Prolonged High Frequency Stimulation Research Articles

CB1 cannabinoid receptor-mediated plasticity of GABAergic synapses in the mouse insular cortex

The insular cortex plays pivotal roles in taste learning. As cellular mechanisms of taste learning, long-term potentiation (LTP) at glutamatergic synapses is well studied. However, little is known about long-term changes of synaptic efficacy at GABAergic synapses in the insular cortex. Here, we examined the synaptic mechanisms of long-term plasticity at GABAergic synapses in layer V pyramidal neurons of the mouse insular cortex. In response to a prolonged high-frequency stimulation (HFS), GABAergic synapses displayed endocannabinod (eCB)-mediated long-term depression (LTDGABA). When cannabinoid 1 receptors (CB1Rs) were blocked by a CB1R antagonist, the same stimuli caused LTP at GABAergic synapses (LTPGABA) which was mediated by production of nitric oxide (NO) via activation of NMDA receptors. Intriguingly, NO signaling was necessary for the induction of LTDGABA. In the presence of leptin which blocks CB1 signaling, the prolonged HFS caused LTPGABA which was mediated by NO signaling. These results indicate that long-term plasticity at GABAergic synapses in the insular cortex can be modulated by combined effects of eCB and NO signaling. These forms of GABAergic synaptic plasticity in the insular cortex may be crucial synaptic mechanisms in taste learning.

High frequency stimulation of afferent fibers generates asynchronous firing in the downstream neurons in hippocampus through partial block of axonal conduction

Deep brain stimulation (DBS) is effective for treating neurological disorders in clinic. However, the therapeutic mechanisms of high-frequency stimulation (HFS) of DBS have not yet been elucidated. Previous studies have suggested that HFS-induced changes in axon conduction could have important contributions to the DBS effects and desiderate further studies. To investigate the effects of prolonged HFS of afferent axons on the firing of downstream neurons, HFS trains of 100 and 200Hz were applied on the Schaffer collaterals of the hippocampal CA1 region in anaesthetized rats. Single unit activity of putative pyramidal cells and interneurons in the downstream region was analyzed during the late periods of prolonged HFS when the axonal conduction was blocked. The results show that the firing rates of both pyramidal cells and interneurons increased rather than decreased during the period of axon block. However, the firing rates were far smaller than the stimulation frequency of HFS. In addition, the firing pattern of pyramidal cells changed from typical bursts during baseline recordings into regular single spikes during HFS periods. Furthermore, the HFS produced asynchronous firing in the downstream neurons in contrast to the synchronous firing induced by single pulses. Presumably, the HFS-induced block of axonal conduction was not complete. During the period of partial block, individual axons could recover intermittently and independently, and drive the downstream neurons to fire in an asynchronous pattern. This axonal mechanism of HFS provides a novel explanation for how DBS could replace an original pattern of neuronal activity by a HFS-modulated asynchronous firing in the target region thereby generating the therapeutic effects of DBS.

Tumor necrosis factor-α potentiates long-term potentiation in the rat dentate gyrus after acute hypoxia.

An inadequate supply of oxygen in the brain may lead to an inflammatory response through neuronal and glial cells that can result in neuronal damage. Tumor necrosis factor-α (TNF-α) is a proinflammatory cytokine that is released during acute hypoxia and can have neurotoxic or neuroprotective effects in the brain. Both TNF-α and interleukin-1β (IL-1β) have been shown by a number of research groups to alter synaptic scaling and also to inhibit long-term potentiation (LTP) in the hippocampus when induced by specific high-frequency stimulation (HFS) protocols. This study examines the effects of TNF-α on synaptic transmission and plasticity in hippocampal slices after acute hypoxia using two HFS protocols. Field excitatory postsynaptic potentials were elicited in the medial perforant pathway of the dentate gyrus. Exogenous TNF-α (5 ng/ml) attenuated LTP induced by theta burst stimulation but had no effect on LTP induced by a more prolonged HFS. Pretreatment with lipopolysaccharide (100 ng/ml) or TNF-α but not IL-1β (4 ng/ml) prior to a 30-min hypoxic insult resulted in a significant enhancement of LTP post hypoxia when induced by the HFS. Anti-TNF, 3,6'-dithiothalidomide (a TNF-α synthesis inhibitor), and SB203580 (a p38 MAPK inhibitor) significantly reduced this effect. These results indicate an important modulatory role for elevated TNF-α levels on LTP in the hippocampus after an acute hypoxic event.

Temperature dependence of vesicular dynamics at excitatory synapses of rat hippocampus.

