저자는, 드 라 공들인 실내 santé 퀘벡 (FRSQ), 건강 연구의 캐나다 연구소 (CIHR), 자연 과학 및 캐나다의 공학 연구위원회 (NSERC) Fonds에서 급여 상, 장비 보조금, 운영 자금을 인정하고 싶습니다 그리고 Concordia는 대학 연​​구 의자 프로그램 (CRUC)뿐만 아니라, 박사 제인 스튜어트에서이 원고에 대한 의견.

1. 동물 주택 <ol><li> 케이지 : 개별 쥐의 실행 륜 활동을 기록하기 위해, 각 케이지는 단일 쥐 및 실행 휠을 수용해야합니다. 실행 바퀴가 농축의 한 형태로 간주 될 수 있기 때문에, 모든 연구의 모든 동물은 실행 휠과 유사한 액세스 할 수 있어야합니다. </li><li> 침대 변경 : 그들은 동물과의 접촉을 최소화하기 때문에 동물 처리뿐만 아니라 새장 또는 침구의 변화는 모든 circadian 리듬 21-23에 비 빛 영향을 미칠 수 있으므로, 메쉬 - 바닥이있는 케이지가 이상적입니다. 이러한 트레이 시스템의 가용성에도 불구하고, 침구 변경 사항이 실험의 중요한 단계 중에 피해야한다. 대안이 더 드문 케이지 변경 사항을 허용 오래 지속 침구, 또는 의사 난수 일정에 변경 새장을 사용하여 포함되어 있습니다. </li><li> 절연 상자 : 연습장은 소리 감쇠, 빛 공동 아르 격리 상자에 보관해야합니다ntrolled, 그리고 환기가 잘되는. 격리 상자의 크기와 구성에 따라 각 상자 케이지의 수는 일반적으로 1-8까지 다양합니다. 단일 절연 상자에서 여러 새장을 주택 때, 하나는 다른 동물에서 나오는 각종 냄새와 소리가 개별 동물의 circadian 행동에 혼란 영향을 미칠 수 있다는 것을 알고 있어야한다. 이러한 문제를 방지하기 위해, 하나는 절연 상자 당 하나의 새장을 수용하려고 시도합니다. </li><li> 환기 : 적절한 공기 흐름이 상자 설치류 동물을위한 편안한 가정 환경 만들기에 필수적입니다. 각 상자의 팬 내부에 도달 상자 외부에서 빛을 방지 할 수 있도록, 후드해야합니다. 또한 팬들은 일반적으로 상자에서 공기를 제거하고 방에 날려 것입니다. 작은 빛 빡빡 통풍구는 공기가 여러 지점에서 격리 상자를 입력 할 수 있으며 불편 바람을 방지하는 데 도움이됩니다. 적절한 환기, 절연 상자 안에 온도가 (가 있다는 것을 확인하기 위해)에 걸리면 몇 시간 동안 폐쇄는가 설치된 방에있는 온도에 거의 동일해야합니다. </li><li> 조명 : 환경 광도 모든 케이지에서 동일해야합니다. 각 케이지 위의 비슷한 위치에서 하나의 빛을 정렬하고, 항상 같은 브랜드 / 전구 유형을 사용합니다. 케이지 수준에서 중간 강도 조명 (100-300 룩스)를 사용합니다. 오히려 circadian 시스템 이라기보다는 (예 : 마스킹)보다 빛에 기인 행동에 직접 변경 사항을 생산할 가능성이 더 높습니다 과도하게 높은 조명 수준을, 피하십시오. </li><li> 암흑 / 흐리게 붉은 색 조명 :이 어둠 속에서 동물 (상수 어둠 또는 밤에 등) 처리하거나 처리 할 필요가있는 경우, 나이트 비전 고글을 사용합니다. circadian 시스템이 붉은 파장 상대적으로 문자를 구분하기 때문에 또는, 희미한 붉은 조명을 사용할 수 있습니다. 사용 특정 붉은 빛이 그 alte하지 않는지 확인하기위한 테스트해야R 실행 - 휠 활동 (예 : 마스킹) 또는 circadian 시계를 조정 (예 : 위상 편이를 생성). </li></ol> 2. 데이터 수집 (그림 1 참조 - Vitalview 하드웨어 구성) <ol><li> 바퀴를 실행하면 : 직경 실행중인 휠의 인체 공학적는 사용 24의 양이 변경됩니다. 따라서, 마우스의 작고 가벼운 바퀴를 사용하며, 쥐에 대한 큰 무거운 바퀴. 바퀴를 세척하고 다시 설치하는 경우, 바퀴가 탁 트인, &quot;동요&quot;하지 않으며, 녹화 마이크로 스위치 휠의 각 회전에 의해 활성화되는 돌리고 할 수 있습니다 있는지 확인하십시오. </li><li> 마이크로 스위치 : 실행중인 휠의 각 혁명은 자기 또는 기계적 마이크로 스위치를 활성화해야합니다. 마이크로 스위치의 정보는 하나의 채널을 통해 전송하고 컴퓨터에 의해 기록 된 수 빈 시간이 지남에 따라 데이터 (예 : 매 2, 5, 6, 또는 10 분). </li><li> 컴퓨터 하드웨어 : 우리 runniNG 륜 녹음 Vitalview, 미니 Mitter (에 의해 개발 된 하드웨어 및 소프트웨어 플랫폼을 만들어 <a href="http://www.minimitter.com/vitalview_software.cfm" target="_blank">http://www.minimitter.com/vitalview_software.cfm</a> ). 그러나, 이러한 Actimetrics (에 의해 개발 ClockLab 같은 다른 녹음 플랫폼 있습니다 <a href="http://www.actimetrics.com/ClockLab/" target="_blank">http://www.actimetrics.com/ClockLab/가</a> ). 두 플랫폼은 하나의 컴퓨터 파일에 많은 단일 채널 소스에서 데이터를 (예를 들어 하나의 실행 휠에 의해 활성화 한 마이크로 스위치) 함께 가져. 개인 채널의 데이터는 그래프로 작성할 수 있으며 나중에 개별적으로 분석 할 수 있습니다. </li></ol> 3. 데이터 레코딩 <ol><li> 파일 : 위에서 언급 한 소프트웨어 플랫폼은 개별 파일은 각각의 실행 륜 기록이 생성되는 수 있도록 하나의 채널을 분리하는 데 사용할 수 있습니다. 이러한 데이터는 가장 시각화과 특별히 고안와 그래프이러한 Actiview (Minimitter, 벤드, OR), Circadia, 또는 모든 periodograms과 actograms를 생성 할 수 있습니다 Clocklab (Actimetrics, Wilmette, IL)와 같은 D 프로그램. 그러나, 단일 채널 파일도 열 및 엑셀 (마이크로 소프트, 레드몬드, WA)와 같은 일반적인 스프레드 시트 프로그램을 사용 분석 할 수 있습니다. </li><li> Circadian 시간 (중부 표준시) 계산 : 중부 표준시 12은, 정의에 의해, 야행성 동물에서 실행 륜 활동의 발병입니다. 24 시간 하루와 병렬로, 국제 대회에서 한 circadian 일 24 circadian 시간으로 나뉩니다. circadian 자유 실행 기간이 벽 시계에 의해 측정으로 24 시간 30 분 인 경우 따라서,, CT 0 CT12 후 약 12​​ 시간 15 분 발생합니다. </li></ol>

