumination

Membrane potential changes following illumination of a photoreceptor cell in the lateral ocellus of a barnacle (Balanus eburneus) were studied by means of intracellular recording and polarization techniques. Illumination produces a depolarizing response. When the illumination is terminated, the membrane potential temporarily becomes more negative than the resting potential prior to illumination. Although the amplitude of this postillumination hyperpolarization depends upon the intensity as well as the duration of the light pulse, the time course is fairly constant. The hyperpolarization is not associated with any significant membrane conductance increase and is abolished by 10^-5 M ouabain. It diminishes when the external Na or K ions are removed. An intracellular injection of Na ions produces a hyperpolarization similar to that following illumination. It is suggested that the postillumination hyperpolarization is produced by an electrogenic Na pump which is activated by the Na influx during illumination.     

Electrical signs of impulse conduction in spinal motoneurons

An analysis has been made of the electrical responses recorded on the surface and within the substance of the first sacral spinal segment when the contained motoneurons are excited by single and repeated antidromic ventral root volleys. A succession of negative deflections, designated in order of increasing latency m, i, b, d, has been found. Each of those deflections possesses some physiological property or properties to distinguish it from the remainder. Indicated by that fact is the conclusion that the successive deflections represent impulse conduction through successive parts of the motoneurons that differ in behavior, each from the others. Since the spinal cord constitutes a volume conductor the negative deflections are anteceded by a positive deflection at all points except that at which the axonal impulses first enter from the ventral root into the spinal cord. Frequently two or more negative deflections are recorded together in overlapping sequence, but for each deflection a region can be found in which the onset of that deflection marks the transition from prodromal positivity to negativity. Deflection m is characteristic of axonal spikes. Latent period is in keeping with known axonal conduction velocity. Refractory period is brief. The response represented by m is highly resistant to asphyxia. Maximal along the line of ventral root attachment and attenuating sharply therefrom, deflection m can be attributed only to axonal impulse conduction. Deflection i is encountered only within the cord, and is always associated with a deflection b. The i,b complex is recordable at loci immediately dorsal to regions from which m is recorded, and immediately ventral to points from which b is recorded in isolation from i. Except for its great sensitivity to asphyxia, deflection i has properties in common with those of m, but very different from those of b or d. To judge by properties i represents continuing axonal impulse conduction into a region, however, that is readily depolarized by asphyxia. Deflection b possesses a unique configuration in that the ascending limb is sloped progressively to the right indicating a sharp decrease in velocity of the antidromic impulses penetrating the b segment. A second antidromic volley will not conduct from i segment to b segment of the motoneurons unless separated from the first by nearly 1 msec. longer than is necessary for restimulation of axons. This value accords with somatic refractoriness determined by other means. Together with spatial considerations, the fact suggests that b represents antidromic invasion of cell bodies. Deflection d is ubiquitous, but in recordings from regions dorsal and lateral to the ventral horn, wherein an electrode is close to dendrites, but remote from other segments of motoneurons, d is the initial negative deflection. In latency d is variable to a degree that demands that it represent slow conduction through rather elongated structures. When associated with deflection b, deflection d may arise from the peak of b with the only notable discontinuity provided by the characteristically sloped rising phase of b. Deflection d records the occupation by antidromic impulses of the dendrites. Once dendrites have conducted a volley they will not again do so fully for some 120 msec. Embracing the several deflections, recorded impulse negativity in the motoneurons may endure for nearly 5 msec. When the axonal deflection m is recorded with minimal interference from somatic currents, it is followed by a reversal of sign to positivity that endures as long as impulse negativity can be traced elsewhere, demonstrating the existence of current flow from axons to somata as the latter are occupied by impulses. Note is taken of the fact that impulse conduction through motoneurons is followed by an interval, measurable to some 120 msec., during which after-currents flow. These currents denote the existence in parts of the intramedullary motoneurons of after-potentials the courses of which must differ in different parts of the neurons, otherwise nothing would be recorded. The location of sources and sinks is such as to indicate that a major fraction of the current flows between axons and somata. For approximately 45 msec. the direction of flow is from dendrites to axons. Thereafter, and for the remaining measurable duration, flow is from axons to dendrites.     

