The analysis of quantal characteristics of spontaneous evoked excitatory post synaptic potential sin long-term depression of glutamatergic neurotransmission between cultured hippocampal neurons

The results of electrophysiological studies of the quantal release features of glutamate in hippocampal neurons' synapses in long-term depression of synaptic transmission are presented. To research the topic, weused the primary culture of the rat hippocampus. Using the patch-clamp technique, the frequencies and amplitudes of excitatory spontaneous postsynaptic currents (EPSCs) were determined, theirdistributions were analyzed and the basicquantal parameters were calculated. Long-term depression of synaptic transmission was achieved by tetanic stimulation of the axon of the presynaptic cell for 5 minutes with a stimulation frequency of 5 Hz. Spontaneous current in depression was recorded 20-30 min after tetanic stimulation. It has been shown that the amplitude of the EPSCs registered in hippocampal neurons decreased in the long-term depression of synaptic transmission. It has been found that two neurotransmitter vesicles are usually released in the control at the same time and one during the long-term depression of synaptic transmission.It means that the probability of release decreased. In addition, the average quantalamplitude and quantum content decreased. It indicates the effect of presynaptic mechanisms in the expression of long-term depression of synaptic neurotransmission between hippocampal neurons in culture.In the conditions of long-term depression, no significant changes were found in the frequency of EPSCs. The analysis of quantum parameters in long-term depression of glutamatergic synaptic transmission between neurons is important for the formation of more complete ideas about the mechanisms that play a fundamental role in the normal functioning of the CNS and the development of neural networks.

Developmental synaptic changes at the transient olivocochlear-inner hair cell synapse

In the mature mammalian cochlea, inner hair cells (IHCs) are mainly innervated by afferent fibers that convey sound information to the central nervous system. During postnatal development, however, medial olivocochlear (MOC) efferent fibers transiently innervate the IHCs. The MOC-IHC synapse, functional from postnatal day (P)0 to hearing onset (P12), undergoes dramatic changes in the sensitivity to acetylcholine (ACh) and in the expression of key postsynaptic proteins. To evaluate whether there are associated changes in the properties of ACh release during this period, we used a cochlear preparation from mice at P4, P6-7 and P9-11 and monitored transmitter release from MOC terminals in voltage-clamped IHCs in the whole-cell configuration. The quantum content increased 5.6x from P4 to P9-11 due to increases in the size and replenishment rate of the readily releasable pool (RRP) of synaptic vesicles, without changes in their probability of release (Pvesicle) or quantum size. This strengthening in transmission was accompanied by changes in the short-term plasticity (STP) properties, which switched from facilitation at P4 to depression at P9-11. We have previously shown that at P9-11, ACh release is supported by P/Q and N-type voltage-gated calcium channels (VGCCs) and negatively regulated by BK potassium channels activated by Ca2+ influx through L-type VGCCs. We now show that at P4 and P6-7, release is mediated by P/Q-, R- and L-type VGCCs. Interestingly, L-type VGCCs have a dual role: they both support release and fuel BK channels, suggesting that at immature stages the presynaptic proteins involved in release are less compartmentalized.Significance statementDuring postnatal development prior to the onset of hearing, cochlear IHCs present spontaneous Ca2+ action potentials which release glutamate at the first auditory synapse in the absence of sound stimulation. The IHC Ca2+ action potential frequency pattern, which is crucial for the correct establishment and function of the auditory system, is regulated by the efferent MOC system that transiently innervates IHCs during this period. We show short-term synaptic plasticity properties of the MOC-IHC synapse that tightly shape this critical developmental period.

THE QUANTUM CONTENT OF THE NORMAL SURFACES IN A THREE-MANIFOLD

The formula for the Turaev–Viro invariant of a three-manifold depends on a complex parameter t. When t is not a root of unity, the formula becomes an infinite sum. This paper analyzes convergence of this sum when t does not lie on the unit circle, in the presence of an efficient triangulation of the three-manifold. The terms of the sum can be indexed by surfaces lying in the three-manifold. The contribution of a surface is largest when the surface is normal and when its genus is the lowest.

Neuregulin Effect on Quantal Content Dissociated From Effect on Miniature Endplate Potential Amplitude

Members of the neuregulin family of signaling proteins increase transcription of acetylcholine receptor (AChR) subunit genes in muscle fibers and the number of AChRs in the muscle membrane. In adult mice heterozygous for targeted deletion of type I neuregulins (Ig-NRG+/−), postsynaptic AChR density was decreased and transmitter release was increased. We examined the relationship between functional AChR density and ACh release in postnatal day 7 (P7), P14, and adult NRG-deficient mice. Here we report that changes in postsynaptic sensitivity and transmitter release are not temporally coupled during postnatal development in Ig-NRG–deficient mice. Although miniature endplate potential (MEPP) amplitude was decreased compared with control in P7 Ig-NRG+/− mice, quantum content was not increased. Quantum content was increased in adult heterozygotes despite normal MEPP amplitudes. Thus, during postnatal maturation, both quantal size and quantum content were influenced by decreased Ig-NRG expression, although the effects were dissociated in time.

