Name: simultaneous electrophysiological recording and micro-injections of inhibitory agents in the rodent brain
Uploaded: 2015-07-07T13:00:00.000+00:00
Duration: 7 min 52 s
Description: Watch this Scientific Journal Video about Simultaneous Electrophysiological Recording and Micro-injections of Inhibitory Agents in the Rodent Brain at JoVE.com

Supported by grants from CIHR (MOP231122) and NSERC (RGPIN-2014-06503). We would like to thank Genevi&#232;ve Cyr for her help preparing experiments and supervising laboratory work. MAL received a scholarship from The Natural Sciences and Engineering Research Council of Canada (NSERC).

참고 : 모든 절차는 동물의 보호와 몬트리올 대학교의 윤리 검토위원회의 캐나다 협의회의 지침에 따라 수행되었다. 녹화 주입 피펫 1. 총회 <ol><li> 피펫 풀러를 사용하여 약 7cm 긴 유리 모세관 (1mm 외경)을 당깁니다. </li><li> 모세관의 끝을 휴식과 가벼운 현미경 조리개를 확인합니다. 내부 직경이 40 μm의 30 μm의 사이에 있는지 확인합니다. </li><li> 유리 피펫의 비 테이퍼 끝에서 돌출 약 1cm와 유리 모세관에 7cm 길이 실버 와이어를 삽입합니다. </li><li> 유리 모세관에 직교 초과 필라멘트를 구부리고. </li><li> 30 G 피하 주사 바늘의 축에 유연한 플라스틱 접착제의 방울을 적용합니다. </li><li> 그림 1에서 제시 한 설계도에 따라 유리 피펫에 30 G 피하 주사 바늘을 ​​삽입 </stro&gt; NG. </li><li> 유리 피펫 및 피하 주사 바늘 사이의 접합에서 적절한 밀봉을 보장하기 위해 접착제의 두 번째 코팅을 추가합니다. </li><li> 끝이 접착제의 적절한 경화를 보장하기 위해 약 12​​ 시간 동안 위로 향과 건조 피펫을 남겨주세요. 최종 결과는 그림 2에 표시됩니다. </li></ol> 참고 :이 절차는 급성 실험과 피펫 팁의 살균에 수행 할 필요가 없습니다. 2. 동물 준비 <ol><li> 마취 상자에 쥐를 놓습니다. </li><li> 5 내지 10 분 동안 4 % 이소 플루 란을 이용하여 마취를 유도한다. </li><li> 37 ° C의 체온을 유지하기 위해 가열 패드 및 직장 프로브 정위 테이블에 동물을 놓는다. 2 % 이소 플루 란 마취를 유지하기 위해 코 원뿔을 사용한다. 귀 바, 치아 홀더를 사용하여 쥐의 머리를 고정합니다. </li><li> 아트로핀은 동공 팽창에 도움이 떨어진다 1 %를 포함 안과 연고 또는 눈 방울을 적용합니다. 건조 함을 방지하기 위해, 드롭 윤활 적용약 30 분마다이야. </li><li> 머리를 면도 10 % 포비돈 - 요오드로 청소. </li><li> 국소 마취를 들면, 피부를 리프팅하고 바늘의 선단부를 삽입하여 위치 2-3에서 두피 아래에 2 % 리도카인 0.5 ml를 주입. </li><li> 발가락 핀치를 수행하고 운동의 부족을 관찰하여 마취의 적절한 수준을 확인합니다. 또한, 정상 값 (300-400 비트 / 분) 이내인지 확인 심박수를 모니터링. </li><li> 관상 및 시상 봉합을 모두 노출 # 10 메스 블레이드와 중간을 따라 직선으로 두피를 절개. </li><li> 수술 주걱으로 두개골을 덮고있는 조직을 옆으로 밀어 람다와 브레 그마 점을 알 수있다. </li><li> 브레 그마 및 람다 위치가 동일 평면에 있도록 수평을 두개골. </li><li> 참조 포인트를 설정하기 위해, 바로 위의 브레 그마로 설정하도록 장착 된 유리 튜브로 정위 장치를 사용한다. 