scholarly journals Peroxidase uptake by photoreceptor terminals of the skate retina.

1976 ◽  
Vol 70 (1) ◽  
pp. 86-96 ◽  
Author(s):  
H Ripps ◽  
M Shakib ◽  
E D MacDonald
Keyword(s):  
Transmitter Release ◽  
Synaptic Vesicles ◽  
Light Adaptation ◽  
Ringer Solution ◽  
Bathing Solution ◽  
Synaptic Terminal ◽  
Vesicle Membrane ◽  
Electrical Potentials ◽  
Membrane Bound ◽  
Peroxidase Uptake

The photoreceptors of dark-adapted skate retinas bathed in a Ringer solution containing horseradish peroxidase (HRP) incorporate the tracer into membrane-bound compartments within the synaptic terminal of the cell; after 1 or 2 h of incubation, approx. 10-38% of the synaptic vesicles were labeled. The receptors appeared to be functioning normally throughout the incubation period, since electrical potentials of normal amplitude could be elicited in response to dimphotic stimuli. However, it was possible to block the uptake of peroxidase by a regimen of light adaptation that effectively suppressed light-induced activity in the electroretinogram. If, during incubation with peroxidase, retinas were exposed at 10-min intervals to an intense 1-ms flash from a xenon discharge tube, the receptor terminals were almost completely devoid of peroxidase; fewer than 2% of the vesicles were labeled. The suppression of HRP uptake could also be achieved in dark-adapted retinas by adding magnesium to the bathing solution, suggesting that calcium is necessary for transmitter release from vesicles in the receptor terminals. These findings are consistent with the view that vertebrate photoreceptors discharge a neurotransmitter in darkness, and that light decreases the release of this substance. It seems likely that the incorporation of peroxidase into vesicles of physiologically active receptor terminals reflects a mechanism for the retrieval of vesicle membrane after exocytosis.

scholarly journals Artificially imposed electrical potentials drive l-glutamate uptake into synaptic vesicles of bovine cerebral cortex

1990 ◽  
Vol 267 (1) ◽  
pp. 63-68 ◽  
Author(s):  
J Shioi ◽  
T Ueda
Keyword(s):  
Cerebral Cortex ◽  
Synaptic Vesicles ◽  
Glutamate Uptake ◽  
Electrical Potential ◽  
Potential Gradient ◽  
Protonmotive Force ◽  
Uptake System ◽  
Vesicle Membrane ◽  
Excitatory Neurotransmitter ◽  
Electrical Potentials

L-Glutamate is a major excitatory neurotransmitter in the central nervous system. MgATP-dependent glutamate uptake and H(+)-pumping ATPase activity were reported in highly purified synaptic vesicles [Naito & Ueda (1983) J. Biol. Chem. 258, 696-699; Shioi, Naito & Ueda (1989) Biochem. J. 258, 499-504], and it is hypothesized that an electrochemical H+ gradient across the vesicle membrane, the so-called protonmotive force, elicits the neurotransmitter uptake. An inside-positive diffusion potential across the vesicle membrane was established with valinomycin plus Rb+. This artificial electrical potential promoted the uptake of glutamate, but not aspartate, in the synaptic vesicles prepared from bovine cerebral cortex. The uptake was inhibited by the protonmotive-force dissipators carbonyl cyanide p-trifluoro-methoxyphenylhydrazone or nigericin, and was enhanced by concomitant imposition of a pH jump (alkalinization) in the external medium. Subcellular and subvesicular distributions showed the uptake system to be predominantly associated with small synaptic vesicles. The results support the hypothesis that glutamate uptake into synaptic vesicles is coupled with a H+ efflux down the electrochemical potential gradient, which is generated by H(+)-pumping ATPase.

scholarly journals QUANTITATIVE ASPECTS OF TRANSMITTER RELEASE

1970 ◽  
Vol 47 (3) ◽  
pp. 585-592 ◽  
Author(s):  
George D. Bittner ◽  
Donald Kennedy
Keyword(s):  
Motor Neuron ◽  
Transmitter Release ◽  
Synaptic Vesicles ◽  
Inhibitory Neuron ◽  
Release Mechanism ◽  
Quantal Content ◽  
Vesicle Membrane ◽  
Release Process ◽  
Membrane Materials ◽  
Order Of Magnitude

