Fare Beyin Dilim Hazırlık içinde nöbet benzeri Faaliyet doğrudan akım Stimülasyon ve Multi-elektrot Dizi Kaydı
Summary
Studies have shown that cathodal transcranial direct-current stimulation can produce suppressive effects on drug-resistant seizures. In this study, an in vitro experimental setup was devised in which the direct-current stimulation and multielectrode array recording of seizure-like activity were evaluated in mice brain slice preparation. The direct-current stimulation parameters were evaluated.
Abstract
Katot transkranial doğru akım stimülasyonu (TEKMER) ilaca dirençli nöbetler üzerinde baskılayıcı etkisi neden olur. Etkili eylemleri gerçekleştirmek için, stimülasyon parametreleri (örneğin, yönlendirme, alan şiddeti ve stimülasyon süresi) fareler beyin dilim hazırlıkları muayene edilmesi gerekir. Test ve fareler beyin dilim konumuna elektrot göreceli yönünü düzenlenmesi mümkün bulunmaktadır. Mevcut yöntem anterior singulat korteks nöbet benzeri faaliyetleri DCS etkisini değerlendirmek için thalamocingulate yolu korur. Çok kanallı dizi kayıtların sonuçları katot DCS anlamlı 4-aminopiridin ve bicuculline kaynaklı nöbet benzeri aktivite uyarılması-uyarılmış yanıtlar genlik ve süresini azalttığını göstermiştir. Bu çalışma, 15 dakika ile katot DCS uygulamaları thalamocingulate yolunda uzun süreli depresyon neden olduğu bulunmuştur. Bu çalışma thalamocingulat üzerinde DCS etkilerini incelemektedirE sinaptik plastisite ve akut nöbet benzeri aktiviteler. Geçerli yordam bir in vitro fare modelinde yönlendirme, alan kuvveti ve stimülasyon süresi de dahil olmak üzere optimum stimülasyon parametreleri test edebilirsiniz. Ayrıca, yöntem hem hücresel hem de ağ seviyelerinde kortikal nöbet benzeri faaliyetleri DCS etkilerini değerlendirebilir.
Introduction
Epilepsy is a common neurological disorder. Thirty percent of patients with epilepsy suffer from drug-resistant seizures1. Transcranial direct-current stimulation (tDCS) provides a noninvasive approach to control or alter network activities across large brain areas, such as seizures. Clinical studies have shown that tDCS effectively treats intractable seizures2 and can produce both short- and long-term suppressive effects on seizures3-5. However, the therapeutic mechanism of tDCS actions is still unclear. The brain slice model presented is an in vitro method to investigate how the therapeutic mechanism of tDCS actions alters the symptoms of seizure-like brain activities. Accordingly, to achieve its optimal effects, specific stimulation parameters including orientation, field strength, and stimulation duration need to be tested in an experimental model. Previous studies have shown that the orientation of the electric field is important to obtain therapeutic effects6. Thus, testing and arranging the orientation of electrodes relative to the position of the tested brain slice are feasible.
Frontal lobe epilepsy and anterior cingulate cortex (ACC) seizures are often drug-resistant7,8. Some studies have reported the application of tDCS in the cingulate cortex9-11. tDCS is shown to affect vigilance, decision making and emotion through alteration of ACC activities, and can modulate neuronal excitability and seizure activity in this brain region12. Therefore, suppressive effects of tDCS on ACC seizures might be helpful for clinical treatment and the evaluation of alternative treatments.
The present protocol describes the preparation of an electrode in the recording chamber for DCS of a brain slice and its effect on seizure-like activity recording with a multielectrode array (MEA).
Protocol
Representative Results
Discussion
Bu çalışmada, ACC nöbet benzeri aktivite üzerine DCS süresine ve yönlendirme etkileri test edildi. fare beyin dilimleri kararlı veri elde etmek, nasıl MT-ACC yolunun bütünlüğünü korumak için ve iki açılı ventral kesikler ve korteksin dorsal kesim yapıldığı özellikle adımlar anahtarıdır zarar görmemesi için. Ayrıca, beyin dilim hazırlamak için zaman taze ve güçlü beyin tutmak için mümkün olan en kısa zaman olmalıdır beyin dilim, aktivitesini etkileyebilir. Önceki bir çalışmada…
Disclosures
The authors have nothing to disclose.
Acknowledgements
We are grateful for the technical support from the Neural Circuit Electrophysiology Core at Academia Sinica. This work was supported by the National Science Council (102-2320-B-001-026-MY3 and 100-2311-B-001-003-MY3) and Neuroscience Program of Academia Sinica.
Materials
| Anesthetic: | |||
| Isoflurane | Halocarbon Products Corporation | NDC 12164-002-25 | 4% |
| Name | Company | Catalog Number | Comments |
| aCSF (total:1L): | |||
| D(+)-Glucose | MERCK | 1.08337.1000 | 10 mM |
| Sodium hydrogen carbonate | MERCK | 1.06329.0500 | 25 mM |
| Sodium chloride | MERCK | 1.06404.1000 | 124 mM |
| (+)-Sodium L-ascorbate, >=98% | SIGMA | A4034-100G | 0.15 g / 2 c.c |
| Magnesium sulfate, anhydrous,ReagentPlus | SIGMA | M7506-500G | 2 mM |
| Calcium chloride dihydrate | MERCK | 1.02382.1000 | 2 mM |
| Sodium dihydrogen phosphate monohydrate | MERCK | 1.06346.1000 | 1 mM |
| Potassium chloride | May & Baker LTD Dagenham England | MS 7616 | 4.4 mM |
| Name | Company | Catalog Number | Comments |
| Drugs: | |||
| (+)-Bicuculline | TOCRIS | 0130 | 5 µM in aCSF |
| 4-Aminopyridine | TOCRIS | 0940 | 250 µM in aCSF |
| Name | Company | Catalog Number | Comments |
| Brain slice Preparation: | |||
| Vibratome | Vibratome | Series 1000 | Block slicing into 500 µm thick slices |
| Name | Company | Catalog Number | Comments |
| MEA system: | |||
| Multielectrode array (MEA) probes: 6 x 10 planar MEA | Multi Channel Systems | 60MEA500/30iR-Ti-pr MEAS 6×10 | electrode diameter, 30 µm; electrode spacing, 500 µm; impedance, 50 kΩ at 200 Hz |
| Multielectrode array (MEA) probes: 8 x 8 MEA | Ayanda Biosystems | 60MEA200/10iR-Ti-pr MEAS 8×8 | pyramidal-shaped electrode; diameter, 40 µm; tip height, 50 µm; electrode spacing, 200 µm; impedance, 1000 kΩ at 200 Hz |
| A 60-channel amplifier was used with a band-pass filter set between 0.1 Hz and 3 KHz at 1200X amplification | Multi-Channel Systems | MEA-1060-BC | |
| MC Rack software at a 10 KHz sampling rate | Multi-Channel Systems | Software for data collect and recordings | |
| control of a pulse generator | Multi-Channel Systems | STG 1002 | |
| slice anchor kits and hold-downs | Warner Instruments | SHD-26H/10; WI64-0250 | |
| Peristaltic Pump-minipuls3 | Gilsom | MINIPULS3 | perfusion rate : 8 ml/min |
| Name | Company | Catalog Number | Comments |
| Stimulation system: | |||
| Isolated stimulator | A-M Systems | Model 2100 | intensity of ±350 μA , duration of 200 μs |
| Tungsten electrode | A-M Systems | 575300 | placed in thalamus |
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