On the decay of lateral phase separations in biological membranes

A theory for the decay of a lateral phase separation in a biological membrane has been developed based upon the edge decay of a population of circular molecular domains of uniform size. The theory has been applied to the case of vesicle fusion at a presynaptic membrane. It is shown that the efficiency of fusion decays exponentially in time with a rate constant $\text{solπ}\text{2τ}\text{N}{}^{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ which decreases as the rate at which bonds are broken within each domain (τ^?1) decreases and as the number of molecules within each domain (N) increases. Moreover, it has been speculated that this mechanism may offer in part an explanation for the slow, exponential decay during post-tetanic potentiation where it is known that the efficiency of neurotransmitter release at the presynaptic membrane is rate-controlling and decays exponentially.     

Synaptic inhibition in an isolated nerve cell

Following the preceding studies on the mechanisms of excitation in stretch receptor cells of crayfish, this investigation analyzes inhibitory activity in the synapses formed by two neurons. The cell body of the receptor neuron is located in the periphery and sends dendrites into a fine muscle strand. The dendrites receive innervation through an accessory nerve fiber which has now been established to be inhibitory. There exists a direct peripheral inhibitory control mechanism which can modulate the activity of the stretch receptor. The receptor cell which can be studied in isolation was stimulated by stretch deformation of its dendrites or by antidromic excitation and the effect of inhibitory impulses on its activity was analyzed. Recording was done mainly with intracellular leads inserted into the cell body. 1. Stimulation of the relatively slowly conducting inhibitory nerve fiber either decreases the afferent discharge rate or stops impulses altogether in stretched receptor cells. The inhibitory action is confined to the dendrites and acts on the generator mechanism which is set up by stretch deformation. By restricting depolarization of the dendrites above a certain level, inhibition prevents the generator potential from attaining the "firing level" of the cell. 2. The same inhibitory impulse may set up a postsynaptic polarization or a depolarization, depending on the resting potential level of the cell. The membrane potential at which the inhibitory synaptic potential reverses its polarity, the equilibrium level, may vary in different preparations. The inhibitory potentials increase as the resting potential is displaced in any direction from the inhibitory equilibrium. 3. The inhibitory potentials usually rise to a peak in about 2 msec. and decay in about 30 msec. After repetitive inhibitory stimulation a delayed secondary polarization phase has frequently been seen, prolonging the inhibitory action. Repetitive inhibitory excitation may also be followed by a period of facilitation. Some examples of "direct" excitation by the depolarizing action of inhibitory impulses are described. 4. The interaction between antidromic and inhibitory impulses was studied. The results support previous conclusions (a) that during stretch the dendrites provide a persisting "drive" for the more central portions of the receptor cell, and (b) that antidromic all-or-none impulses do not penetrate into the distal portions of stretch-depolarized dendrites. The "after-potentials" of antidromic impulses are modified by inhibition. 5. Evidence is presented that inhibitory synaptic activity increases the conductance of the dendrites. This effect may occur in the absence of inhibitory potential changes.     

A new electrogenic step in the ubiquinol: Cytochrome c2 oxidoreductase complex of Rhodopseudomonas sphaeroides

Myxothiazol, an inhibitor of the ubiquinol oxidase site of the ubiquinol: cytochrome c₂ oxidoreductase complex, has been shown in the present work to inhibit a part of the electrogenic process indicated by phase III of the carotenoid change, in addition to the part of the change inhibited by antimycin. This finding shows that there is an antimycin-insensitive, but myxothiazol-sensitive portion of the slow phase, which indicates the existence of an electrogenic event within the ubiquinol: cytochrome c₂ oxidoreductase complex, in addition to that linked to oxidation of cytochrome b-561 which has been previously characterized. Redox titrations show that the appearance of the new electrogenic step is correlated with the amount of cytochrome b-561 available in the oxidized form before the flash. The rate of the antimycin-insensitive and myxothiazol-sensitive portion of the carotenoid change correlates well with the rate of reduction of cytochrome b-561. No carotenoid change associated with reduction of cytochrome b-566 was seen. These findings suggest that the newly identified electrogenic process is linked to electron transfer between cytochrome b-566 and b-561. Calculations of the contribution of this new electrogenic step to the total electrogenic event within the complex show that electrons passing from cytochrome b-566 to cytochrome b-561 pass about 35–50% of the distance across the whole membrane.     