이것은 &quot;제로&quot;에 대한 안테 - 후방 및 의대 될 것입니다IAL-측면 측정을 조정합니다. </li><li> , 정위 필요한 좌표로 마운트 이동하여 관심 영역을 설정 정위 좌표를 기록하고 개두술이 수행 될 위치를 표시하는 대상 영역 주위에 사각형을 그립니다. </li><li> 천천히 뼈 물질을 제거하기 위해 압력없이 천천히 표시된 사각형 따라 멸균 드릴 수술 드릴을 사용합니다. 이 피질에 병변이 열을 발생 원인이되므로, 같은 지역에 너무 오래 드릴하지 않도록주의하십시오. </li><li> 개두술을 구분 뼈가 충분히 얇은 될 때,주의 깊게 피질을 노출 핀셋 두개골 부분을 제거합니다. </li><li> 자주 조직 탈수를 방지하기 위해 인공 뇌척수액에 노출 된 피질을 관개. 주 : injectrode의 끝이 침투 할만큼 튼튼한 같이 경막 제거가 쥐에 필요하지 않습니다. </li></ol> 3. 채우고 주입 시스템의 설치 <ol><li> 작성미네랄 오일과 흡인에 의해 5 ~ 10 μL를 마이크로 실린. </li><li> 0.5 % 시카고 스카이 블루 (CSB) 및 300 μM의 γ - 아미노 부티르산 (GABA) 또는 0.5 % CSB 9 2 % 리도카인 용액의 용액으로 고정 유리 피펫으로 피하 주사 바늘을 ​​입력합니다. 식염수 모든 솔루션을 희석. <ol><li> 풍부한 물질의 경우에, 주사기를 사용하여 정기적으로 채우고 기포의 형성을 피하기 위해 평소 예방 기법을 사용한다. </li><li> 더 비싼 물질의 경우, injectrode 채우기 위해 미네랄 오일을 사용하고 화학 제는 다음 흡인에 의해 도입 될 수있다. 미네랄 오일과 물 사이의 밀도 차이가 비교적 높고,이 물질은 수용액의 주입을위한 좋은 후보이다. 주 : 염료 모두 액체 사이의 거리를 확인하기 위해 추가 될 수있다. </li></ol></li><li> 주입 피펫을 채우려면, 흡인에 의한 용액 1 ML의 주사기를 기입 한 후 천천히 솔루션을 주입주입 피펫으로. </li><li> 깨끗한이 지역을 보라고 더욱 주사기와 천천히 용액을 주입하여 누출을 관찰하여 그림 1에 표시된 지역에서 누출 세심한주의를 기울이십시오. </li><li> 1 ML의 주사기를 제거합니다. 이렇게되면, 진공 주입 피펫의 용액을 제거하지 않도록 플런저에 가벼운 압력을 유지해야합니다. </li><li> 흡인에 의해 미네랄 오일과 마이크로 실린를 채우고 가득 주입 피펫에 단단히 부착, 조심스럽게 거즈와 조립 injectrode에서 초과 솔루션을 닦아. </li><li> 끝이 유리 피펫의 끝에서 형성 작은 ​​방울을 볼 수있을만큼 아주 작은 볼륨을 주입에 의해 차단되지 않았는지 확인합니다. </li><li> 마이크로 시스템의 injectrode를 탑재하고 잘 고정되어 있는지 확인합니다. </li><li> 조심스럽게 목표 좌표 injectrode 팁을 놓고 피질의 표면에 팁을 낮 춥니 다. </li><li> 천천히 injectr을 낮출적절한 해부 좌표를 사용하여 상기 타겟 구조 정위 장치 (이 경우에는 우수한 둔덕)를 사용 ODE. </li><li> 조직의 탈수를 방지하기 위해 따뜻한 한천에 노출 된 피질을 커버. </li></ol> 4. 사출 및 가역 불 활성화 <ol><li> 주입을 시작하는 40 NL / 분 키를 누릅니다 실행에 400 NL (800)에 주입하는 미세 주입 펌프를 설정합니다. 그 스파이크 속도가 주입 동안 감소를 보여줍니다 있습니다. 참고 : 실험 설정에서 신경 활동은 GABA 전달의 종료 후 시간 이내에 복구. 상관 조정 기계 장치는 분사를 실시하기 위해 마이크로 인젝션 주사기에 압력을 적용하는데 사용될 수있다. </li><li> 전기 생리 데이터를 수집 한 후, 지역의 동물 윤리 커뮤니티 승인 한 방법을 사용하여 안락사. </li></ol>

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We have nothing to disclose.

현재 제안 된 프로토콜은 가역적 불활 방법에서 발생하는 문제점을 해결하기 위해 설계되었다. 특히,이 프로젝트는 특히 깊은 뇌 구조에서, 신경 활동을 조절 물질의 화학 microinjections에 사용되는 방법을 구체화하기위한. 이러한 유형의 설정 신흥 기술 과제는 모두 프로브는 주사 부위에서 정확한 기록을 유도하기 위해 생체 내에서 같은 제한된 공간에서 colocalized하기 위해서는 필요하다. ?...

