चूहे मस्तिष्क टुकड़ा तैयारी में जब्ती की तरह गतिविधि के प्रत्यक्ष वर्तमान उत्तेजना और मल्टी-इलेक्ट्रोड सरणी रिकॉर्डिंग
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
Cathodal Transcranial प्रत्यक्ष वर्तमान उत्तेजना (tDCS) दवा प्रतिरोधी दौरे पर दमनकारी प्रभाव लाती है। प्रभावी कार्रवाई करने के लिए, उत्तेजना मानकों (जैसे, ओरिएंटेशन, क्षेत्र की ताकत है, और उत्तेजना की अवधि) चूहों के मस्तिष्क टुकड़ा तैयारी में जांच की जानी चाहिए। परीक्षण और चूहों के मस्तिष्क टुकड़ा की स्थिति के लिए इलेक्ट्रोड रिश्तेदार के उन्मुखीकरण की व्यवस्था संभव है। वर्तमान पद्धति पूर्वकाल सिंगुलेट प्रांतस्था जब्ती-तरह की गतिविधियों पर डीसीएस के प्रभाव का मूल्यांकन करने के लिए thalamocingulate मार्ग को बरकरार रखता है। मल्टीचैनल सरणी रिकॉर्डिंग के परिणाम संकेत दिया कि cathodal डीसीएस काफी उत्तेजना पैदा प्रतिक्रियाओं के आयाम और 4-aminopyridine और bicuculline प्रेरित जब्ती की तरह गतिविधि की अवधि में कमी आई है। इस अध्ययन में यह भी पाया गया है कि 15 मिनट पर cathodal डीसीएस अनुप्रयोगों thalamocingulate मार्ग में लंबे समय तक अवसाद का कारण बना। वर्तमान अध्ययन thalamocingulat पर डीसीएस के प्रभावों की जांचई synaptic plasticity और तीव्र जब्ती-तरह की गतिविधियों। वर्तमान प्रक्रिया अभिविन्यास, क्षेत्र की ताकत है, और इन विट्रो माउस मॉडल में उत्तेजना अवधि सहित इष्टतम उत्तेजना मापदंडों का परीक्षण कर सकते हैं। इसके अलावा, विधि दोनों सेलुलर और नेटवर्क के स्तर पर cortical जब्ती-तरह की गतिविधियों पर डीसीएस के प्रभाव का मूल्यांकन कर सकते हैं।
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
वर्तमान अध्ययन में, एसीसी जब्ती की तरह गतिविधि पर अवधि और डीसीएस के उन्मुखीकरण के प्रभाव का परीक्षण किया गया। माउस मस्तिष्क स्लाइस में स्थिर डेटा प्राप्त करने के लिए, कैसे मीट्रिक टन एसीसी मार्ग की अख?…
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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