Photocurrent kinetics of purple-membrane sheets bound to planar bilayer membranes

Summary The kinetics of light-driven proton transport by bacteriorhodopsin has been studied in a model system consisting of a planar lipid bilayer membrane to which purple membrane fragments have been attached. After excitation with a 10-nsec laser flash a fast negative current-transient occurs, followed by a positive transient which decays to zero. The time course of the photocurrent may be represented by a sum of four exponentials with time constants ₁= 1.2sec, ₂= 17sec, ₄= 57sec, ₁= 950sec (at 25°C). In a D₂O medium ₂ and ₃ are increased by a factor of 2.6 and 2.9, respectively, whereas ₁ remains unaffected. The observed components of the photocurrent can be correlated to photochemical reaction steps inferred from flash-photometric experiments on the basis of the observed time constants, the activation energies, and the effects of pH and D₂O. From the photocurrent signals information may be obtained on the magnitude of the charge displacement associated with the elementary transitions of the bacteriorhodopsin molecule.     

Modelling endplate currents: dependence on quantum secretion probability and decay of miniature current

Quantification of the time course and amplitude of endplate currents (EPC) was made with respect to dispersion of quanta secretion and to changes in the exponential decay of miniature endplate currents (_mepc). The relationship between RPC amplitude and _mepc follows a double-exponential curve with ₁= 0.3 ms and ₂ = 6 ms. If the amplitude of fully synchronised EPC is taken as 100%, then the loss of EPC amplitude is already 42% with physiological parameters of dispersion (the half-rise and decay constant of distribution of secretion probability = 0.5 ms, _mepc =1 ms). This loss is even more substantial if secretion is more dispersed or miniature endplate currents decay faster. Correspondence to: F. Vyskocil     

Conformational dynamics of the anticodon loop in yeast tRNAPhe as sensed by the fluorescence of wybutine

Conformational and dynamic properties of the anticodon loop of yeast tRNA^Phe were investigated by analyzing the time resolved fluorescence of wybutine serving as a local structural probe adjacent to the anticodon GmAA on its 3 side. The influence of Mg²⁺, important for stabilizing the tertiary structure of tRNA, and of the complementary anticodon s²UUC of E. coli tRNA ₂ ^Glu were investigated.Fluorescence lifetimes and anisotropies were measured with ps time resolution using time correlated single photon counting and a mode locked synchronously pumped and frequency doubled dye laser as excitation source. From the analysis of lifetimes () and rotational relaxation times ( _R ) we conclude that wybutine occurs in various structural states: (i) one stacked conformation where the base has no free mobility and the only rotational motion reflects the mobility of the whole tRNA molecule (=6 ns, _R =19 ns), (ii) an unstacked conformation where the base can freely rotate (=100 ps, _R = 370 ps) and (iii) an intermediary state (=2 ns, _R = 1.6 ns).Under biological conditions, i. e. in the presence of Mg²⁺ and neutral salts, wybutine is found in a stacked and immobile state which is consistent with the crystallographic picture. In the presence of the complementary codon however, as exemplified by the E. coli-tRNA ₂ ^Glu anticodon, our analysis indicates that the codon-anticodon complex exists in an equilibrium of structural states with different rotational mobility of wybutine. The conformation with wybutine freely mobile is the predominant one and suggests that this conformation of the codon-anticodon structure differs from the canonical 3–5 stack.     