Calcium exchange and contraction strength of guinea pig atrium in normal and hypertonic media

A Krebs-Henseleit (KH) medium made hypertonic by adding nonpermeant molecules substantially increased the isometric peak tension at steady-state contractions below 3 per sec in guinea pig atrium at 27°C. Action potential durations were decreased. KH plus 100 mM raffinose or sucrose resulted in similar and nearly maximal changes which were essentially reversible upon return to normal KH. When one active contracting atrium was used to passively stretch a second atrium, the difference in Ca ion exchange (1 min exchange with the extracellular space) between active and stretched atria significantly increased at 1 per sec and at 2 per sec in going from normal to 100 mM hypertonic KH. The calculated mean Ca ion cellular exchange per beat per 100 g of cells (a) doubled in changing from normal to 100 mM hypertonic KH, and (b) decreased slightly in changing from contractions of 1 per sec to 2 per sec in normal KH. These data are consistent with the hypothesis (a) that Ca ion entry per beat from the extracellular space is proportional to membrane depolarized time with a constant medium and a steady-state condition, and the hypothesis (b) that 100 mM hypertonicity doubles the Ca ion entry rate during depolarization. These data enable rejection of the hypothesis that the peak tension is proportional to the Ca ion entry per beat from the extracellular space under steady-state conditions, and suggest that any additional Ca ion involved in the larger contractions at higher frequencies comes from an increase in Ca ion available from intracellular stores.     

Cat Heart Muscle in Vitro : VIII. Active transport of sodium in papillary muscles

The cells of cat right ventricular papillary muscles were depleted of K and caused to accumulate Na and water by preincubation at 2–3°C. The time courses of changes in cellular ion content and volume and of the resting membrane potential (V_m) were then followed after abrupt rewarming to 27–28°C. At physiological external K concentration ([K]_o = 5.32 mM) recovery of cellular ion and water contents was complete within 30 minutes, the maximal observable rates of K uptake and Na extrusion (Δmmol cell ion/(kg dry weight) (min.)) being 3.4 and 3.6, respectively. The recovery rate was markedly slowed at [K]_o = 1.0 mM. Rewarming caused V_m measured in cells at the muscle surface to recover within from <1 to 9 minutes, but only slight restoration of cellular ion contents (measured in whole muscles) had occurred after 10 minutes. Studies of recovery in NaCl-free sucrose Ringer''s solution made it possible to separate the ouabain-insensitive outward diffusion of Na as a salt from a simultaneous ouabain-sensitive Na extrusion which is associated with a net cellular K uptake. A hypothesis consistent with these observations is that rewarming may activate a ouabain-sensitive "electrogenic" mechanism, most probably the net active transport of Na out of the cell, from which net K uptake may then follow passively.     

La conversion analogique numérique des récepteurs à l'étirement de la patte métathoracique du criquet Schistocerca gregaria

During the liarmonic stimulation of locust stretch-receptors of the metathoracic tibio-femoral joint, a decrease in the number of potentials in a sensory discharge can be observed with increasing frequency of stretching. On a log-log scale, a straight line of negative slope is calculated and fits best for each unit of both receptors (DPLR: dorso-postero-lateral receptor and DALR: dorso-antero-lateral receptor). These different graphs indicate that the number of impulses per cycle (N) decreases as a power function of the frequency of the sinusoidal input (F): logN = −alogF+b. For one unit, the slope a does not depend on the amplitude, the steady-state level (carrier rate: v), or temperature. The constant b gives a measure of the steady-state level at the start of stimulation, whereas a determines the dynamic sensitivity of the tested cell. When a unit is stimulated at the tonic frequency value of its steady-state level, only one spike is phase-locked to the stimulus, especially if the amplitude of the movement is large. Furthermore, when the frequency of stimulation is reduced to a half, the number of impulses per cycle doubles for the B and C units. With the method we have described here, one can hope to get further objective values to classify and to express the sensitivity of different kinds of mechanoreceptors.     