피질 영역과 서브 피질 핵의 불 활성화는 다양한 뇌 구조 2-4 사이의 기능적 관계의 연구에서 중요하다. 최근 문학 뇌 구조 2,5의 역할을 연구하는 기능 상실 화학 물질 또는 극저온 기술을 채용하고있다. 주변 조직에 -6,7- 담보 손상을 최소화하면서 약리학 microinjections에 관련하여, 약물의 작은 볼륨 조절 된 속도로 뇌 영역으로 투여 될 수있다. 이 기술은 신경 세포 활성도에 대한 다른 약물 표적의 효과를 연구하기 위해 특정한 효능 제, 역 효능 제 또는 길항제를 전달하는데 사용될 수있다. 이러한 효과는 또한 다른 연구자 피질과 피질 하 구조 사이의 관계를 연구 할 수 있도록 먼 위치에서 신경 반응의 변화를 측정함으로써 조사 될 수있다. 여기서는 보 수있는 장치의 조립, injectrode을 보여전기 생리 신호를 기록 및 목표 위치에서의 약물 전달을 소량 번째. 우리 래트에서 우수한 둔덕, GABA, 신경 세포 활성도의 일반적인 억제제를 주입하여이 시스템의 능력을 입증한다. 이 지역은 우리가 injectrode 현지화를 확인하기 위해 시각적으로 유발 멀티 유닛 활동을 사용할 수 시각적 자극에 민감하다. 불 활성화의 가역성은 GABA 분사 종료 후 정상적인 신경 활성의 회복에 의해 평가 하였다. 주사 부위에서 멀티 유닛 활동을 모니터링 할 수있는 능력은 바람직한 약물 동태 학적 반응을 달성하는데 필요한 사출 속도와 볼륨의 미세 조정을 허용한다. 최소 유효 부피가 주입되기 때문에, 따라서,이 기술의 장점은, microperfusion으로 인한 조직 손상을 제한하는 전위이다. 제안 된 프로토콜은 일회용 하드웨어 necessar를 생성하기위한 비용 효율적인 방법을 제공한다약물 전달과 지역의 연결 활동 기록이 요구되는 수행 실험 Y.

<table><tbody><tr><td>Injection pump (UltraMicroPump III)</td><td>WPI</td><td>#UMP3</td><td></td></tr><tr><td>Injection console (Micro4 Controller)</td><td>WPI</td><td>#SYS-MICRO4</td><td></td></tr><tr><td>Hamilton syringe</td><td>Hamliton</td><td>(80301) 701LT 10 &amp;micro;L SYR</td><td>Syringes between 5 and 10 &amp;mu;l used</td></tr><tr><td>Flexible plastic adhesive</td><td>Lepage</td><td>393915</td><td>The gel form is easier to apply to the shaft of the 30&amp;nbsp;G hypodermic needle forming a waterproof seal when dry.</td></tr><tr><td>Glass pipettes</td><td>WPI</td><td>#TW100F-4</td><td>Thin wall, 1&amp;nbsp;mm OD, 0.75&amp;nbsp;mm ID with filament pipettes used</td></tr><tr><td>720 Needle Pipette Puller</td><td>Kopf</td><td>720</td><td></td></tr><tr><td>Silver wire</td><td>A-M Systems, Inc.</td><td>782500</td><td>Bare 0.010&amp;rdquo;</td></tr></tbody></table>

simultaneous electrophysiological recording and micro-injections of inhibitory agents in the rodent brain

여기서 우리는 상업적으로 접근하고 저렴한 부품을 이용하여 단일 사용 &quot;injectrode&quot;의 건설 방법을 설명한다. 프로빙 시스템은 영향받은 뉴런 집단에서 전기 생리 신호를 기록하는 동안 약물의 주입을 허용하는 개발되었다. 이 방법은 상용 솔루션을 간단하고 경제적 인 대안을 제공합니다. 유리 피펫은 피하 주사 바늘과 실버 필라멘트와 결합하여 수정되었습니다. injectrode은 약물 전달을위한 상업 마이크로 실린 펌프에 연결되어 있습니다. 이것은 약물 전달의 사이트에서 유래 다세대 세포 외 신호를 통해 실시간 약력학 피드백을 제공하는 기술을 초래한다. 개념의 증거로서, 우리는 병용 injectrode을 통해 약물의 배달, 쥐에서 빛의 섬광에 의해 유발 우수한 둔덕에서 신경 세포의 활동을 기록했다. injectrode 기록 용량 INJ의 기능적 특성을 허용약물 전달의 현지화를 정확하게 제어 선호 ection 사이트. injectrode에로드 된 화학 물질의 선택은 해부학 실험 마커를 추적 포함, 광대로이 방법의 응용 프로그램은 또한, 지금까지 여기에 입증되는 것 이상으로 확장합니다.