Comparison of 15N- and 13C-determined parameters of mobility in melittin

Backbone and tryptophan side-chain mobilities in the 26-residue, cytolytic peptide melittin (MLT) were investigated by ¹⁵N and ¹³C NMR. Specifically, inverse-detected ¹⁵N T₁ and steady-state NOE measurements were made at 30 and 51 MHz on MLT at 22 °C enriched with ¹⁵N at six amide positions and in the Trp¹⁹ side chain. Both the disordered MLT monomer (1.2 mM peptide at pH 3.6 in neat water) and -helical MLT tetramer (4.0 mM peptide at pH 5.2 in 150 mM phosphate buffer) were examined. The relaxation data were analyzed in terms of the Lipari and Szabo model-free formalism with three parameters: _m, the correlation time for the overall rotation; S², a site-specific order parameter which is a measure of the amplitude of the internal motion; and _e, a local, effective correlation time of the internal motion. A comparison was made of motional parameters from the ¹⁵N measurements and from ¹³C measurements on MLT, the latter having been made here and previously [Kemple et al. (1997) Biochemistry, 36, 1678–1688]. _m and _e values were consistent from data on the two nuclei. In the MLT monomer, S² values for the backbone N-H and C-H vectors in the same residue were similar in value but in the tetramer the N-H order parameters were about 0.2 units larger than the C-H order parameters. The Trp side-chain N-H and C-H order parameters, and _e values were generally similar in both the monomer and tetramer. Implications of these results regarding the dynamics of MLT are examined.     

Effects of moisture stress on the multiplication and expansion of cells in leaves of sugar beet

Summary Sugar beets were subjected to moisture stress by decreasing the water potential of the culture solution osmotically with polyethylene glycol by a known amount, , and, alternatively by applying matric potential, , at the plant roots. Lowering the water potential at the root surface less than 200 millibars by either method resulted in significant decreases in the rate of cell multiplication. The final number of cells per leaf at = -372 mb the final was 165% of that at = -473 mb ( = –101 mb); similarly at = –15 mb the final cell number was 198% of that at = –196 mb ( = –181 mb). The mean cell volume of leaves was not significantly affected by these levels of moisture stress.     

Application of the Criterion of Biological Time (τ n /τ0) to the Studies of Embryogenesis in Birds

It was shown on the example of chick embryo that the number ₀ ( _n /₀) can be recommended as a measure of biological time and, for this purpose, the duration of the minimal mitotic cycle during synchronous cleavage divisions should be determined (in minutes) in various avian species. Based on the preliminary data, one can propose the comparability and similarity of the temporal programs of gastrulation and somitogenesis in the chick embryo and embryos of some fish and amphibians.     

"Velocity leakage" in the pigeon vestibulo-ocular reflex

The transfer characteristics of the vestibuloocular reflex (VOR), and of the semicircular canal primary afferents (SCPAs) that drive it, have been studied in several species. In monkeys and cats, the dominant time constant describing horizontal VOR dynamics ( _hu ) is longer than that ( _c ) of horizontal SCPAs. This lengthening of the time constant has been attributed to a velocity storage mechanism that has been modeled as a positive feedback loop in the VOR pathways. We have studied the transfer characteristics of horizontal and vertical VOR and SCPAs in unanesthetized pigeons. In this species the dominant time constants of both the horizontal and vertical VOR ( _hv and _vv ) are shorter than _c . This finding indicates that time constants characterizing the lower frequency response of the VOR can be lengthened or shortened depending on the species. We propose that in the pigeon the velocity leakage mechanism can be modeled by substituting negative feedback for positive feedback in the model of the VOR pathways. Negative feedback can also account for the further shortening of _hu and _vv as VOR gain increases with arousal. Additionally, making the negative feedback loop nonlinear can model the dependency of lower frequency VOR phase on amplitude, and skew in VOR waveforms. Pigeon VOR and SCPA dynamics also differ in their adaptive properties and higher frequency behavior. A predominance of input from highly adaptive SCPAs is proposed to account for the increased adaptation of the vertical VOR as compared with SCPAs overall. A pure time-delay associated with VOR operation can explain the phase lag of the VOR relative to SCPAs at higher frequencies.     

