Differential Contribution of Ca2+-Dependent Mechanisms to Hyperexcitability in Layer V Neurons of the Medial Entorhinal Cortex.

Temporal lobe epilepsy is characterized by recurrent seizures in one or both temporal lobes of the brain; some in vitro models show that epileptiform discharges initiate in entorhinal layer V neurons and then spread into other areas of the temporal lobe. We previously found that, in the presence of GABAA receptor antagonists, stimulation of afferent fibers, terminating both at proximal and distal dendritic locations, initiated hyperexcitable bursts in layer V medial entorhinal neurons. We investigated the differential contribution of Ca2+-dependent mechanisms to the plateaus underlying these bursts at proximal and distal synapses. We found that the NMDA glutamatergic antagonist D,L-2-amino-5-phosphonovaleric acid (APV; 50 µM) reduced both the area and duration of the bursts at both proximal and distal synapses by about half. The L-type Ca2+ channel blocker nimodipine (10 µM) and the R- and T-type Ca2+ channel blocker NiCl2 (200 µM) decreased the area of the bursts to a lesser extent; none of these effects appeared to be location-dependent. Remarkably, the perfusion of flufenamic acid (FFA; 100 µM), to block Ca2+-activated non-selective cation currents (ICAN) mediated by transient receptor potential (TRP) channels, had a location-dependent effect, by abolishing burst firing and switching the suprathreshold response to a single action potential (AP) for proximal stimulation, but only minimally affecting the bursts evoked by distal stimulation. A similar outcome was found when FFA was pressure-applied locally around the proximal dendrite of the recorded neurons and in the presence of a selective blocker of melastatin TRP (TRPM) channels, 9-phenanthrol (100 µM), whereas a selective blocker of canonical TRP (TRPC) channels, SKF 96365, did not affect the bursts. These results indicate that different mechanisms might contribute to the initiation of hyperexcitability in layer V neurons at proximal and distal synapses and could shed light on the initiation of epileptiform activity in the entorhinal cortex.

Mouse ERG K(+) channel clones reveal differences in protein trafficking and function.

BACKGROUND: The mouse ether-a-go-go-related gene 1a (mERG1a, mKCNH2) encodes mERG K(+) channels in mouse cardiomyocytes. The mERG channels and their human analogue, hERG channels, conduct IKr. Mutations in hERG channels reduce IKr to cause congenital long-QT syndrome type 2, mostly by decreasing surface membrane expression of trafficking-deficient channels. Three cDNA sequences were originally reported for mERG channels that differ by 1 to 4 amino acid residues (mERG-London, mERG-Waterston, and mERG-Nie). We characterized these mERG channels to test the postulation that they would differ in their protein trafficking and biophysical function, based on previous findings in long-QT syndrome type 2. METHODS AND RESULTS: The 3 mERG and hERG channels were expressed in HEK293 cells and neonatal mouse cardiomyocytes and were studied using Western blot and whole-cell patch clamp. We then compared our findings with the recent sequencing results in the Welcome Trust Sanger Institute Mouse Genomes Project (WTSIMGP). CONCLUSIONS: First, the mERG-London channel with amino acid substitutions in regions of highly ordered structure is trafficking deficient and undergoes temperature-dependent and pharmacological correction of its trafficking deficiency. Second, the voltage dependence of channel gating would be different for the 3 mERG channels. Third, compared with the WTSIMGP data set, the mERG-Nie clone is likely to represent the wild-type mouse sequence and physiology. Fourth, the WTSIMGP analysis suggests that substrain-specific sequence differences in mERG are a common finding in mice. These findings with mERG channels support previous findings with hERG channel structure-function analyses in long-QT syndrome type 2, in which sequence changes in regions of highly ordered structure are likely to result in abnormal protein trafficking.

Effect of heterogeneity and noise on cross frequency phase-phase and phase-amplitude coupling.

