Cooperative Glutamatergic and Cholinergic Mechanisms Generate Short-Term Modifications of Synaptic Effectiveness in Prepositus Hypoglossi Neurons

To maintain horizontal eye position on a visual target after a saccade, extraocular motoneurons need a persistent (tonic) neural activity, called "eye-position signal," generated by prepositus hypoglossi (PH) neurons. We have shown previously in vitro and in vivo that this neural activity depends, among others mechanisms, on the interplay of glutamatergic transmission and cholinergic synaptically triggered depolarization. Here, we used rat sagittal brainstem slices, including PH nucleus and paramedian pontine reticular formation (PPRF). We made intracellular recordings of PH neurons and studied their synaptic activation from PPRF neurons. Train stimulation of the PPRF area evoked a cholinergic-sustained depolarization of PH neurons that outlasted the stimulus. EPSPs evoked in PH neurons by single pulses applied to the PPRF presented a short-term potentiation (STP) after train stimulation. APV (an NMDA-receptor blocker) or chelerythrine (a protein kinase-C inhibitor) had no effect on the sustained depolarization, but they did block the evoked STP, whereas pirenzepine (an M1 muscarinic antagonist) blocked both the sustained depolarization and the STP of PH neurons. Thus, electrical stimulation of the PPRF area activates both glutamatergic and cholinergic axons terminating in the PH nucleus, the latter producing a sustained depolarization probably involved in the genesis of the persistent neural activity required for eye fixation. M1-receptor activation seems to evoke a STP of PH neurons via NMDA receptors. Such STP could be needed for the stabilization of the neural network involved in the generation of position signals necessary for eye fixation after a saccade

A Cholinergic Synaptically Triggered Event Participates in the Generation of Persistent Activity Necessary for Eye Fixation

An exciting topic regarding integrative properties of the nervous system is how transient motor commands or brief sensory stimuli are able to evoke persistent neuronal changes, mainly as a sustained, tonic action potential firing. A persisting firing seems to be necessary for postural maintenance after a previous movement. We have studied in vitro and in vivo the generation of the persistent neuronal activity responsible for eye fixation after spontaneous eye movements. Rat sagittal brainstem slices were used for the intracellular recording of prepositus hypoglossi (PH) neurons and their synaptic activation from nearby paramedian pontine reticular formation (PPRF) neurons. Single electrical pulses applied to the PPRF showed a monosynaptic glutamatergic projection on PH neurons, acting on AMPA-kainate receptors. Train stimulation of the PPRF area evoked a sustained depolarization of PH neurons exceeding (by hundreds of milliseconds) stimulus duration. Both duration and amplitude of this sustained depolarization were linearly related to train frequency. The train-evoked sustained depolarization was the result of interaction between glutamatergic excitatory burst neurons and cholinergic mesopontine reticular fibers projecting onto PH neurons, because it was prevented by slice superfusion with cholinergic antagonists and mimicked by cholinergic agonists. As expected, microinjections of cholinergic antagonists in the PH nucleus of alert behaving cats evoked a gaze-holding deficit consisting of a re-centering drift of the eye after each saccade. These findings suggest that a slow, cholinergic, synaptically triggered event participates in the generation of persistent activity characteristic of PH neurons carrying eye position signals

Neurons of the Dentate Molecular Layer in the Rabbit Hippocampus

The molecular layer of the dentate gyrus appears as the main entrance gate for information into the hippocampus, i.e., where the perforant path axons from the entorhinal cortex synapse onto the spines and dendrites of granule cells. A few dispersed neuronal somata appear intermingled in between and probably control the flow of information in this area. In rabbits, the number of neurons in the molecular layer increases in the first week of postnatal life and then stabilizes to appear permanent and heterogeneous over the individuals’ life span, including old animals. By means of Golgi impregnations, NADPH histochemistry, immunocytochemical stainings and intracellular labelings (lucifer yellow and biocytin injections), eight neuronal morphological types have been detected in the molecular layer of developing adult and old rabbits. Six of them appear as interneurons displaying smooth dendrites and GABA immunoreactivity: those here called as globoid, vertical, small horizontal, large horizontal, inverted pyramidal and polymorphic. Additionally there are two GABA negative types: the sarmentous and ectopic granular neurons. The distribution of the somata and dendritic trees of these neurons shows preferences for a definite sublayer of the molecular layer: small horizontal, sarmentous and inverted pyramidal neurons are preferably found in the outer third of the molecular layer; vertical, globoid and polymorph neurons locate the intermediate third, while large horizontal and ectopic granular neurons occupy the inner third or the juxtagranular molecular layer. Our results reveal substantial differences in the morphology and electrophysiological behaviour between each neuronal archetype in the dentate molecular layer, allowing us to propose a new classification for this neural population

Role Of Neuropeptides In Migraine: Where Do They Stand In The Latest Expert Recommendations In Migraine Treatment?

