Apigenin Alleviates Autistic-like Stereotyped Repetitive Behaviors and Mitigates Brain Oxidative Stress in Mice.

Studying the involvement of nicotinic acetylcholine receptors (nAChRs), specifically α7-nAChRs, in neuropsychiatric brain disorders such as autism spectrum disorder (ASD) has gained a growing interest. The flavonoid apigenin (APG) has been confirmed in its pharmacological action as a positive allosteric modulator of α7-nAChRs. However, there is no research describing the pharmacological potential of APG in ASD. The aim of this study was to evaluate the effects of the subchronic systemic treatment of APG (10-30 mg/kg) on ASD-like repetitive and compulsive-like behaviors and oxidative stress status in the hippocampus and cerebellum in BTBR mice, utilizing the reference drug aripiprazole (ARP, 1 mg/kg, i.p.). BTBR mice pretreated with APG (20 mg/kg) or ARP (1 mg/g, i.p.) displayed significant improvements in the marble-burying test (MBT), cotton-shredding test (CST), and self-grooming test (SGT) (all p < 0.05). However, a lower dose of APG (10 mg/kg, i.p.) failed to modulate behaviors in the MBT or SGT, but significantly attenuated the increased shredding behaviors in the CST of tested mice. Moreover, APG (10-30 mg/kg, i.p.) and ARP (1 mg/kg) moderated the disturbed levels of oxidative stress by mitigating the levels of catalase (CAT) and superoxide dismutase (SOD) in the hippocampus and cerebellum of treated BTBR mice. In patch clamp studies in hippocampal slices, the potency of choline (a selective agonist of α7-nAChRs) in activating fast inward currents was significantly potentiated following incubation with APG. Moreover, APG markedly potentiated the choline-induced enhancement of spontaneous inhibitory postsynaptic currents. The observed results propose the potential therapeutic use of APG in the management of ASD. However, further preclinical investigations in additional models and different rodent species are still needed to confirm the potential relevance of the therapeutic use of APG in ASD.

Effect of music rhythm magnetic field on long-term potentiation of hippocampal Schaffer-CA1 synapse plasticity.

Music and magnetic fields both play important regulatory roles in brain learning and memory. The present study aimed to investigate the effects of music rhythmic magnetic fields at different frequencies on long-term potentiation (LTP) in the Schaffer-CA1 region of the hippocampus, with the goal of elucidating the molecular mechanisms underlying the impact of music rhythmic magnetic fields on brain learning and memory. Three different frequency music tracks were selected, including soothing track 1: Courante from the Baroque Suite, medium-frequency track 2: saxophone version of Liang Zhu, and high-frequency track 3: Johann Pachelbel's music track Canon (trumpet version). Using an external sound card, power amplifier, and homemade coils, a time-varying magnetic field with a 2-mT music rhythm was produced to assess the effects of this magnetic field on LTP in the Schaffer-CA1 synapses of isolated rat hippocampal brain slices. The experimental results demonstrated that as the music frequency increased, the enhancing effect of the music rhythmic magnetic field on hippocampal synaptic plasticity LTP gradually intensified. Thus, high-frequency music rhythmic magnetic fields may offer a more effective means of enhancing LTP.

Regulation of LTP at rat hippocampal Schaffer-CA1 in vitro by musical rhythmic magnetic fields generated by red-pink (soothing) music tracks.

PURPOSE: Music therapy, like red-pink (soothing) music, is an important treatment for neurological disorders associated with learning and memory. Magnetic fields have been proved to have a similar regulating effect. However, the effect of magnetic fields with musical rhythm generated by the combination of the two has not been confirmed. This study aimed to investigate the regulation of magnetic stimulation with music rhythm on LTP (long-term potentiation) of Schaffer-CA1. MATERIALS AND METHODS: This article selected three sorts of music tracks in different frequencies (music track (1) Turkish March, music track (2) Moonlight Sonata, music track (3) Funeral March) and four sorts of pure sinusoidal tracks of four different harmonic frequency (music track (4) the frequency is 3500 Hz; music track (5) the frequency is 2500 Hz; music track (6) the frequency is 1500 Hz; music track (7) the frequency is 500 Hz). These music tracks are converted into analog signals by the external sound card and power amplifier and fed into a homemade coil that meets the demand for this frequency bandwidth. The coil can generate seven sorts of time-varying magnetic fields with musical rhythm with a mean intensity of about 2 mT. We used multi-electrode array (MEA) to record the LTP signals of Schaffer-CA1 synaptic induced by seven sorts of musical rhythmic magnetic fields and analyze the regulation of them. RESULTS: The musical rhythmic magnetic fields generated by track 1 and track 2 have a remarkable enhancing effect on the amplitude of fEPSPs (field excitatory postsynaptic potentials) (p < .05), and these effects intensify with the increase of frequency. Nevertheless, there is no significant enhancing effect on LTP of the rhythmic magnetic field generated by track 3 (p > .05). The sinusoidal magnetic fields generated by track 4 and track 5 have an enhancing effect on the amplitude of fEPSPs (p < .05), and the enhancement is better than track 1 and track 2. The sinusoidal magnetic fields generated by track 6 and track 7 have an inhibiting effect (p < .05). CONCLUSION: We found that the enhancing effect of musical rhythmic magnetic fields generated by track 1 was the most significant. The frequency of 1500 Hz could be a turning-point frequency in the regulation of magnetic field on LTP.

A 6-Step Approach to Gain Higher Quality Results From Organotypic Hippocampal Brain Slices in a Traumatic Brain Injury Model.