Histamine Compartments of the Drosophila Brain With an Estimate of the Quantum Content at the Photoreceptor Synapse

Reliable estimates of the quantum size in histaminergic neurons are not available. We have exploited two unusual opportunities in the fly's ( Drosophila melanogaster) visual system to make such determinations for histaminergic photoreceptor synapses: 1) the possibility to microdissect successively from whole fly heads freeze-dried in acetone: the compound eyes; the first optic neuropils, or lamina; and the rest of the brain; and 2) the uniform sheaves of lamina synaptic terminals of photoreceptors R1–R6. We used this organization to count scrupulously the numbers of 30-nm synaptic vesicles from electron micrographs of R1–R6 profiles, and from microdissections we determined the regional contents of histamine in the compound eye, lamina, and central brain. Total head histamine averages 1.98 ng of which 9% was lost after freeze-drying in acetone and a further 28% after the brain was microdissected. Of the remainder, 71% was in the eye and lamina. Assuming that histamine loss from the tissue occurred mostly by diffusion evenly distributed among all regions, the overall lamina content of the head would be 0.1935 ng before dissection. From published values for the volumes of the brain's compartments, the computed regional concentrations of histamine are highest in the lamina (4.35 mM) because of the terminals of R1–R6. The concentration in the retina is ∼13% that in the lamina, suggesting that most histamine is vesicular. There are ∼43,500 ± 7,400 (SD) synaptic vesicles per terminal and, if all histamine is allocated equally and exclusively among these, the vesicle contents would be 858 ± 304 × 10−21 moles or ∼5,000 ± 1,800 (SD) molecules at an approximate concentration of 670 mM. These values are compared with the vesicle contents at synapses using acetylcholine and catecholamines.

Neuromuscular transmission in dystrophic mice

1. Neuromuscular transmission was studied in the extensor digitorum-longus muscle of dystrophic mice (strain 129/ReJ) by means of intracellular recording techniques. 2. In a large population of normal and dystrophic muscle fibers tested, the incidence of transmission failure was about 2% and showed no significant difference between the two groups. 3. Quantal size and quantum content of dystrophic junctions were found to be normal. This was true even of nerve terminal on apparently atrophied muscle fibers. 4. The facilitation ratio at dystrophic junctions was not significantly different from normal. 5. Dystrophic neuromuscular junctions exhibited an abnormality high frequency of giant spontaneous potentials. Application of tetrodotoxin (10(-6) M) and curare (10(-6) M) indicated that these potentials were caused by impulse-independent release of acetylcholine. 6. Neuromuscular transmission in dystrophic mice was found functionally normal and unrelated to the degenerative state of the muscle.

Morphological correlates of synaptic transmission in lamprey spinal cord

The dye Procion brown was used to identify in the light and electron microscope, synaptic contacts made between monosynaptically coupled neurons in the lamprey spinal cord whose synaptic interaction had been recorded. Synaptic contacts were made on different dendrites of the postsynaptic cell at different distances from the soma. Some of the contacts were made on dentritic spines and some on the smooth shaft of the dentrites. Serial sections through synaptic contacts made on dendritic processess of the postsynaptic cells were used for three-dimensional reconstruction of the synapses using computer graphics techniques. The computer reconstructions and detailed examination of the serial EM micrographs revealed the large proliferation of membrane involved in making these en passant synapses as well as the morphological changes due to stimulation of the presynaptic axon. These changes include depletion of synaptic vesicles and formation of complex vesicles and synaptic cisternae. Besides chemical synaptic contacts, four electrotonic contacts were located, confirming the mixed electrochemical synaptic response recorded from the postsynaptic cell. The mean quantum content was estimated and compared with the estimate of the available transmitter pool, assuming the quantal release hypothesis applies at these synapses. The total transmitter pool was estimated by counting all synaptic vesicles in all synaptic contacts. It was estimated that about 6% of the total transmitter pool is available for release at these synapses. This compares with less than 1% at the neuromuscular junction and about 20% at sympathetic synapses. These results support the hypothesis that synaptic vesicles may be recycled as described by Heuser and Reese (22) at the neuromuscular junction. Ongoing studies are investigating the effect on a variety of synaptic junctions to stimulation for different periods of time of presynaptic axons. The methods described in this study can also be used to test the models of synaptic interaction on dendritic trees described by Rall (39) and Jack and Redman (24).

Quantal Mechanism of Transmitter Release during Progressive Depletion of the Presynaptic Stores at a Ganglionic Synapse

In the present experiments we interfered with the mechanism of acetylcholine (ACh) synthesis in the rat superior cervical ganglion by impairing the supply of either the choline group (hemicholinium no. 3 [HC-3]treatment) or the acetyl group (thiamine deprivation). Under both conditions stimulation causes in the ganglion a progressive decline in ACh output associated with a depletion of transmitter tissue content. ACh release from the terminals of a single preganglionic fiber was estimated from the quantum content value of the evoked excitatory postsynaptic potentials (EPSP's) recorded intracellularly in the ganglion neuron under test. The present observations indicate that Poisson statistics describe transmitter release at either low or high release levels. Furthermore, the progressive decline in the rate of ACh output occurring during repetitive stimulation is shown to correspond to a progressive decrease in the number of transmitter quanta released per impulse and not to any modification in the size of individual quanta. Some 8,000 transmitter quanta proved to represent the presynaptic transmitter store initially present in those terminals on a neuron that are activated by stimulation of a single preganglionic fiber. Speculations are considered about synaptic efficacy and nerve connections in rat autonomic ganglia. It is suggested that six preganglionic fibers represent the mean input to a ganglion neuron.