Many factors have been implicated in the pathogenesis of migraine headache, including activation of the trigeminovascular system, dysfunction of: cerebral blood vessels, circulating vasoactive substances, mitochondrial energy metabolism, brain oxygenation and metabolism, platelet disorder, alterations in serotonin levels, low levels of brain tissue magnesium, altered transport of ions across the cell membrane, and inheritance and dysfunction of the brainstem periaqueductal gray matter. The headache phase of migraine is associated with cerebral vasodilation and inflammation, presumably mediated by the release of vasoactive substances and neuropeptides including CGRP (calcitonin gene-related peptide). Increased serum CCRP levels have been detected during migraine and cluster headache. One strategy to treat migraine is to inhibit the release of neuropeptides or to block their receptors. This article briefly reviews some experimental and clinical investigations focused on neuropeptide involvement in migraine. © 2007 Wiley-Liss, Inc

3-hydroxi-anthranilic acid is early expressed in stroke

<p>Using an immunohistochemical technique, we have studied the distribution of 3-OH-anthranilic acid (3-HAA) in the rat brain. Our study was carried out in control animals and in rats in which a stroke model (single transient middle cerebral artery occlusion) was performed. A monoclonal antibody directed against 3-HAA was also developed. 3-HAA was exclusively observed in the infarcted regions (ipsilateral striatum/cerebral cortex), 2, 5 and 21 days after the induction of stroke. In control rats and in the contralateral side of the stroke animals, no immunoreactivity for 3-HAA was visualized. Under pathological conditions (from early phases of stroke), we reported for the first time the presence of 3-HAA in the mammalian brain. By double immunohistochemistry, the coexistence of 3-HAA and GFAP was observed in astrocytes. The distribution of 3-HAA matched perfectly with the infarcted regions. Our findings suggest that, in stroke, 3-HAA could be involved in the tissue damage observed in the infarcted regions, since it is well known that 3-HAA exerts cytotoxic effects.</p

NO-tryptophan: a new small molecule located in the rat brain

<p>A highly specific monoclonal antibody directed against nitric oxide-tryptophan (NO-W) with good affinity (10^-9M) and specificity was developed. In the rat brain, using an indirect immunoperoxidase technique, cell bodies containing NO-W were exclusively found in the intermediate and dorsal parts of the lateral septal nucleus. No immunoreactive fibres were found in the rat brain. This work reports the first visualization and the morphological characteristics of cell bodies containing NO-W in the mammalian brain. The restricted distribution of NO-W in the rat brain suggests that this molecule could be involved in specific physiological mechanisms. </p

Muscarinic activation of a non-selective cationic conductance in pyramidal neurons in rat basolateral amygdala

In the present study, a cationic membrane conductance activated by the acetylcholine agonist carbachol was characterized in vitro in neurons of the basolateral amygdala. Extracellular perfusion of the K+ channel blockers Ba2+ and Cs+ or loading of cells with cesium acetate did not affect the carbachol-induced depolarization. Similarly, superfusion with low-Ca2+ solution plus Ba2+ and intracellular EGTA did not affect the carbachol-induced depolarization, suggesting a Ca2+-independent mechanism. On the other hand, the carbachol-induced depolarization was highly sensitive to changes in extracellular K+ or Na+. When the K+ concentration in the perfusion medium was increased from 4.7 to 10 mM, the response to carbachol increased in amplitude. In contrast, lowering the extracellular Na+ concentration from 143.2 to 29 mM abolished the response in a reversible manner. Results of coapplication of carbachol and atropine, pirenzepine or gallamine indicate that the carbachol-induced depolarization was mediated by muscarinic cholinergic receptors, but not the muscarinic receptor subtypes M1, M2 or M4, specifically. These data indicate that, in addition to the previously described reduction of a time- and voltage-independent K+ current (IKleak), a voltage- and time-dependent K+ current (IM), a slow Ca2+-activated K+ current (sIahp) and the activation of a hyperpolarization-activated inward rectifier K+ current (IQ), carbachol activated a Ca2+-independent non-selective cationic conductance that was highly sensitive to extracellular K+ and Na+ concentrations

Muscarinic activation of a non-selective cationic conductance in pyramidal neurons in rat basolateral amygdala