Die Aktivit?tsperiodik bei V?geln mit und ohne dunklem Schlafkasten

Zusammenfassung In 2 Versuchsserien wurden Kohlmeisen(Parus major) und japanische Möwchen(Lonchura striata var.domestica) einzeln und schallisoliert gehalten. In der ersten Versuchsserie, in der alle Vögel einen dunklen Schlafkasten hatten, wurde der Einfluß der Beleuchtungsstärke auf die Periode () der Hüpfaktivität und auf das Verhältnis von Aktivitätszeit zu Ruhezeit ( : -Verhältnis) untersucht. Sowhol Kohlmeisen als auch japanische Möwchen folgen der Regel, daß mit wachsender Beleuchtungsstärke die Periode kürzer und das : -Verhältnis größer wird.In der 2. Serie wurde der Einfluß Ruhe im dunklen Schlafkasten auf die Periodenlänge und auf das : -Verhältnis untersucht. Es wurden die Messungen aus Bedingung 1 (der Vogel hat einen dunklen Schlafkasten zur Verfügung) mit den Messungen aus Bedingung 2 (der Vogel hat keinen oder einen hellen Schlafkasten zur Verfügung) verglichen. Das Ergebnis bei Kohlmeisen entspricht den Befunden bei konstantem Licht verschiedener Intensität. Unter Bedingung 1 ist länger und das : -Verhältnis kleiner als in Bedingung 2. Das Ausmaß der Änderung von nach Fortnahme des dunklen Kastens ist unabhängig von der Periodenlänge in Bedingung 1. Das Ausmaß der änderung von : ist unabhängig von a : in Bedingung 1, jedoch schwach negativ korreliert mit der Periodenlänge in Bedingung 1.Bei japanischen Möwchen entsprechen die Ergebnisse dieser Versuchsserie nicht der Regel für tagaktive Vögel. Mit Benützen des dunklen Schlafkastens ist kürzer als ohne den Schlafkasten. Ohne den Schlafkasten ist etwa 24 Std. Das : -Verhältnis ist in Bedingung 1 unter bestimmten Voraussetzungen kleiner als in Bedingung 2. Das Ausmaß der Änderung von nach Fortnahme des Kastens ist mit der dazugehörigen Periode in Bedingung 1 hochsignifikant korreliert (Regressionskoeffizient b=-1.01, Korrelationskoeffizient r=0.89). Ebenfalls ist das Ausmaß der Änderung von : nach Fortnahme des Kastens mit : aus Bedingung 1 korreliert; es scheint, als würde ein bevorzugtes : -Verhältnis von etwa 2.0 eingeregelt.Die Ergebnisse werden im Hinblick auf 4 Punkte diskutiert: 1) Das circadiane System arbeitet innerhalb eines engen Bereiches von - und : -Werten optimal. 2) Der Optimalbereich wird bevorzugt unter ungünstigen Bedingungen angestrebt. 3) Der Entzug des dunklen Schlafkastens belastet japanische Möwchen mehr als Kohlmeisen. 4) Bei japanischen Möwchen wird in Bedingung 1 durch fortplfanzungsphysiologischen Einfluß verkürzt.

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Circadian activity rhythms of birds with and without a dark nest box

Summary Perch-hopping activity of Great tits(Parus major) and Bengales finches(Lonchura striata domestica), housed individually in soundproof boxes, was studied in two series of experiments. In the first series all birds had access to a dark nest box, in which they retired during their subjective night. In this experiment the effect of light intensity on the freerunning circadian activity rhythm was investigated. Both Great tits and Bengalese finches obey the circadian rule by responding to an increase in light intensity with shortening the circadian period () and with an increase of the ratio of activity time and rest time ( : ).In the second series of experiments the influence of sleeping in the dark nest box on both circadian period and : -ratio was studied. The results of two experimental conditions — without and with access to a dark nest box — were compared. In the Great tits, the results are in agreement with the effect of light intensity: when a dark nestbox is available, is longer and the : -ratio is smaller than in the absence of a nest box. The magnitude of the change in free-running period after removal of the nest box is independent of the original value of ; the amount of change : -ratio is likewise independent of the original : -ratio, but is weakly correlated to the original .InLonchura striata var. domestica, removal of the dark nest box leads to a lenghtening of the free-running period to about 24 hours; the : -ratio is smaller in the presence of a dark nestbox, if certain other conditions are fulfilled. The magnitude of the change in after removal of the nest box is highly correlated to the original free-running period (r=-0.89) in such a way that, without nest box, the period approaches a value of 24 hours. Also, the amount of change in : -ratio due to nest box removal is negatively correlated to the original : -ratio. A probably preferred : -ratio of 2.0 is adopted.These results are discussed in the view of 4 points: 1) The circadian system operates at its optimum within a narrow range of - and : -values. 2) This optimal range is especially adopted when conditions become adverse. 3) Removal of the dark nest box results in a more stressful situation for Bengalese finches than for Great tits. 4) In the Bengalese finches, is shortened in the presence of a nest box due to effects on reproductive physiology.