How vesicular dynamics parameters depend on temperature and how temperature affects the parameter change during prolonged high frequency stimulation was determined by fitting a model of vesicular storage and release to the amplitudes of the excitatory post-synaptic currents (EPSC) recorded from CA1 neurons in rat hippocampal slices. The temperature ranged from low (13°C) to higher and more physiological temperature (34°C). Fitting the model of vesicular storage and release to the EPSC amplitudes during a single pair of brief high-low frequency stimulation trains yields the estimates of all parameters of the vesicular dynamics, and with good precision. Both fractional release and replenishment rate decrease as the temperature rises. Change of the underlying 'basic' parameters (release coupling, replenishment coupling and readily releasable pool size), which the model-fitting also yields is complex. The replenishment coupling between the readily releasable pool (RRP) and resting pool increases with temperature (which renders the replenishment rate higher), but this is more than counterbalanced by greater RRP size (which renders the replenishment rate lower). Finally, during long, high frequency patterned stimulation that leads to significant synaptic depression, the replenishment rate decreases markedly and rapidly at low temperatures (<22°C), but at high temperatures (>28°C) the replenishment rate rises with stimulation, making synapses better able to maintain synaptic efficacy.

Changes in Kinetics of Calcium Signals in Response to Prolonged High Frequency Stimulation of Cultured Hippocampal Neurons

Changes in Kinetics of Calcium Signals in Response to Prolonged High Frequency Stimulation of Cultured Hippocampal Neurons

Mutagenesis Studies Investigating Slow Inactivation of Sodium Channel Nav1.6

Slow inactivation (SI) determines the availability of voltage-gated sodium channels and effects action potential firing patterns. Neuronally-expressed Nav1.6 was suggested to play an important role in maintaining high firing frequencies, possibly due to a reduced SI compared to other sodium channel subtypes. With this study we characterized SI of WT Nav1.6R and introduced mutations that severely altered the channel's propensity to slow inactivate. Conserved residues in the S6 segment of DII and DIII were identified as SI-sensitive hot spots: V966C in DII and N1455A in DIII. WT and mutant channels were transiently expressed in ND7/23 cells and patch-clamped. The N1455A mutation greatly enhanced SI whereas the V966C mutation shifted its voltage-dependence in the depolarizing direction by ∼25 mV. Fast inactivation was shifted slightly to more hyperpolarized potentials (∼3 mV for V966C and ∼8.5 mV for N1455A). We applied trains of depolarizing stimuli to imitate high-frequency firing. At a stimulation frequency of 50 and 100 Hz, only the N1455A mutation displayed enhanced run down, whereas V966C behaved similar to WT, probably due to the slight enhancement of fast inactivation of V966C. Our findings show that Nav1.6 SI can be severely altered by mutations in DII and DIII S6 segments, thereby confirming the importance of these regions to SI. N1455A induces loss of Nav1.6 during prolonged high frequency stimulation, which will allow us to test for the specific role of Nav1.6 vs. Nav1.7 in action potential (AP) generation: when transfected into DRGs, N1455A may attenuate prolonged high-frequency firing, whereas the threshold (thought to be set by Nav1.7) may remain unaffected. With this mutations we produced a useful molecular tool and approach for elucidating the role and influence of Nav1.6 SI on firing patterns in sensory neurons.

Metabotropic glutamate receptor-mediated use-dependent down-regulation of synaptic excitability involves the fragile X mental retardation protein.

Loss of the mRNA-binding protein FMRP results in the most common inherited form of both mental retardation and autism spectrum disorders: fragile X syndrome (FXS). The leading FXS hypothesis proposes that metabotropic glutamate receptor (mGluR) signaling at the synapse controls FMRP function in the regulation of local protein translation to modulate synaptic transmission strength. In this study, we use the Drosophila FXS disease model to test the relationship between Drosophila FMRP (dFMRP) and the sole Drosophila mGluR (dmGluRA) in regulation of synaptic function, using two-electrode voltage-clamp recording at the glutamatergic neuromuscular junction (NMJ). Null dmGluRA mutants show minimal changes in basal synapse properties but pronounced defects during sustained high-frequency stimulation (HFS). The double null dfmr1;dmGluRA mutant shows repression of enhanced augmentation and delayed onset of premature long-term facilitation (LTF) and strongly reduces grossly elevated post-tetanic potentiation (PTP) phenotypes present in dmGluRA-null animals. Null dfmr1 mutants show features of synaptic hyperexcitability, including multiple transmission events in response to a single stimulus and cyclic modulation of transmission amplitude during prolonged HFS. The double null dfmr1;dmGluRA mutant shows amelioration of these defects but does not fully restore wildtype properties in dfmr1-null animals. These data suggest that dmGluRA functions in a negative feedback loop in which excess glutamate released during high-frequency transmission binds the glutamate receptor to dampen synaptic excitability, and dFMRP functions to suppress the translation of proteins regulating this synaptic excitability. Removal of the translational regulator partially compensates for loss of the receptor and, similarly, loss of the receptor weakly compensates for loss of the translational regulator.