Introduction: Organotypic Hippocampal Brain Slices (OHBS) provide an advantageous alternative to in vivo models to scrutinize Traumatic Brain Injury (TBI). We followed a well-established TBI protocol, but noticed that several factors may influence the results in such a setup. Here, we describe a structured approach to generate more comparable results and discuss why specific eligibility criteria should be applied. Methods: We defined necessary checkpoints and developed inclusion and exclusion criteria that take the observed variation in such a model into consideration. Objective measures include the identification and exclusion of pre-damaged slices and outliers. Six steps were outlined in this study. Results: A six-step approach to enhance comparability is proposed and summarized in a flowchart. We applied the suggested measures to data derived from our TBI-experiments examining the impact of three different interventions in 1459 OHBS. Our exemplary results show that through equal requirements set for all slices more precise findings are ensured. Conclusion: Results in a TBI experiment on OHBS should be analyzed critically as inhomogeneities may occur. In order to ensure more precise findings, a structured approach of comparing the results should be followed. Further research is recommended to confirm and further develop this framework.

Arctic ground squirrel resist peroxynitrite-mediated cell death in response to oxygen glucose deprivation.

Cerebral ischemia-reperfusion (I/R) injury initiates a cascade of events, generating nitric oxide (NO) and superoxide(O2•-) to form peroxynitrite (ONOO-), a potent oxidant. Arctic ground squirrels (AGS; Urocitellus parryii) show high tolerance to I/R injury. However, the underlying mechanism remains elusive. We hypothesize that tolerance to I/R modeled in an acute hippocampal slice preparation in AGS is modulated by reduced oxidative and nitrative stress. Hippocampal slices (400µm) from rat and AGS were subjected to oxygen glucose deprivation (OGD) using a novel microperfusion technique. Slices were exposed to NO, O2.- donors with and without OGD; pretreatment with inhibitors of NO, O2.- and ONOO- followed by OGD. Perfusates collected every 15min were analyzed for LDH release, a marker of cell death. 3-nitrotyrosine (3NT) and 4-hydroxynonenal (4HNE) were measured to assess oxidative and nitrative stress. Results show that NO/O2.- alone is not sufficient to cause ischemic-like cell death, but with OGD enhances cell death more in rat than in AGS. A NOS inhibitor, SOD mimetic and ONOO- inhibitor attenuates OGD injury in rat but has no effect in AGS. Rats also show a higher level of 3NT and 4HNE with OGD than AGS suggesting the greater level of injury in rat is via formation of ONOO-.

Argon attenuates the emergence of secondary injury after traumatic brain injury within a 2-hour incubation period compared to desflurane: an in vitro study.

Despite years of research, treatment of traumatic brain injury (TBI) remains challenging. Considerable data exists that some volatile anesthetics might be neuroprotective. However, several studies have also revealed a rather neurotoxic profile of anesthetics. In this study, we investigated the effects of argon 50%, desflurane 6% and their combination in an in vitro TBI model with incubation times similar to narcotic time slots in a daily clinical routine. Organotypic hippocampal brain slices of 5- to 7-day-old mice were cultivated for 14 days before TBI was performed. Slices were eventually incubated for 2 hours in an atmosphere containing no anesthetic gas, argon 50% or desflurane 6% or both. Trauma intensity was evaluated via fluorescent imagery. Our results show that neither argon 50% nor desflurane 6% nor their combination could significantly reduce the trauma intensity in comparison to the standard atmosphere. However, in comparison to desflurane 6%, argon 50% displayed a rather neuroprotective profile within the first 2 hours after a focal mechanical trauma (P = 0.015). A 2-hour incubation in an atmosphere containing both gases, argon 50% and desflurane 6%, did not result in significant effects in comparison to the argon 50% group or the desflurane 6% group. Our findings demonstrate that within a 2-hour incubation time neither argon nor desflurane could affect propidium iodide-detectable cell death in an in vitro TBI model in comparison to the standard atmosphere, although cell death was less with argon 50% than with desflurane 6%. The results show that within this short time period processes concerning the development of secondary injury are already taking place and may be manipulated by argon.

Arctic ground squirrel hippocampus tolerates oxygen glucose deprivation independent of hibernation season even when not hibernating and after ATP depletion, acidosis, and glutamate efflux.

Cerebral ischemia/reperfusion (I/R) triggers a cascade of uncontrolled cellular processes that perturb cell homeostasis. The arctic ground squirrel (AGS), a seasonal hibernator resists brain damage following cerebral I/R caused by cardiac arrest and resuscitation. However, it remains unclear if tolerance to I/R injury in AGS depends on the hibernation season. Moreover, it is also not clear if events such as depletion of ATP, acidosis, and glutamate efflux that are associated with anoxic depolarization are attenuated in AGS. Here, we employ a novel microperfusion technique to test the hypothesis that tolerance to I/R injury modeled in an acute hippocampal slice preparation in AGS is independent of the hibernation season and persists even after glutamate efflux. Acute hippocampal slices were harvested from summer euthermic AGS, hibernating AGS, and interbout euthermic AGS. Slices were subjected to oxygen glucose deprivation (OGD), an in vitro model of I/R injury to determine cell death marked by lactate dehydrogenase (LDH) release. ATP was assayed using ENLITEN ATP assay. Glutamate and aspartate efflux was measured using capillary electrophoresis. For acidosis, slices were subjected to pH 6.4 or ischemic shift solution (ISS). Acute hippocampal slices from rats were used as a positive control, susceptible to I/R injury. Our results indicate that when tissue temperature is maintained at 36°C, hibernation season has no influence on OGD-induced cell death in AGS hippocampal slices. Our data also show that tolerance to OGD in AGS hippocampal slices occurs despite loss of ATP and glutamate release, and persists during conditions that mimic acidosis and ionic shifts, characteristic of cerebral I/R. Read the Editorial Comment for this article on page 10.