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Herrn Prof. Dr. JürgenAschoff zum 60. Geburtstag gewidemt.     

Residual motion of hemoglobin-bound spin labels and protein dynamics: viscosity dependence of the rotational correlation times

The residual motion of spin labels bound to cysteine 93 and to lysines of methemoglobin has been studied by electron paramagnetic resonance spectroscopy. To separate the influences of the solvent and the protein environment of the label fluctuations, the correlation times, , were analyzed as a function of temperature for fixed solvent viscosities, . Results show that over a wide range of viscosity the dependence of on may be empirically described by a power law ^k . The exponent k depends strongly on the location of the label on the protein surface. If one regards the spin labels as artificial amino acid side chains, characteristic values of correlation times and amplitudes of the rotational motion at the surface can be given. For =1 cP and T=297 K the correlation time of the labels bound to lysines is found to be =9 · 10^–10 s and the rotational diffusion is nearly isotropic. The spin label bound to cysteine 93 occupies a protein pocket, its rotational motion is therefore restricted. The correlation time of the label motion within a limited motion cone of semi angle =30° ± 3° is found to be =1.3 · 10^–9 s for =1 cP and T=297 K.     

Growth and reproduction of the Nile perch,Lates niloticus,an introduced predator,in the Nyanza Gulf,Lake Victoria,East Africa

Synopsis Length-frequency data suggest Nile perch, Lates niloticus, from the Nyanza Gulf grew to a total length of 9 cm by age 118 days and 23 cm by age 287 days. A modified von Bertalanffy growth curve _t = 1.35·L(1-e^–K(t-t _o)) with the parameters L = 93.1, K = 0.272 and t_o = 0.046, is suggested to describe growth up to 5 years of age and the relationship _t = 1.35·(31.96 + 7.681t) for fish aged 6 years and above. Length-weight relationships were = 0.0234·-gt^2.74 for fish between 7 and 15.9 cm total length, = 0.0151·^2.94 for fish between 16 and 45.9 cm total length, and = 0.0023·^3.44 for fish between 46 and 120 cm total length. Male Nile perch first matured between 50 and 55 cm total length when they were probably 2 years old; female Nile perch first matured between 80 and 85 cm total length when they were probably 4 years old. Small males were common, large males were rare, with the reverse holding for females. Sex change, from male to female, is a possible explanation for this size dimorphism.     

Existence and uniqueness of a sharp travelling wave in degenerate non-linear diffusion Fisher-KPP equations

In this paper we use a dynamical systems approach to prove the existence of a unique critical value c ^* of the speed c for which the degenerate density-dependent diffusion equation u _ct = [D(u)u _x ] _x + g(u) has: 1. no travelling wave solutions for 0 < c < c ^*, 2. a travelling wave solution u(x, t) = (x - c ^* t) of sharp type satisfying (– ) = 1, () = 0 ^*; '(^*–) = – c ^*/D'(0), '(^*+) = 0 and 3. a continuum of travelling wave solutions of monotone decreasing front type for each c > c ^*. These fronts satisfy the boundary conditions (– ) = 1, '(– ) = (+ ) = '(+ ) = 0. We illustrate our analytical results with some numerical solutions.     