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관심 없음 충돌이 선언 없습니다.

실행 바퀴를 사용하여 매일 활동 리듬을 모니터링하는 것은 야행성 동물의 마스터 circadian 시계의 출력을 평가하기위한 가장 일반적으로 사용하고 신뢰할 수있는 방법입니다. 휠 - 실행 활동은, 그러나, 행동과 지속적으로 모니터링 할 수 있습니다 생리학의 여러 측면 중 하나입니다. 실행 륜 활동의 대부분은 밤에 발생하지만, 총 잠이 오지 않음의 30 % 이상이 낮 25,26 동안 발생합니다. 다?...

<table border="1"&gt;<tbody&gt;<tr&gt;<td<strong시약의&gt; 이름</strong</td&gt;<td<strong&gt; 회사 소개</strong</td&gt;<td<strong&gt; 카탈로그 번호</strong</td&gt;<td<strong&gt; 코멘트 (선택 사항)</strong</td&gt;</tr&gt;<tr&gt;<td&gt; Vitalview 카드 및 소프트웨어</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 855-0030-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; DP24 데이터 포트</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 840-0024-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; QA4 - 모듈</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 130-0050-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; 자기 스위치</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 130-0015-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; C-50 케이블 어셈블리</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 060-0045-10</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td휠 어셈블리를 실행&gt; 쥐</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 640-0700-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; 케이지와 트레이 지원</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 640-0400-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; 유용한은 새장을 떼어</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 664-2154-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td케이지에 대한&gt; 그리드 층</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 676-2154-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; 쓰레기 트레이</td&gt;<td&gt; 미니 Mitter</td&gt;<td&gt; # 684-2154-00</td&gt;<td&gt; (벤드, OR, USA)</td&gt;</tr&gt;<tr&gt;<td&gt; 램프 하우징</td&gt;<td&gt; Microlites 과학</td&gt;<td&gt; # R-101</td&gt;<td&gt; (토론토, ON, 캐나다)</td&gt;</tr&gt;<tr&gt;<td&gt; 4W 형광등 램프</td&gt;<td&gt; Microlites 과학</td&gt;<td&gt; # F4T5/CW</td&gt;<td&gt; (토론토, ON, 캐나다)</td&gt;</tr&gt;<tr&gt;<td&gt; 절연 방</td&gt;<td건축&gt; 사용자</td&gt;<td</td&gt;<td&gt; 28 &quot;높이 x 20&quot;W X 28 &quot;D ½&quot;검은 멜라민.</td&gt;</tr&gt;</tbody&gt;</table