The opener-stretcher motor neuron in crayfish makes 50 endings upon each of 1200 muscle fibers. We have calculated the quantal content of junctional potentials produced by individual terminals and by the whole cell at various physiological frequencies. The results show that when the motor neuron is active at 20 impulses/second, it releases 50 quanta/impulse per muscle fiber, or a total of 4.5 x 109 quanta/hr. These figures are similar to those for vertebrate muscles per fiber, but larger for the entire neuron because the opener motor unit is so large. On the basis that the quanta correspond to synaptic vesicles each containing 103–104 molecules of transmitter, the release rate must be around 10-11 mole/hr. This value is within an order of magnitude of the release figures obtained for mammalian neurons by collecting transmitter in perfusates, but it is far lower than the value reported for a crustacean inhibitory neuron. If the membrane materials surrounding each vesicle were lost in the release process, the replacement synthesis would involve 24 mm2 of membrane/hr. We conclude that the metabolic load in terms of transmitter synthesis is probably sustainable, but that the release mechanism must operate in such a way that vesicle membrane materials are neither lost nor incorporated into the terminal membrane.

scholarly journals A marginal band of microtubules transports and organizes mitochondria in retinal bipolar synaptic terminals

2015 ◽  
Vol 146 (1) ◽  
pp. 109-117 ◽  
Author(s):  
Malkolm Graffe ◽  
David Zenisek ◽  
Justin W. Taraska
Keyword(s):  
Electron Microscopy ◽  
Transmitter Release ◽  
Synaptic Vesicles ◽  
Bipolar Cells ◽  
Synaptic Activity ◽  
Synaptic Terminal ◽  
Synaptic Terminals ◽  
Marginal Band ◽  
Microtubule Structure

A set of bipolar cells in the retina of goldfish contains giant synaptic terminals that can be over 10 µm in diameter. Hundreds of thousands of synaptic vesicles fill these terminals and engage in continuous rounds of exocytosis. How the cytoskeleton and other organelles in these neurons are organized to control synaptic activity is unknown. Here, we used 3-D fluorescence and 3-D electron microscopy to visualize the complex subcellular architecture of these terminals. We discovered a thick band of microtubules that emerged from the axon to loop around the terminal periphery throughout the presynaptic space. This previously unknown microtubule structure associated with a substantial population of mitochondria in the synaptic terminal. Drugs that inhibit microtubule-based kinesin motors led to accumulation of mitochondria in the axon. We conclude that this prominent microtubule band is crucial to the transport and localization of mitochondria into the presynaptic space to provide the sustained energy necessary for continuous transmitter release in these giant synaptic terminals.

Real-time measurement of transmitter release from single synaptic vesicles

Nature ◽  
1995 ◽  
Vol 377 (6544) ◽  
pp. 62-65 ◽  
Author(s):  
Dieter Bruns ◽  
Reinhard Jahn
Keyword(s):  
Real Time ◽  
Transmitter Release ◽  
Synaptic Vesicles ◽  
Time Measurement ◽  
Real Time Measurement

scholarly journals Changes in miniature endplate potential frequency during repetitive nerve stimulation in the presence of Ca2+, Ba2+, and Sr2+ at the frog neuromuscular junction.

1981 ◽  
Vol 77 (5) ◽  
pp. 503-529 ◽  
Author(s):  
J E Zengel ◽  
K L Magleby
Keyword(s):  
Nerve Stimulation ◽  
Transmitter Release ◽  
Neuromuscular Junctions ◽  
Fourth Power ◽  
Bathing Solution ◽  
Frog Neuromuscular Junction ◽  
Repetitive Nerve Stimulation ◽  
Time Constants ◽  
Endplate Potential ◽  
Induced Changes