Evaluation of errors of interproton distances and correlation time determined from NMR cross-relaxation rates

Summary We have analyzed a combined use of the two-dimensional nuclear Overhauser effect in the laboratory frame (NOESY) and in the rotating frame (ROESY) to determine interproton distances and correlation time in medium-sized rigid molecules (Davis, 1987). This method can be applied in the intermediate motional regime, 0.2 < _oc, < 5, (_c, correlation time, (_o resonance frequency). Error limits depend on the motional regime and are smallest near _oc=1.14.The method was tested on six geminal proton pairs in the bicyclic octapeptide (S-deoxo--[R]-OH-Ile³ amaninamide, Mw =870) for which at 297 K in DMSO, a correlation time of 1.0 ns, with a standard deviation of 0.12 ns, and an interproton distance of 1.87 Å, with standard deviation of 0.04 Å, are obtained.     

The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update

Mitochondrial dysfunction has been shown to participate in the induction of apoptosis and has even been suggested to be central to the apoptotic pathway. Indeed, opening of the mitochondrial permeability transition pore has been demonstrated to induce depolarization of the transmembrane potential (_m), release of apoptogenic factors and loss of oxidative phosphorylation. In some apoptotic systems, loss of _m may be an early event in the apoptotic process. However, there are emerging data suggesting that, depending on the model of apoptosis, the loss of _m may not be an early requirement for apoptosis, but on the contrary may be a consequence of the apoptotic-signaling pathway. Furthermore, to add to these conflicting data, loss of _m has been demonstrated to not be required for cytochrome c release, whereas release of apoptosis inducing factor AIF is dependent upon disruption of _m early in the apoptotic pathway. Together, the existing literature suggests that depending on the cell system under investigation and the apoptotic stimuli used, dissipation of _m may or may not be an early event in the apoptotic pathway. Discrepancies in this area of apoptosis research may be attributed to the fluorochromes used to detect _m. Differential degrees of sensitivity of these fluorochromes exist, and there are also important factors that contribute to their ability to accurately discriminate changes in _m.     

Induced diploid gynogenesis and polyploidy in the ornamental (koi) carp Cyprinus carpio L.

The results of a series of experiments conducted in our laboratory on the ornamental common carp (koi), aimed at optimizing heat-shock chromosome-set manipulation procedures, are described. The timing of heat-shock initiation was expressed in the relative unit of embryological age (₀) in order to standardize this parameter, the absolute time for heat-shock initiation being calculated from duration of one ₀ at two different pre-treatment water temperatures. Heat shocks were applied within the periods of 0.05–0.60 ₀ and 1.20–2.20 ₀ which, respectively, cover the successive phases of the 2nd meiotic division and the 1st cleavage. The highest production of diploid gynogenetic offspring was observed when heat shocks were initiated at 0.15–0.25 ₀ and at 1.5 ₀, after insemination, corresponding to anaphase of meiosis-II, and metaphase of the 1st cleavage, respectively. Similar results were obtained irrespective of the different pre-treatment water temperatures, thus confirming the possibility of standardizing heat-shock timing by ₀.     

Effects of zwitterionic detergents on the electronic structure of the primary donor and the charge recombination kinetics of P+Q A − in native and mutant reaction centers from Rhodobacter sphaeroides