Cross-frequency coupling is hypothesized to play a functional role in neural computation. We apply phase resetting theory to two types of cross-frequency coupling that can occur when a slower oscillator periodically forces one or more oscillators: phase-phase coupling, in which the two oscillations are phase-locked, and phase-amplitude coupling, in which the amplitude of the driven oscillation is modulated. Our first result is that the shape of the phase resetting curve predicts the tightness of locking to a pulsatile forcing periodic input at any ratio of forced to intrinsic period; the tightness of the locking decreases as the ratio increases. Theoretical expressions were obtained for the probability density of the phases for a population of heterogeneous oscillators or a noisy single oscillator. Results were confirmed using two types of simulated networks and experiments on hippocampal CA1 neurons. Theoretical expressions were also obtained and confirmed for the probability density of N spike times within a single cycle of low frequency forcing. The second result is a suggested mechanism for phase-amplitude coupling in which progressive desynchronization leads to decreasing amplitude during a low frequency forcing cycle. Network simulations confirmed the theoretical viability of this mechanism, and that it generalizes to more diffuse input.

Is commercially available point finder accurate and reliable in detecting active auricular acupuncture points?

OBJECTIVES: This study was done to determine the specificity and sensitivity of a commercial Pointer Plus (Point finder) in detecting a region of low skin resistance on the ear. DESIGN: This was a prospective blinded study. SETTING/LOCATION: The study was done at the Yale New Haven Hospital, New Haven, CT. SUBJECTS: The subjects were men and women who work at Yale New Haven Hospital. INTERVENTIONS: There were no interventions. OUTCOME MEASURES: Correlations were made between self-reported musculoskeletal pain and the detection of low skin resistance on the ear. RESULTS: The positive predictive value for Pointer Plus detecting low skin resistance correlating to the neck region of French auricular map is 0.76 (76%). The positive predictive value for Pointer Plus to detect low skin resistance area correlating to the low back region of French auricular map is 0.25. The positive predictive value for Pointer Plus in detecting any low in skin resistance on the external auricles in patients who complained of more than two musculoskeletal pains is 0.29. CONCLUSIONS: The specificity and sensitivity of a commercial Pointer Plus (point finder) in detecting a region of low skin resistance on the ear being unreliable, depending on the correlating area based on a published auricular map. Additional assessments are needed to support the clinical practice.

Properties of WT and mutant hERG K(+) channels expressed in neonatal mouse cardiomyocytes.

Mutations in human ether-a-go-go-related gene 1 (hERG) are linked to long QT syndrome type 2 (LQT2). hERG encodes the pore-forming alpha-subunits that coassemble to form rapidly activating delayed rectifier K(+) current in the heart. LQT2-linked missense mutations have been extensively studied in noncardiac heterologous expression systems, where biogenic (protein trafficking) and biophysical (gating and permeation) abnormalities have been postulated to underlie the loss-of-function phenotype associated with LQT2 channels. Little is known about the properties of LQT2-linked hERG channel proteins in native cardiomyocyte systems. In this study, we expressed wild-type (WT) hERG and three LQT2-linked mutations in neonatal mouse cardiomyocytes and studied their electrophysiological and biochemical properties. Compared with WT hERG channels, the LQT2 missense mutations G601S and N470D hERG exhibited altered protein trafficking and underwent pharmacological correction, and N470D hERG channels gated at more negative voltages. The DeltaY475 hERG deletion mutation trafficked similar to WT hERG channels, gated at more negative voltages, and had rapid deactivation kinetics, and these properties were confirmed in both neonatal mouse cardiomyocyte and human embryonic kidney (HEK)-293 cell expression systems. Differences between the cardiomyocytes and HEK-293 cell expression systems were that hERG current densities were reduced 10-fold and deactivation kinetics were accelerated 1.5- to 2-fold in neonatal mouse cardiomyocytes. An important finding of this work is that pharmacological correction of trafficking-deficient LQT2 mutations, as a potential innovative approach to therapy, is possible in native cardiac tissue.

Rescue of mutated cardiac ion channels in inherited arrhythmia syndromes.