Miniature endplate potentials (MEPPs) were recorded from frog sartorious neuromuscular junctions under conditions of reduced quantal contents to study the effect of repetitive nerve stimulation on asynchronous (tonic) quantal transmitter release. MEPP frequency increased during repetitive stimulation and then decayed back to the control level after the conditioning trains. The decay of the increased MEPP frequency after 100-to 200-impulse conditioning trains can be described by four components that decayed exponentially with time constants of about 50 ms, 500 ms, 7 s, and 80 s. These time constants are similar to those for the decay of stimulation-induced changes in synchronous (phasic) transmitter release, as measured by endplate potential (EPP) amplitudes, corresponding, respectively, to the first and second components of facilitation, augmentation, and potentiation. The addition of small amounts of Ca2+ or Ba2+ to the Ca2+-containing bathing solution, or the replacement of Ca2+ with Sr2+, led to a greater increase in the stimulation-induced increases in MEPP frequency. The Sr-induced increase in MEPP frequency was associated with an increase in the second component of facilitation of MEPP frequency; the Ba-induced increase with an increase in augmentation. These effects of Sr2+ and Ba2+ on stimulation-induced changes in MEPP frequency are similar to the effects of these ions on stimulation-induced changes in EPP amplitude. These ionic similarities and the similar kinetics of decay suggest that stimulation induced changes in MEPP frequency and EPP amplitude have some similar underlying mechanisms. Calculations are presented which show that a fourth power residual calcium model for stimulation-induced changes in transmitter release cannot readily account for the observation that stimulation-induced changes in MEPP frequency and EPP amplitude have similar time-courses.

Effect of Hypertonicity on Augmentation and Potentiation and on Corresponding Quantal Parameters of Transmitter Release

1999 ◽  
Vol 81 (3) ◽  
pp. 1428-1431 ◽  
Author(s):  
Hong Cheng ◽  
Michael D. Miyamoto
Keyword(s):  
Nerve Stimulation ◽  
Transmitter Release ◽  
Ringer Solution ◽  
Hypertonic Solution ◽  
Repetitive Nerve Stimulation ◽  
Quantal Transmitter Release ◽  
Before And After ◽  
Miniature Endplate Potentials ◽  
Endplate Potentials ◽  
Made In

Effect of hypertonicity on augmentation and potentiation and on corresponding quantal parameters of transmitter release. Augmentation and (posttetanic) potentiation are two of the four components comprising the enhanced release of transmitter following repetitive nerve stimulation. To examine the quantal basis of these components under isotonic and hypertonic conditions, we recorded miniature endplate potentials (MEPPs) from isolated frog ( Rana pipiens) cutaneous pectoris muscles, before and after repetitive nerve stimulation (40 s at 80 Hz). Continuous recordings were made in low Ca2+ high Mg2+ isotonic Ringer solution, in Ringer that was made hypertonic with 100 mM sucrose, and in wash solution. Estimates were obtained of m (no. of quanta released), n (no. of functional release sites), p (mean probability of release), and vars p (spatial variance in p), using a method that employed MEPP counts. Hypertonicity abolished augmentation without affecting potentiation. There were prolonged poststimulation increases in m, n,and p and a marked but transient increase in vars p in the hypertonic solution. All effects were completely reversed with wash. The time constants of decay for potentiation and for vars p were virtually identical. The results are consistent with the notion that augmentation is caused by Ca2+ influx through voltage-gated calcium channels and that potentiation is due to Na+-induced Ca2+ release from mitochondria. The results also demonstrate the utility of this approach for analyzing the dynamics of quantal transmitter release.

Structural and functional changes of frog neuromuscular junctions in high calcium solutions

1971 ◽  
Vol 178 (1053) ◽  
pp. 407-415 ◽  
Keyword(s):  
Transmitter Release ◽  
Synaptic Vesicles ◽  
Neuromuscular Junctions ◽  
Motor Nerve ◽  
Secretory Response ◽  
Spontaneous Release ◽  
Functional Changes ◽  
High Calcium ◽  
Release Of Acetylcholine ◽  
Evoked Transmitter Release

When frog muscles are exposed for several hours to a solution of isotonic calcium chloride, the secretory response of the motor nerve terminals to imposed depolarization ultimately fails and the rate of spontaneous release of acetylcholine also declines towards zero. The failure of depolarization-evoked transmitter release is irreversible while spontaneous release reappears, though in highly abnormal fashion, when the muscle is returned to a normal ionic medium. Examination of motor end-plates, during various stages of calcium treatment, shows that there is gradual intra-axonal agglutination of synaptic vesicles which is only very incompletely reversible. This effect is presumably the consequence of gradual entry and intracellular accumulation of calcium ions. Analogous treatment with isotonic magnesium, while resulting in immediate loss of evoked transmitter release, does not lead to progressive agglutination of synaptic vesicles, nor to irreversible impairment of the secretory response of the nerve terminal. The possible relations between structural and functional changes during calcium and magnesium treatment are discussed.