The effects of detergents on the electronic structure of the oxidized primary donor P⁺ and the time constant _AP of the P⁺Q _A ^– charge recombination at ambient temperatures have been investigated in native and mutant reaction centers (RCs) from Rhodobacter sphaeroides. It is shown that N-lauryl-N,N-dimethyl-3-ammonio-1-propane sulfonate (SB12) induces a transition to a second distinct conformation of the RC. In the case of the wild type and the mutant FY(M197), in which a hydrogen bond is introduced to the 2-acetyl group of the dimer half of P that is associated with the M-subunit of the RC, the conformational change causes a more asymmetric spin density distribution between the two bacteriochlorophyll moieties of P⁺ in favor of the L-half. For both types of RCs the time constant _AP depends on the SB12/RC ratio as does the position of the long-wavelength band of P, _max. The increase of _AP by 30 ms and the shift of _max from 866 nm to 851 nm are indicative for the conformational change. In addition, a smaller linear increase of _AP with increasing SB12/RC ratio is superimposed on the variation of _AP due to the conformational change. Similar effects of SB12 on the optical spectra as well as on _AP are also observed for the two heterodimer mutants HL(L173) and HL(M202), in which one of the bacteriochlorophylls of P is replaced by a bacteriopheophytin. There are no clear indications for a correlation of _AP with the localization of the positive charge in P⁺. Furthermore, it is concluded from the dependence of _AP on the SB12/RC ratio that the single-site mutations do not affect the standard free energy difference of the two conformations to a measurable extent.     

A factor-dependent sulfotransferase specific for 3′-phosphoadenosine-5′-phosphosulfate (PAPS) in the Cyanobacterium Synechococcus 6301

A sulfotransferase isolated from the Cyanobacterium Synechococcus 6301 was found to be specific for 3-phosphoadenosine-5-phosphosulfate (PAPS). The molecular weight of this transferase has been estimated on a Sephadex-G-100 column to be about 58,000. The K _m for PAPS was determined to be 20 M. The pH optimum was 8.0. The thiol dithioerythritol was needed for activity; other thiols such as glutathione, cysteine, or mercaptoethanol did not catalyze this reaction. The transferase, however, could not react directly with the thiol. A heat-stable factor was needed in this reaction. This factor was purified by conventional techniques and its molecular weight was determined on a Sephadex-G-50 column to be about 11,500. The factor showed normal Michaelis-Menten behavior toward the PAPS-sulfotransferase. It has been identified as thioredoxin. The tranferase was inhibited by 3-5-ADP and 2–5-ADP; all other adenine-containing nucleotides such as 2-AMP, 3-AMP, 5-AMP, ADP, and c-AMP did not influence this reaction.Abbreviation PAPS 3-phosphoadenosine-5-phosphosulfate     

Comparison of Pre- and postsynaptic process kinetics in neuro-muscular synapse using the end-plate current model

The effect of correlation between the kinetics of acetylcholine (ACh) quanta release and the kinetics of the postsynaptic membrane channel activity on end-plate currents (EPCs) was studied in model experiments. The presynaptic process was described by a transmitter secretion probability distribution (SPD), and the postsynaptic process by characteristicsof miniature EPC (MEPC) with standard amplitude and variable decay duration (_MEPC). The SPD was represented by a curve with an S-like rise and exponential decay. The main portion of the data was analyzed after being adjusted to 20°C. The effect of the EPC amplitude loss (the difference between the EPC amplitudes in cases of synchronous and non-synchronous release of the ACh quanta) due to temporal dispersion of the transmitter secretion process was 43% at _MEPC=1.0 msec and 20% at _MEPC=4–5 msec under conditions when acetylcholinesterase (AChE) was inhibited.When _MEPC varied over a wide range, the effect of the EPC amplitude loss could be described by a curve with steeply and gently sloping portions; the boundary between these two portions corresponded to approximately 1.0 msec _MEPC, which is nearly the channel lifetime at resting membrane potential. The rise time of the EPC only slightly related to changes in _MEPC. The rate of EPC decay exceeded that in MEPC only when the lifetimes of the ACh-activated ionic channels were lower than their physiological values. In this case, the kinetics of transmitter release became a factor determining the EPC decay time course.Three variants of an increase in temporal dispersion of the ACh quanta secretion were modeled for constant _MEPC: one resulting from an increase in the SPD rising phase; a second resulting from equal increases in the SPD rising and declining phases, and a third resulting from an increase in the SPD declining phase. The results of the first variant corresponded most closely to experimental data on changes in EPC during a short-term period of rhythmical activity. This was also characterized by minimal effect of EPC amplitude loss.It has been suggested that the interrelation between the parameters of SPD and _MEPC is an important factor regulating the amplitude and duration of the postsynaptic signal. The maintenance of this interrelation at a constant level ensures reliability of synaptic transmission.Translated from Neirofiziologiya, Vol. 25, No. 2, pp. 126–132, March–April, 1993.     