In the present study, a cationic membrane conductance activated by the acetylcholine agonist carbachol was characterized in vitro in neurons of the basolateral amygdala. Extracellular perfusion of the K+ channel blockers Ba2+ and Cs+ or loading of cells with cesium acetate did not affect the carbachol-induced depolarization. Similarly, superfusion with low-Ca2+ solution plus Ba2+ and intracellular EGTA did not affect the carbachol-induced depolarization, suggesting a Ca2+-independent mechanism. On the other hand, the carbachol-induced depolarization was highly sensitive to changes in extracellular K+ or Na+. When the K+ concentration in the perfusion medium was increased from 4.7 to 10 mM, the response to carbachol increased in amplitude. In contrast, lowering the extracellular Na+ concentration from 143.2 to 29 mM abolished the response in a reversible manner. Results of coapplication of carbachol and atropine, pirenzepine or gallamine indicate that the carbachol-induced depolarization was mediated by muscarinic cholinergic receptors, but not the muscarinic receptor subtypes M1, M2 or M4, specifically. These data indicate that, in addition to the previously described reduction of a time- and voltage-independent K+ current (IKleak), a voltage- and time-dependent K+ current (IM), a slow Ca2+-activated K+ current (sIahp) and the activation of a hyperpolarization-activated inward rectifier K+ current (IQ), carbachol activated a Ca2+-independent non-selective cationic conductance that was highly sensitive to extracellular K+ and Na+ concentrations

Muscarinic agonist carbachol depresses excitatory synaptic transmission in the rat basolateral amygdala in vitro.

Intracellular recordings in slice preparations of the basolateral amygdala were used to test which excitatory amino acid receptors mediate the excitatory postsynaptic potentials due to stimulation of the external capsule. These recordings were also used to examine the action of muscarinic agonists on the evoked excitatory potentials. Intracellular recordings from amygdaloid pyramidal neurons revealed that carbachol (2-20 microM) suppressed, in a dose-dependent manner, excitatory postsynaptic responses evoked by stimulation of the external capsule (EC). This effect was blocked by atropine. The estimated effective concentration to produce half-maximal response (EC(50)) was 6.2 microM. Synaptic suppression was observed with no changes in the input resistance of the recorded cells, suggesting a presynaptic mechanism. In addition, the results obtained using the paired-pulse protocol provided additional support for a presynaptic action of carbachol. To identify which subtype of cholinergic receptors were involved in the suppression of the EPSP, four partially selective muscarinic receptor antagonists were used at different concentrations: pirenzepine, a compound with a similar high affinity for muscarinic M1 and M4 receptors; gallamine, a noncompetitive antagonist for M2; methoctramine, an antagonist for M2 and M4; and 4-diphenylacetoxy-N-methylpiperidine, a compound with similar high affinity for muscarinic receptors M1 and M3. None of them independently antagonized the suppressive effect of carbachol on the evoked EPSP completely, suggesting that more than one muscarinic receptor subtype is involved in the effect. These experiments provide evidence that in the amygdala muscarinic agonists block the excitatory synaptic response, mediated by glutamic acid, by acting on several types of presynaptic receptors

Randomized controlled trial comparing the effectiveness of the ultrasound-guided galvanic electrolysis technique (USGET) versus conventional electro-physiotherapeutic treatment on patellar tendinopathy

Patellar tendinopathy has a high prevalence rate among athletes. Different therapeutic options can be found in the current literature, but none of them has been clearly established as the gold standard. The purpose of this study is to compare, in a randomized controlled trial, the clinical efficacy of eccentric exercise combined with either an ultrasound-guided galvanic electrolysis technique (USGET) or conventional electrophysiotherapy to treat patellar tendinopathy. Sixty patients diagnosed with patellar tendinopathy were randomized into two groups. Group 1 (n = 30) received electrophysiotherapy treatment consisting of ultrasound, laser and interferential current techniques. Group 2 (n = 30) received USGET. Both groups did the same standardized eccentric exercise program. Periodic assessments of the subjects were carried out with the Victorian Institute of Sport Assessment-Patella (VISA-P) score. An analysis of means and a survival study were performed. There were statistically significant differences in the VISA-P between the baseline and final follow-up in each treatment group. Group 1 (conventional electrophysiotherapy) went from 52.5 ± 18.8 to 61.9 ± 13.7 (in VISA-P 90 subgroup). Group 2 (USGET) went from 51.4 ± 17.9 to 63.3 ± 14.3 (in VISA-P 90 subgroup). There were statistically significant correlations between the baseline and final score in the VISA-P > 90 subjects upon completing the study but no statistically significant correlations between subjects with VISA-P < 90. The mean number of sessions applied was 22.6 ± 2.5 in Group 1 and 3.2 ± 0.9 in Group 2. The success probability in Group 1 was 36.1% versus 72.4% in Group 2. The difference was statistically significant. The results obtained with the combination of USGET and eccentric exercise reported better outcomes than with the conventional electrophysiotherapy techniques in the treatment of patellar tendinopathy