injectrode의 구조는 와이어의 절곡 부와 유리 피펫 (D)로 공급되고, 개구로부터 밖으로 돌출되어있다도 1은 와이어 (C)에 도시되어있다. 30 G 바늘 (B)는 부착 접착제와 유리 피펫의 개통에 밀봉된다. 피펫은 주입 물질로 충전 된 후, 유리 마이크로 주사기 (A)가 바늘에 부착된다. 이것은 마이크로 주사기 바늘 (E) 및 여기서 은선은 유리 피펫 (F)로부터 돌출와 연결 양호한 시일이 존재하는 것...

Watch this Scientific Journal Video about Simultaneous Electrophysiological Recording and Micro-injections of Inhibitory Agents in the Rodent Brain at JoVE.com

Simultaneous Electrophysiological Recording and Micro-injections of Inhibitory Agents in the Rodent Brain

Here, we craft a glass pipette with dual functions: inhibition of deep brain structures by microinjections of drugs and real-time monitoring of their effects through simultaneous electrophysiological recordings.

동시 전기 생리 녹화 및 설치류의 뇌에서 억제 에이전트의 마이크로 주사

Here we describe a method for the construction of a single-use &#8220;injectrode&#8221; using commercially accessible and affordable parts. A probing ...

simultaneous-electrophysiological-recording-micro-injections

Research

JoVE Journal

Neuroscience

8.1K 조회수.  Université de Montréal. Here, we craft a glass pipette with dual functions: inhibition of deep brain structures by microinjections of drugs and real-time monitoring of their effects through simultaneous electrophysiological recordings.

비디오: Simultaneous Electrophysiological Recording and Micro-injections of Inhibitory Agents in the Rodent Brain

Simultaneous fMRI and Electrophysiology in the Rodent Brain

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in which functional MRI data and in vivo intracortical recording can be performed simultaneously. The combination of MRI and electrical recording is technically challenging because the electrodes used for recording distort the MRI images and the MRI acquisition induces noise in the electrical recording. To minimize the mutual interference of the two modalities, glass microelectrodes were used rather than metal and a noise removal algorithm was implemented for the electrophysiology data. In our studies, two microelectrodes were separately implanted in bilateral primary somatosensory cortices (SI) of the rat and fixed in place. One coronal slice covering the electrode tips was selected for functional MRI. Electrode shafts and fixation positions were not included in the image slice to avoid imaging artifacts. The removed scalp was replaced with toothpaste to reduce susceptibility mismatch and prevent Gibbs ringing artifacts in the images. The artifact structure induced in the electrical recordings by the rapidly-switching magnetic fields during image acquisition was characterized by averaging all cycles of scans for each run. The noise structure during imaging was then subtracted from original recordings. The denoised time courses were then used for further analysis in combination with the fMRI data. As an example, the simultaneous acquisition was used to determine the relationship between spontaneous fMRI BOLD signals and band-limited intracortical electrical activity. Simultaneous fMRI and electrophysiological recording in the rodent will provide a platform for many exciting applications in neuroscience in addition to elucidating the relationship between the fMRI BOLD signal and neuronal activity.

We have developed a method for simultaneous functional magnetic resonance imaging and electrophysiological recording in the rodent brain, providing a platform for the investigation of the relationship between neural activity and the blood oxygenation level dependent (BOLD) MRI signal.

쥐 두뇌에 동시 fMRI와 전기 생리학

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in ...