Tunneling Processes Induced by Terahertz Electric Fields

Tunneling processes induced by terahertz frequency electric fields havebeen investigated.A drastic enhancement of the tunneling probabilityhas been observed by increasing the frequency at_e 1 where_e is the tunneling time.For a given constant tunneling rate an increase offrequency by a factor of seven leads to a drop of the requiredelectric field strengthby three orders of magnitude.It is shown that the enhancement of tunneling ionization at terahertz frequencies is due to the factthat electrons can absorb energy from the radiation field during tunnelingreducing the effective width of the tunneling barrier.     

Electrochemical potential of protons in vesicles reconstituted from purified,proton-translocating adenosine triphosphatase

Summary Measurements were made of the difference in the electrochemical potential of protons ( ) across the membrane of vesicles reconstituted from the ATPase complex (TF ₀ ·F ₁) purified from a thermophilic bacterium and P-lipids. Two fluorescent dyes, anilinonaphthalene sulfonate (ANS) and 9-aminoacridine (9AA) were used as probes for measuring the membrane potential () and pH difference across the membrane ( pH), respectively.In the presence of Tris buffer the maximal and no pH were produced, while in the presence of the permeant anion NO ₃ ^– the maximal pH and a low were produced by the addition of ATP. When the ATP concentration was 0.24mm, the was 140–150 mV (positive inside) in Tris buffer, and the pH was 2.9–3.5 units (acidic inside) in the presence of NO ₃ ^– . Addition of a saturating amount of ATP produced somewhat larger and pH values, and the attained was about 310 mV.By trapping pH indicators in the vesicles during their reconstitution it was found that the pH inside the vesicles was pH 4–5 during ATP hydrolysis.The effects of energy transfer inhibitors, uncouplers, ionophores, and permeant anions on these vesicles were studied.     

Effect of axoplasmic transport blockade on end-plate currents in frog muscle fibers

Evoked and spontaneous end-plate currents (EPC) were studied in normal voltage-clamped frog sartorius muscle fibers and 2 weeks after application of colchicine to the nerve innervating the muscle to block axoplasmic transport in its fibers. Application of colchicine was found to reduce the rate of rise and to prolong decay of EPC without affecting the amplitude of the EPC and miniature EPC, the quantum composition of EPC, and the frequency of miniature EPC. The histogram of distribution of the time constant () of EPC decay under normal conditions follows the normal law, but after application of colchicine to the nerve it is shifted to the right, with separation of two modes (₁ and ₂). Three types of synapses can be distinguished from the character of EPC decay: monoexponential decay with ₁ (44%), biexponential decay with ₁ and ₂ (39%), and monoexponential decay with ₂ (19%). An increase in of EPC decay is accompanied by strengthening of the dependence of this process on the clamping voltage. The current-voltage characteristic and reversal potential of EPC are unchanged. It is suggested that the change in character of EPC decay after application of colchicine to the motor nerve is due to the appearance of acetylcholine-activated ionic channels in the muscle membrane with a longer duration of the open state and with potential-dependence of the open state similar to that taking place after muscle denervation.S. V. Kurashov Medical Institute, Ministry of Health of the RSFSR, Kazan'. Translated from Neirofiziologiya, Vol. 17, No. 2, pp. 204–211, March–April, 1985.