recording and analysis of circadian rhythms in running-wheel activity in rodents

설치류가 자신의 홈 케이지에서 실행 휠을 무료로 이용하실 수 있습니다 때이 바퀴의 자발적인 사용은 일 1-5의 시간에 따라 달라집니다. 쥐, 햄스터, 그리고 마우스 등의 야행성 동물은 밤에 활성화되어 있으며 하루 동안 상대적으로 비활성. 많은 다른 행동과 생리 조치는 일상 리듬을 전시하지만, 동물에 실행 륜 활동이 마스터 circadian 시계, 시상 하부의 suprachiasmatic 핵 (SCN)의 출력 특히 안정적이고 편리한 측정 역할을합니다. 일반적으로 유입이라는 프로세스를 통해 실행 륜 활동의 일상 패턴은 자연 환경 빛 어두운주기 (; : 12 시간 - 어두운 예를 들어 12 시간 - 빛 LD 사이클)로 정렬됩니다. 그러나 circadian 리듬이 endogenously 일정한 어둠 속에서 ~ 24 시간 기간을 전시하고, 지속 행동의 패턴을 생성됩니다. 따라서, LD주기의 부재에, 실행 륜 활동의 기록 및 분석 할 수 있습니다시간의 일 주관적인를 결정하는 데 사용됩니다. 이러한 리듬은 circadian 시계가 감독하기 때문에 시간의 하루 주관적인는 circadian 시간 (중부 표준시)라고도합니다. 반면, LD주기가 존재할 때, 환경 LD주기에 의해 결정되는 시간의 하루는 zeitgeber 시간 (ZT)라고합니다. 실행 륜 활동에 circadian 리듬은 일반적으로 SCN 시계 6-8에 연결되어 있지만, 뇌와 몸 9-14의 많은 다른 지역에 circadian 발진기는 매일 활동 리듬의 규정에 관여 할 수있다. 예를 들어, 식품 예상 활동에 매일 리듬은 SCN 15,16을 필요로하지 않고 대신 추가 - SCN 발진기 17-20의 활동의 변화와 상관 있습니다. 따라서, 실행 륜 활동 녹음 마스터 SCN 시계의 출력에 대한뿐만 아니라, 매우 SCN 발진기의 활동에뿐만 아니라 중요한 행동 정보를 제공 할 수 있습니다. 우리는 descr 아래장비와 실험실 쥐에 circadian 전위의 활동 리듬을, 기록 분석하고 표시하는 데 사용할 방법을 ibe.

<ol><li> 컴퓨터 프로그램 : 전문 컴퓨터 프로그램은 일반적으로 actograms와 circadian 기간의 계산의 생성에 사용됩니다. 이 프로그램은 포함하지만, (Minimitter, 벤드, OR)와 Circadia Actiview에 제한되지 않습니다. </li><li> Actograms : Actograms가 실행 륜 활동의 일상 패턴 그래픽 그림을 제공합니다. 단일 해본 (x-축 = 24 시간) 두 번 해본 (x-축 = 48 시간) actograms가 있습니다. 두 가지 방법은 위에서 아래로 플롯...

Watch this Scientific Journal Video about Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents at JoVE.com

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

자원의 Circadian 리듬은 휠을 실행 포유류 활동은 단단히 뇌의 마스터 시계의 분자 진동에 커플 링된다. 따라서 행동이 일상 리듬이 circadian 시계의 기능에 대한 유전자, 약리, 환경 요인의 영향을 연구하는 데 사용할 수 있습니다.

쥐에 활동 륜 실행에 녹음 및 Circadian 리듬 분석

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

recording-analysis-circadian-rhythms-running-wheel-activity

Research

JoVE Journal

Neuroscience

21.1K 조회수.  McGill University. 자원의 Circadian 리듬은 휠을 실행 포유류 활동은 단단히 뇌의 마스터 시계의 분자 진동에 커플 링된다. 따라서 행동이 일상 리듬이 circadian 시계의 기능에 대한 유전자, 약리, 환경 요인의 영향을 연구하는 데 사용할 수 있습니다.