Tonic transmitter release in a graded potential synapse

1995 ◽  
Vol 74 (1) ◽  
pp. 470-473 ◽  
Author(s):  
R. O. Uusitalo ◽  
M. Juusola ◽  
E. Kouvalainen ◽  
M. Weckstrom
Keyword(s):  
Noise Reduction ◽  
Transmitter Release ◽  
Light Adaptation ◽  
Spectral Characteristics ◽  
Reversal Potential ◽  
Input Resistance ◽  
Postsynaptic Membrane ◽  
Resting Potential ◽  
Omega 3 ◽  
Visual Interneuron

1. We studied graded synaptic transmission in the fly photoreceptor-interneuron synapse by using intracellular in situ recordings from pre- and postsynaptic cells. 2. A large presynaptic hyperpolarization after light adaptation, caused by the activation of the electrogenic Na+/K+ pump, drastically reduced the conspicuous postsynaptic dark noise. At the same time, the postsynaptic neurons depolarized, with an increase of input resistance of 5-10 M omega. 3. The spectral characteristics of the postsynaptic membrane noise in dark and during noise reduction, together with the other results, suggested that the transmitter release decreased dramatically approximately 12 mV below the resting potential of the presynaptic photoreceptors. 4. During the postsynaptic noise reduction, the saturated and subsaturated first-order visual interneuron responses were increased up to 9 mV with a time constant of recovery of approximately 10 s. This increase was shown to be caused by the negative shift of the reversal potential of the transmitter-gated (mainly Cl-) conductance, caused apparently by the reduced transmitter input. 5. The results strongly suggest that the photoreceptor transmitter release in fly is tonic, even in dark, and further support the modulation of the synaptic voltage transfer by postsynaptic Cl- extrusion.

scholarly journals Botulinum Neurotoxin a Blocks Synaptic Vesicle Exocytosis but Not Endocytosis at the Nerve Terminal

1999 ◽  
Vol 147 (6) ◽  
pp. 1249-1260 ◽  
Author(s):  
Elaine A. Neale ◽  
Linda M. Bowers ◽  
Min Jia ◽  
Karen E. Bateman ◽  
Lura C. Williamson
Keyword(s):  
Synaptic Vesicle ◽  
Nerve Terminal ◽  
Neurotransmitter Release ◽  
Synaptic Vesicles ◽  
Botulinum Neurotoxin ◽  
Vesicle Membrane ◽  
Botulinum Neurotoxin A ◽  
Synaptic Vesicle Exocytosis ◽  
Vesicle Exocytosis ◽  
Botulinum Neurotoxins

The supply of synaptic vesicles in the nerve terminal is maintained by a temporally linked balance of exo- and endocytosis. Tetanus and botulinum neurotoxins block neurotransmitter release by the enzymatic cleavage of proteins identified as critical for synaptic vesicle exocytosis. We show here that botulinum neurotoxin A is unique in that the toxin-induced block in exocytosis does not arrest vesicle membrane endocytosis. In the murine spinal cord, cell cultures exposed to botulinum neurotoxin A, neither K+-evoked neurotransmitter release nor synaptic currents can be detected, twice the ordinary number of synaptic vesicles are docked at the synaptic active zone, and its protein substrate is cleaved, which is similar to observations with tetanus and other botulinal neurotoxins. In marked contrast, K+ depolarization, in the presence of Ca2+, triggers the endocytosis of the vesicle membrane in botulinum neurotoxin A–blocked cultures as evidenced by FM1-43 staining of synaptic terminals and uptake of HRP into synaptic vesicles. These experiments are the first demonstration that botulinum neurotoxin A uncouples vesicle exo- from endocytosis, and provide evidence that Ca2+ is required for synaptic vesicle membrane retrieval.

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