Differential effects of prolonged high frequency stimulation and of excitotoxic lesion of the subthalamic nucleus on dopamine denervation‐induced cellular defects in the rat striatum and globus pallidus

This study examined the effects of prolonged (4 days) high frequency stimulation (HFS) of the subthalamic nucleus (STN), in comparison with those of STN lesion, on the dopamine denervation-mediated cellular changes in the basal ganglia in a Wistar rat model of Parkinson's disease. STN HFS counteracted the dopamine lesion-induced increase in GAD67 mRNA expression in the output structures of the basal ganglia, as shown previously after STN lesion, providing cellular support for the similar antiparkinsonian benefits produced by the two surgical procedures. The dopamine denervation-induced increase in GAD67 mRNA levels in the globus pallidus was partially antagonized after HFS and totally reversed after ibotenate-induced STN lesion. The overexpression of striatal enkephalin mRNA tended to be further increased by HFS but was antagonized by STN lesion. The decrease in striatal substance P mRNA levels was affected neither by STN HFS nor lesion. As STN HFS for two hours was previously found not to interfere with the effects of dopamine lesion in the globus pallidus and striatum, the present data provide strong evidence that the effects of STN surgery in these structures involve long-term adaptive processes and that the rearrangements mediated by HFS and lesion are, at least in part, different.

Changes in the Asynchronicity of Transmitter Release at the Neuromuscular Junction during Prolonged High-Frequency Stimulation

In experiments on neuromuscular junctions in the frog m. cutaneous-pectoris, changes in the intensity and asynchronicity of transmitter release during high-frequency (10 and 50 sec-1) rhythmic stimulation of the motor nerve were investigated using extracellular recording. At low extracellular Ca2+ concentrations, rhythmic stimulation resulted in a gradual enlargement of the quantum content of end-plate currents (EPC), the so-called facilitation. The latter phenomenon was accompanied by an increase in the average value and variance of synaptic delays of single-quantum EPC, a shift of the main mode of their distribution towards greater values, and an increase in the latency of the nerve ending responses. The above-described changes reduce the magnitude of facilitation in the neuromuscular synapse.

Phosphorylation and Local Presynaptic Protein Synthesis in Calcium- and Calcineurin-Dependent Induction of Crayfish Long-Term Facilitation

Long-term facilitation at the crayfish opener muscle is elicited by prolonged high frequency stimulation, and arises from an increase in functional active zones, resulting in increased transmitter release. LTF induction depends critically upon presynaptic calcium accumulation and calcineurin (PP2B) activity. The protein synthesis dependence of this synaptic strengthening was investigated. LTF occurred without transcription, but the translation inhibitors cycloheximide and anisomycin, or local presynaptic injection of mRNA cap analog m 7GpppG, impaired LTF expression. Both MAP kinase and phosphatidylinositol 3-OH kinase (PI3K) activation are implicated in this rapamycin-sensitive synaptic potentiation. This study defines an important role for protein synthesis in the expression of activity-dependent plasticity, and provides mechanistic insight for the induction of this process at presynaptic sites.

The innervation of salivary glands as revealed by morphological methods.

Salivary secretion is nerve mediated. The salivary glands are supplied by parasympathetic and sympathetic efferent nerves which travel to the glands by separate routes. Once in the glands the axons from each type of nerve intermingle and travel together in association with Schwann cells, forming Schwann-axon bundles. Two types of neuro-effector relationships exist with salivary parenchymal and myoepithelial cells: epilemmal (outside the parenchymal basement membrane) and hypolemmal (within the parenchymal basement membrane). Their relative frequencies with either type of nerve differ greatly between glands and species. Salivary blood vessels receive epilemmal innervations by both sympathetic and parasympathetic axons. The classical transmitters--acetylcholine in parasympathetic and noradrenaline in sympathetic axons--are stored in small vesicles. A variety of non-conventional neuropeptide transmitters have also been found in salivary nerves by immunohistochemistry, and they occur in large dense-cored vesicles. Prolonged high frequency stimulation has been found to cause depletion of large dense-cored vesicles from glandular nerves. In recent years afferent nerves have started to be identified and are found in greatest numbers around the main salivary ducts, where they may form a hypolemmal association with the epithelial cells. Functional studies demonstrate complex interactions between parasympathetic and sympathetic nerves. Morphological assessments of changes in the parenchymal cells after nerve stimulations or denervations add greatly to our understanding of the nerve functions. At least four types of influence can be exerted on salivary parenchymal cells by the nerves: hydrokinetic (water mobilizing), proteokinetic (protein secreting), synthetic (inducing synthesis), and trophic (maintaining normal functional size and state). In respect to each role, wide glandular and species differences exist between the relative contributions made by each type of nerve.