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.
阴极经颅直流电刺激(TDCS)诱导的耐药性癫痫发作的抑制作用。要进行有效的行动,刺激参数( 例如 ,方向,电场强度和持续时间的刺激),需要在老鼠大脑切片筹备工作进行检查。测试和安排相对于小鼠脑切片的位置的电极的取向是可行的。本方法保留thalamocingulate途径来评估DCS对前扣带皮层癫痫样活动的影响。多信道阵列记录的结果表明,阴极DCS显著降低的刺激诱发反应的4-氨基吡啶和荷包牡丹碱诱导的癫痫样活动的振幅和持续时间。这项研究还发现,在15分钟阴极DCS应用程序引起的长期萧条的thalamocingulate途径。本研究探讨DCS对thalamocingulat的影响Ë突触可塑性和急性发作之类的活动。当前程序可以测试最佳刺激参数,包括方向,磁场强度,并在体外的小鼠模型刺激持续时间。此外,该方法可以评估的DCS上皮层癫痫样活动在细胞和网络两级的作用。
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).
在本研究中,DCS系统上ACC癫痫样活动的持续时间和方向的影响进行了试验。以获得在小鼠脑切片稳定的数据,如何保持在MT-ACC通路的完整性,并避免损坏它是关键,特别是在其中两个成角度的腹侧削减和皮质的背切制成的步骤。此外,以制备脑切片的时间也可以影响脑切片,这应该是尽可能短的时间,以保持大脑新鲜和强的活性。先前的研究表明,对靶组织的电化学损伤可在体内制备15…
The authors have nothing to disclose.
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.
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 |