비디오: Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

Circadian 리듬을 연구 및 매개 변수 잠을 전위의 활동 시금 Drosophila</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

연구

신경과학

Measuring Circadian and Acute Light Responses in Mice using Wheel Running Activity

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2. They are responsible for timing our sleep/wake cycles and hormone secretion. Since this timing is not precisely 24-hours, it is synchronized to the solar day by light input. This is accomplished via photic input from the retina to the suprachiasmatic nucleus (SCN) which serves as the master pacemaker synchronizing peripheral clocks in other regions of the brain and peripheral tissues to the environmental light dark cycle3-7. The alignment of rhythms to this environmental light dark cycle organizes particular physiological events to the correct temporal niche, which is crucial for survival8. For example, mice sleep during the day and are active at night. This ability to consolidate activity to either the light or dark portion of the day is referred to as circadian photoentrainment and requires light input to the circadian clock9. Activity of mice at night is robust particularly in the presence of a running wheel. Measuring this behavior is a minimally invasive method that can be used to evaluate the functionality of the circadian system as well as light input to this system. Methods that will covered here are used to examine the circadian clock, light input to this system, as well as the direct influence of light on wheel running behavior.

This article will review methods that can be used to determine circadian function and light responsiveness in mice.

휠 실행 활동을 사용하여 마우스의 Circadian 및 급성 라이트 응답 측정

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2.  They ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

설치류를 행동에 이동 가능한 실리콘 프로브에 의한 뉴런의 대규모 녹화

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

여러인지 기능을 테스트 및 설치류에의 연결 활동을 녹음하기위한 완전 자동화된 고도 다목적 시스템

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

역행 형광 라벨이 확인 된 신경 세포의 표적 세포 외 단일 단위 기록을 허용<em&gt; 생체 내</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

메뚜기에 신경 활동을 특성화하기 위해 다중 단위 기록 방법 (<em&gt; Schistocerca 아메리카나</em&gt;) 강한 회로

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

설치류에 동시 fMRI를 깊은 뇌 자극

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the different organ systems from the periphery towards the central nervous system. As such, the increased activity or &#39;sensitization&#39; of mesenteric afferent nerves has been allocated an important role in the pathophysiology of visceral hypersensitivity and abdominal pain syndromes.
Mesenteric afferent nerve activity can be measured in vitro in an isolated intestinal segment that is mounted in a purpose-built organ bath and from which the splanchnic nerve is isolated, allowing researchers to directly assess nerve activity adjacent to the gastrointestinal segment. Activity can be recorded at baseline in standardized conditions, during distension of the segment or following the addition of pharmacological compounds delivered intraluminally or serosally. This technique allows the researcher to easily study the effect of drugs targeting the peripheral nervous system in control specimens; besides, it provides crucial information on how neuronal activity is altered during disease. It should be noted however that measuring afferent neuronal firing activity only constitutes one relay station in the complex neuronal signaling cascade, and researchers should bear in mind not to overlook neuronal activity at other levels (e.g., dorsal root ganglia, spinal cord or central nervous system) in order to fully elucidate the complex neuronal physiology in health and disease.
Commonly used applications include the study of neuronal activity in response to the administration of lipopolysaccharide, and the study of afferent nerve activity in animal models of irritable bowel syndrome. In a more translational approach, the isolated mouse intestinal segment can be exposed to colonic supernatants from IBS patients. Furthermore, a modification of this technique has been recently shown to be applicable in human colonic specimens.

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity.

마우스 Jejunal과 대장 세그먼트에서 장간막 구 심성 신경 활동의 생체 외 녹음에서

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic tectum, is a popular model to study how neural circuits self-assemble. The ability to carry out whole cell patch clamp recordings from tectal neurons and to record RGC-evoked responses, either in vivo or using a whole brain preparation, has generated a large body of high-resolution data about the mechanisms underlying normal, and abnormal, circuit formation and function. Here we describe how to perform the in vivo preparation, the original whole brain preparation, and a more recently developed horizontal brain slice preparation for obtaining whole cell patch clamp recordings from tectal neurons. Each preparation has unique experimental advantages. The in vivo preparation enables the recording of the direct response of tectal neurons to visual stimuli projected onto the eye. The whole brain preparation allows for the RGC axons to be activated in a highly controlled manner, and the horizontal brain slice preparation allows recording from across all layers of the tectum.

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

In this paper, we discuss three brain preparations used for whole cell patch clamp recording to study the retinotectal circuit of Xenopus laevis tadpoles. Each preparation, with its own specific advantages, contributes to the experimental tractability of the Xenopus tadpole as a model to study neural circuit function.

준비 및 Xenopus laevis Tectal 신경의 전체 셀 패치 클램프 기록에 대 한 프로토콜

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.