Interpretation of the Repetitive Firing of Nerve Cells

Eccentric cells of Limulus respond with repetitive firing to sustained depolarizing currents. Following stimulation with a step of current, latency is shorter than first interval and later intervals increase progressively. A shock of intensity twice threshold can evoke firing 25 msec. after an impulse. But in the same cell, a current step twice rheobase evokes a second impulse more than 50 msec. after the first, and current intensity must be raised to over five times rheobase to obtain a first interval of about 25 msec. Repetitive firing was evoked by means of trains of shocks. With stimuli of moderate intensity, firing was evoked by only some of the shocks and intervals between successive impulses increased with time. This is ascribed to accumulation of refractoriness with successive impulses. Higher frequencies of firing are obtained with shocks of intensity n x threshold than with constant currents of intensity n x rheobase. It is concluded that prolonged currents depress the processes leading to excitation and that (in the cells studied) repetitive firing is controlled both by the after-effects of firing (refractoriness) and by the depressant effects of sustained stimuli (accommodation). Development of subthreshold "graded activity" is an important process leading to excitation of eccentric cells, but is not the principal factor determining frequency of firing in response to constant currents.     

Maintained activity in the cat''s retina in light and darkness

Nervous activity has been recorded from the unopened eye of decerebrate cats. Recordings were made with metal electrodes or with small micropipettes from ganglion cells or nerve fibers. Continuous maintained discharges were seen in all ganglion cells during steady illumination of their receptive fields, as well as in complete darkness. Possible artefacts, such as electrode pressure, abnormal circulation, anesthetic, and several other factors have been excluded as the source of the maintained discharge. Visual stimuli are therefore transmitted by modulating the ever present background activity. Discharge frequencies were measured following changes of retinal illumination. No consistent patterns of frequency change were found. The maintained discharge frequency may be permanently increased or decreased, or may remain practically unchanged by altering the steady level of illumination. In addition, there were often transient frequency changes during the first 5 to 10 minutes after changing illumination, before a final steady rate was established. A statistical analysis of the impulse intervals of the maintained discharge showed: (a) the intervals were distributed according to the gamma distribution (Pearson's type III), (b) the first serial correlation coefficient of the intervals was between –0.10 and –0.24, with a mean value of –0.17, which is significantly different from zero, (c) the higher order serial correlation coefficients were not significantly different from zero. Thus the firing probability at any time depends on the times of occurrence of the two preceding impulses only, and in such a way as to indicate that each impulse is followed by a transient depression of excitability that outlasts the following impulse. The possible sites at which spontaneous or maintained activity may originate in the retina are discussed.     

Time and concentration dependence of the dicyclohexylcarbodiimide inhibition of proton movements in the cytochromebc 1 complex from yeast mitochondria reconstituted into proteoliposomes

The cytochromebc ₁ complex was isolated from yeast mitochondria solubilized with the detergent dodecyl maltoside and reconstituted into proteoliposomes to measure electrogenic proton pumping. Optimal respiratory control ratios of 4.0, obtained after addition of the uncoupler CCCP, and H⁺/e ^– ratios of 1.6 were obtained when the proteoliposomes were prepared with egg yolk phosphatidylcholine supplemented with cardiolipin. Moreover, it was critical to remove excess dodecyl maltoside in the final concentrated preparation prior to reconstitution to prevent loss of enzymatic activity. The rate of electrogenic proton pumping, the respiratory control ratios, and the H⁺/e ^– ratios were decreased by incubation of the cytochromebc ₁ complex with dicyclohexylcarbodiimide (DCCD) in a time and concentration dependent manner. Maximum inhibitions were observed when 50 nmol DCCD per nmol of cytochromeb were incubated for 30 min at 12°C with the intact cytochromebc ₁ complex. Under these same conditions maximum labeling of cytochromeb with [¹⁴C] DCCD was reported in a previous study [Beattieet al. (1984).J. Biol. Chem. 259, 10562–10532] consistent with a role for cytochromeb in electrogenic proton movements.     