연구

신경과학

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

동시 Electroencephalography, 설치류 대뇌 피질의 젖산 농도와 Neuronal 활동 Optogenetic 조작의 실시간 측정

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and awake animals. Therefore, methods allowing the manipulation of synaptic systems in awake animals are required in order to determine the contribution of synaptic inputs to neuronal processing unaffected by anesthetics. Here, we present methodology for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. By combining these procedures, we performed microiontophoretic injections of gabazine to selectively block GABAA receptors in neurons of the inferior colliculus of head-restrained mice. Gabazine successfully modified neural response properties such as the frequency response area and stimulus-specific adaptation. Thus, we demonstrate that our methods are suitable for recording single-unit activity and for dissecting the role of specific neurotransmitter receptors in auditory processing.
The main limitation of the described procedure is the relatively short recording time (~3 hr), which is determined by the level of habituation of the animal to the recording sessions. On the other hand, multiple recording sessions can be performed in the same animal. The advantage of this technique over other experimental procedures used to manipulate the level of neurotransmission or neuromodulation (such as systemic injections or the use of optogenetic models), is that the drug effect is confined to the local synaptic inputs to the target neuron. In addition, the custom-manufacture of electrodes allows adjustment of specific parameters according to the neural structure and type of neuron of interest (such as the tip resistance for improving the signal-to-noise ratio of the recordings).

We present methods for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. This technique allows the detailed analysis of putative local synaptic inputs to the neuron of interest.

깨어 마우스의 신경 활성 물질의 Microiontophoretic 응용 프로그램과 결합하여 신경 활동의 세포 외 기록

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and ...

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the pathophysiological basis of these diseases, it is necessary to precisely characterize in which way the processing of information is disturbed between the different neuronal parts of the circuit. Using extracellular in vivo electrophysiological recordings, it is possible to accurately delineate neuronal activity within a neuronal network. The application of this method has several advantages over alternative techniques, e.g., functional magnetic resonance imaging and calcium imaging, as it allows a unique temporal and spatial resolution and does not rely on genetically engineered organisms. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied. In this article, a simple and easy to use method is presented with which it is possible to simultaneously record extracellular potentials such as local field potentials and multiunit activity at multiple sites of a network. It is detailed how a precise targeting of subcortical nuclei can be achieved using a combination of stereotactic surgery and online analysis of multi-unit recordings. Thus, it is demonstrated, how a complete network such as the hyperdirect cortico-basal ganglia loop can be studied in anesthetized animals in vivo.

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

심각한<em&gt; In Vivo</em&gt; 마취 된 래트에서의 Hyperdirect 경로로부터의 국소 장 전위 및 다중 단위 활성의 전기 생리 학적 기록

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the neurogenesis of EEG is still very limited. Local field potentials (LFPs) recorded via a multi-laminar microelectrode can provide a more detailed account of simultaneous neural activity across different cortical layers in the neocortex, but the technique is invasive. Combining EEG and LFP measurements in a pre-clinical model can greatly enhance understanding of the neural mechanisms involved in the generation of EEG signals, and facilitate the derivation of a more realistic and biologically accurate mathematical model of EEG. A simple procedure for acquiring concurrent and co-localized EEG and multi-laminar LFP signals in the anesthetized rodent is presented here. We also investigated whether EEG signals were significantly affected by a burr hole drilled in the skull for the insertion of a microelectrode. Our results suggest that the burr hole has a negligible impact on EEG recordings.

This protocol describes a simple method for concurrent recording of co-localized electroencephalography (EEG) and multi-laminar local field potential in an anesthetized rat. A burr hole drilled in the skull for the insertion of a microelectrode is shown to produce negligible distortion of the EEG signal.

공동 화 된 Electroencephalography 및 설치류에서 로컬 필드 가능성의 동시 녹음

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the ...

Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) is a promising approach when pharmacological treatment fails or neurosurgery is not recommended. Acute brain slices coupled to microelectrode arrays (MEAs) represent a valuable tool to study neuronal network interactions and their modulation by electrical stimulation. As compared to conventional extracellular recording techniques, they provide the added advantages of a greater number of observation points and a known inter-electrode distance, which allow studying the propagation path and speed of electrophysiological signals. However, tissue oxygenation may be greatly impaired during MEA recording, requiring a high perfusion rate, which comes at the cost of decreased signal-to-noise ratio and higher oscillations in the experimental temperature. Electrical stimulation further stresses the brain tissue, making it difficult to pursue prolonged recording/stimulation epochs. Moreover, electrical modulation of brain slice activity needs to target specific structures/pathways within the brain slice, requiring that electrode mapping be easily and quickly performed live during the experiment. Here, we illustrate how to perform the recording and electrical modulation of 4-aminopyridine (4AP)-induced epileptiform activity in rodent brain slices using planar MEAs. We show that the brain tissue obtained from mice outperforms rat brain tissue and is thus better suited for MEA experiments. This protocol guarantees the generation and maintenance of a stable epileptiform pattern that faithfully reproduces the electrophysiological features observed with conventional field potential recording, persists for several hours, and outlasts sustained electrical stimulation for prolonged epochs. Tissue viability throughout the experiment is achieved thanks to the use of a small-volume custom recording chamber allowing for laminar flow and quick solution exchange even at low (1 mL/min) perfusion rates. Quick MEA mapping for real-time monitoring and selection of stimulating electrodes is performed by a custom graphic user interface (GUI).