Inherited arrhythmia syndromes comprise an increasingly complex group of diseases involving mutations in multiple genes encoding ion channels, ion channel accessory subunits and channel interacting proteins, and various regulatory elements. These mutations serve to disrupt normal electrophysiology in the heart, leading to increased arrhythmogenic risk and death. These diseases have added impact as they often affect young people, sometimes without warning. Although originally thought to alter ion channel function, it is now increasingly recognized that mutations may alter ion channel protein and messenger RNA processing, to reduce the number of channels reaching the surface membrane. For many of these mutations, it is also known that several interventions may restore protein processing of mutant channels to increase their surface membrane expression toward normal. In this article, we reviewed inherited arrhythmia syndromes, focusing on long QT syndrome type 2, and discuss the complex biology of ion channel trafficking and pharmacological rescue of disease-causing mutant channels. Pharmacological rescue of misprocessed mutant channel proteins, or their transcripts providing appropriate small molecule drugs can be developed, has the potential for novel clinical therapies in some patients with inherited arrhythmia syndromes.

The impact of asynchronous electroacupuncture stimulation duration on cold thermal pain threshold.

The durations of asynchronous electroacupuncture can affect the resultant hypoalgesia. Healthy volunteers were randomized to receive different durations (0 min, 20 min, 30 min, or 40 min) of asynchronous electroacupuncture stimulations (alternating low/high [2/100 Hz] frequency at 5 mA). Using a human experimental cold thermal pain threshold model, we found that 30 min of asynchronous 2/100 Hz stimulation resulted in the most significant hypoalgesic effect that was sustained for at least 60 min after stimulation compared with 0-, 20-, or 40-min stimulations (P < 0.05). We conclude that the most optimal duration for asynchronous electroacupuncture stimulation is 30 min.

Auricular acupuncture as a treatment for pregnant women who have low back and posterior pelvic pain: a pilot study.

OBJECTIVE: The primary aim of this study was to examine whether 1 week of continuous auricular acupuncture could reduce low back and posterior pelvic pain associated with pregnancy. STUDY DESIGN: A randomized controlled trial was conducted on pregnant women who have lower back and posterior pelvic pain. These women were randomly assigned into an acupuncture group, a sham acupuncture group, or a waiting list control group. All participants were monitored for 2 weeks. RESULTS: Baseline and day 7 showed significant group differences in pain (F = 15; P < .0001) and in the disability rating index score (F = 7; P < .0001). The participants in the acupuncture group reported a significant reduction of pain and improvement of functional status as compared with those in the sham acupuncture and control groups. CONCLUSION: One week of continuous auricular acupuncture decreases the pain and disability experienced by women with pregnancy-related low back and posterior pelvic pain.

Extra-1 acupressure for children undergoing anesthesia.

BACKGROUND: Acupuncture and related techniques have been used as adjuncts for perioperative anesthesia management. We examined whether acupressure in the Extra-1 (Yin-Tang) point would result in decreased preprocedural anxiety and reduced intraprocedural propofol requirements in a group of children undergoing endoscopic procedures. METHODS: Fifty-two children were randomized to receive acupressure bead intervention either at the Extra-1 acupuncture point or at a sham point. A Bispectral Index (BIS) monitor was applied to all children before the onset of the intervention. Anxiety was assessed at baseline and before entrance to the operating room. Anesthetic techniques were standardized and maintained with IV propofol infusion titrated to keep BIS values of 40-60. RESULTS: We found that after the intervention, children in the Extra-1 group experienced reduced anxiety whereas children in the sham group experienced increased anxiety (-9% [-3 to -15] vs 2% [-6 to 7.4], P = 0.012). In contrast, no significant changes in BIS values were observed in the preprocedural waiting period between groups (P = ns). We also found that total intraprocedural propofol requirements did not differ between the two study groups (214 +/- 76 microg x kg(-1) x min(-1) vs 229 +/- 95 microg x kg(-1) x min(-1), P = 0.52). CONCLUSIONS: We conclude that acupressure bead intervention at Extra-1 acupoint reduces preprocedural anxiety in children undergoing endoscopic procedures. This intervention, however, has no impact on BIS values or intraprocedural propofol requirements.