Influence of Lithium Ions on the Transmembrane Potential and Cation Content of Cardiac Cells

The effect of lithium ions on cardiac cells was investigated by recording the changes in transmembrane potential and by following the movement of Li, Na, and K across the cell membrane. Isolated preparations of calf Purkinje fibers and cat ventricular muscles were used. Potentials were measured by intracellular microelectrodes; ion transport was estimated by flame photometric analysis and by using the radioactive isotopes of Na and K. It was shown (a) that Li ions can replace Na ions in the mechanism generating the cardiac action potential but that they also cause a marked depolarization and pronounced changes in action potential configuration; (b) that the resting permeability to Li ions is high and that these ions accumulate in the cell interior as if they were not actively pumped outwards. In Li-Tyrode [K]_i decreases markedly while the K permeability seems to be increased. In a kinetic study of net K and Na fluxes, the outward movement of each ion was found to be proportional to the second power of its intracellular concentration. The effect on the transmembrane potential is explained in terms of changes in ion movement and intracellular ion concentration.     

Early Picosecond Events in the Photocycle of Bacteriorhodopsin

The primary processes of the photochemical cycle of light-adapted bacteriorhodopsin (BR) were studied by various experimental techniques with a time resolution of 5 × 10^-13 s. The following results were obtained. (a) After optical excitation the first excited singlet state S₁ of bacteriorhodopsin is observed via its fluorescence and absorption properties. The population of the excited singlet state decays with a lifetime τ₁ of ~0.7 ps (430 ± 50 fs) (52). (b) With the same time constant the first ground-state intermediate J builds up. Its absorption spectrum is red-shifted relative to the spectrum of BR by ~30 nm. (c) The second photoproduct K, which appears with a time constant of τ₂ = 5 ps shows a red-shift of 20 nm, relative to the peak of BR. Its absorption remains constant for the observation time of 300 ps. (d) Upon suspending bacteriorhodopsin in D₂O and deuterating the retinal Schiff base at its nitrogen (lysine 216), the same photoproducts J and K are observed. The relaxation time constants τ₁ and τ₂ remain unchanged upon deuteration within the experimental accuracy of 20%.     

In vivo plant impedance measurements and characterization of membrane electrical properties: The influence of cold acclimation

A procedure to measure both the real (resistance) and complex (reactance) part of plant tissue impedance is described. Bare metal electrodes were used and the impedance of electrodes was determined by obtaining measurements at four interelectrode distances. The frequency (f) dependence of the impedance of birdsfoot trefoil (Lotus corniculatus L.) stems could be modeled, approximately, by a resistor representing an extracellular resistance, a resistor representing an intracellular resistance, and a resistor and capacitor representing cell membranes. However, the membrane parameters appeared to be frequency dependent. Therefore, they were characterized by calculating a time constant at each frequency. For example, for one plant stem the time constant (τ) decreased from 1.19 × 10⁻³ sec at 101 hz to 1.36 × 10⁻⁶ sec at 100 KHz. This decrease with frequency could be described by an equation of the form: ln(τ) = a + bln(f) + c(ln[f])². Cold acclimation increased (P ⩽ 0.05) the intracellular resistance. But cold acclimation did not have a significant effect on estimates of the extracellular resistance, the membrane resistance, or the membrane time constant. Depending on the cultivar, cold acclimation either decreased or increased the estimate of membrane capacitance.