We illustrate how to perform recording and electrical modulation of 4-aminopyridine-induced epileptiform activity in rodent brain slices using microelectrode arrays. A custom recording chamber maintains tissue viability throughout prolonged experimental sessions. Live electrode mapping and selection of stimulating pairs are performed by a custom graphical user interface.

녹음 및 Microelectrode 배열에 결합 하는 설치류 뇌 조각에 Epileptiform 활동의 변조

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) ...

Microinjectrode System for Combined Drug Infusion and Electrophysiology

This microinjectrode system is designed for drug infusion, electrophysiology, and delivery and retrieval of experimental probes, such as microelectrodes and nanosensors, optimized for repeated use in awake, behaving animals. The microinjectrode system can be configured for multiple purposes: (1) simple arrangement of the cannula for placement of an experimental probe that would otherwise be too fragile to penetrate the dura mater, (2) microfluidic infusion of a drug, either independently or coupled to a cannula containing an experimental probe (i.e., microelectrode, nanosensor). In this protocol we explain the step by step construction of the microinjectrode, its coupling to microfluidic components, and the protocol for use of the system in vivo. The microfluidic components of this system allow for delivery of volumes on the nanoliter scale, with minimal penetration damage. Drug infusion can be performed independently or simultaneously with experimental probes such as microelectrodes or nanosensors in an awake, behaving animal. Applications of this system range from measuring the effects of a drug on cortical electrical activity and behavior, to understanding the function of a specific region of cortex in the context of behavioral performance based on probe or nanosensor measurements. To demonstrate some of the capabilities of this system, we present an example of muscimol infusion for reversible inactivation of the frontal eye field (FEF) in rhesus macaque during a working memory task.

We present a microinjectrode system designed for electrophysiology and assisted delivery of experimental probes (i.e., nanosensors, microelectrodes), with optional drug infusion. Widely available microfluidic components are coupled to a cannula containing the probe. A step-by-step protocol for microinjectrode construction is included, with results during muscimol infusion in macaque cortex.

결합 된 약물 주입 및 전기 생리학을위한 마이크로 인젝트 로드 시스템

This microinjectrode system is designed for drug infusion, electrophysiology, and delivery and retrieval of experimental probes, such as microelectrodes ...

Drug Microinjection with Electrophysiological Recording in a Rat Brain

Source: Lai, J., et al. <a href="https://app.jove.com/v/52271">Simultaneous electrophysiological recording and micro-injections of inhibitory agents in the rodent brain.</a> J. Vis. Exp. (2015)
This protocol outlines the assembly and use of an injectrode for precise drug delivery into targeted brain regions of an anesthetized rat, followed by monitoring of neuronal activity through recorded electrical signals.

쥐의 뇌에서 전기생리학적 기록을 통한 약물 미세주입

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review ...

JoVE Encyclopedia of Experiments

Neurophysiology

Electrophysiology Techniques

In Vivo Electrophysiological Recordings of Brain Activities from Multiple Sites in Rats

Source: Haumesser, J. K., et al. <a href="https://app.jove.com/v/55940">Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats.</a> J. Vis. Exp.(2017).
This video demonstrates the detailed procedure for preparing an anesthetized rat for electrophysiological recordings of brain activity from multiple sites. The scalp is dissected to access the skull, and holes are drilled at precise locations to install and secure electrodes. Tungsten microwire electrodes are then inserted into the targeted location to record the data.

쥐의 여러 부위에서 뇌 활동에 대한 생체 내 전기생리학적 기록

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

A Micro-drive Array Method for Electrophysiological Recording from Multiple Brain Regions

Source: Shikano, Y., et al. <a href="https://app.jove.com/v/56980">Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat.</a> J. Vis. Exp. (2018).
The video demonstrates simultaneous electrophysiological recording from multiple brain regions in a rat using an integrative micro-drive array. Recording electrodes are placed in the olfactory bulb, reference electrodes in the frontal cortex, and micro-drive array electrodes in the hippocampus. The rat is allowed to forage freely, and simultaneous recording from all the implanted electrodes is recorded using the array.