גירוי ישיר שוטף הקלטת מערך רבה-אלקטרודה של פעילות כמו-תפיסת עכברי הכנת פרוסת המוח
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 גירוי ישיר הנוכחי (tDCS) גורם תופעות מדכאות על התקפים עמידים לתרופות. כדי לבצע פעולות אפקטיביות, פרמטרי הגירוי (למשל, נטייה, עוצמת שדה, ומשך גירוי) צריכות להיבחן בהכנות פרוסות עכברי מוח. בדיקה וסידור שמיקומו ביחס האלקטרודה אל המיקום של פרוסת מוח עכברים הם ריאליות. השיטה הנוכחית שומרת על מסלול thalamocingulate כדי להעריך את ההשפעה של DCS על פעילויות, כמו תפיסת cingulate קורטקס. התוצאות של הקלטות מערך רב ציינו כי cathodal DCS משמעותית את המשרעת של עורריו תגובות הגירוי ומשך 4-aminopyridine ו bicuculline מושרה הפעילות כמו אפילפסיה. גם במחקר זה נמצא כי יישומי DCS cathodal ב 15 דק 'גורמים לדיכאון לטווח ארוך במסלול thalamocingulate. המחקר הנוכחי בוחן את ההשפעות של DCS על thalamocingulatדואר הפלסטיות הסינפטית ופעילויות, כמו תפיסת הכדור פנימה. ההליך הנוכחי יכול לבחון את פרמטרי גירוי האופטימליים כולל התמצאות, עוצמת שדה, ומשך גירוי במודל עכבר במבחנה. כמו כן, השיטה יכולה להעריך את ההשפעות של DCS על פעילויות קליפת מוח, כמו תפיסה בשתי רמות הסלולר ורשת.
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
במחקר הנוכחי, את ההשפעות של המשך והכיוון של DCS על פעילות התקפים דמויים ACC נבדקו. כדי לקבל נתונים יציבים פרוסות העכבר המוח, כיצד לשמור על שלמות של מסלול MT-ACC וכדי למנוע נזק זה הוא המפתח, במיוחד את הצעדים בהם שני חתכים הגחון זוויתי חתך הגבי של קליפת מבוצעים. יתר על כן, את הז?…
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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