Name: in vitro recording of mesenteric afferent nerve activity in mouse jejunal and colonic segments
Uploaded: 2016-10-25T13:00:00
Description: Watch this Scientific Journal Video about In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments at JoVE.com

SN performed the experiments described above, performed the data analysis and drafted the manuscript. AD and JDM implemented the technique at our research facilities and aided in the data analysis. HC aided in performing the experiments. WJ, CK and DG assisted in implementing the afferent measurement technique in our lab, the data analysis and interpretation of the results. SF, JDM and BDW designed the study. All authors critically read and approved the final manuscript. SN is an aspirant of the Fund for Scientific Research (FWO), Flanders (11G7415N). This work was supported financially by the FWO (G028615N and G034113N).

아래에 설명 된 모든 동물 실험은 의료 윤리위원회와 앤트워프 대학 (파일 번호 2012-42)에 실험 동물의 사용에 의해 승인되었다. 1. Jejunal의 조직 준비 및 대장 구 심성 신경 <ol><li> jejunal 구 심성 신경의 준비 <ol><li> 지역 윤리위원회의 사전 실험에 승인 된 청소년 또는 성인 설치류의 쥐 안락사를 수행합니다 (예., 심장 천공, 자궁 경부 전위 등 뒤에 단말기 진정 ?...

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이 논문의 프로토콜은 우리 그룹과 다른 3,4,7,8,12,20,21,31에 의해 사용되는 설치류에 장간막 구 심성 신경의 활동을 연구하기 위해 재현 실험 방법을 설명합니다. 프로토콜 내에서 중요한 단계는 조직의 신속한 분리, 흡착 전극에 신경 가닥의 흡입 모세관으로 주변 지방 조직을 흡인하여 기관 조에서 유리 모세관의 적절한 &#39;밀봉&#39;를 포함한다. 유리 모세관의 개구가 정확하게 결정되어?...

감각 신호와 고통의 인식은 구 심성 신경, 척수 신경, 상승 및 하강 facilitatory 및 억제 경로 및 여러 가지 뇌 영역 사이의 복잡한 상호 작용의 결과 복잡한 과정이다. 따라서,이 수준 중 하나 이상에 변경 사항이 변경된 감각 신호와 질병 상태에서 내장 통증의 원인이 될 수 있습니다. 감각 신호 여러 기술이 모든 다양한 측면을 연구하기 위해 단일 세포 실험에 이르기까지 개발되고있다 (예를 들어, 신경 세포에 칼슘 이미징) 전체 동물 모델 (예를 들어, 같은 내장 운동 응답으로 행동 반응). 이 문서에 설명 된 기술은 연구자가 특별히 설치류에서 소장 또는 대장의 고립 된 세그먼트에서 시험관 내에서 구 심성 신경의 활동을 평가할 수 있습니다. 즉, 고립 위장 세그먼트는 (일반적으로 소장 또는 대장) 생리적 K와 관류 목적 내장 기록 챔버에 장착남군 솔루션입니다. 내장 신경 무연 해부 및 골반 내장의 또는 구심 신경의 구 심성 신경 활성의 등록을 허용하는 전극에 접속된다. 신경 활동 basally하거나, 또는 (점막)이 관내 관류 통해 기록 실 (serosally)에 직접 적용될 수 관내 압력 및 / 또는 약학 적 화합물의 증가에 응답 심성 방전 1-6에 미치는 영향을 평가하기 위해 기록 할 수있다 . 참고로, 내장의 신경은 원심성 섬유와 감각 구 심성에 추가 viscerofugal 구 심성이 포함되어 있습니다. 생체 내장 신경 기록의 주요 이점 중 하나는 연구자 신경 활성에 국소 도포 화합물의 직접적인 영향을 연구하기 위해 하나를 허용하는 중추 신경계에서 변조 또는 입력없이 신경 활성을 정량화 할 수 있다는 사실이다. 생체 내 방법을 사용하여 필요한만큼 또한, 중요 파라미터들의 모니터링은 N이다 (아래 참조)O 이상 관련. 시험관 내장의 기록이 최종적으로 더 적은 시간이 소요의 생체 대응보다. 이러한 점막 쓰다듬어 다른 자극, 본 프레이 모발을 사용하여 프로빙 또는 세그먼트의 연신에 응답하여 구 심성 신경의 활동은 창자 조직 아래 피닝과 대조적이다 (세로 방향으로 개방되는 개질 실험 구성에서 연구 될 수있다 로 이전 문제 7, 8에 설명 된)을 그대로 세그먼트를 사용하여 우리의 설치. 또한, 최근, 기술이 다시 길이 방향 세그먼트 (9)를 열어 아래로 고정하여 칼슘의 이미징 (imaging)을 통해 결장 벽 자체 결장 심성 신경 활성을 연구하기 위해 기술되었다. 이 생체 기술의 또 다른 버전은 척수에 구 심성의 항목 근처 신경 세포의 활성화를 측정 밖으로 구성되어 있습니다. 즉, 진정 동물이 발생하기 쉬운 위치, 전자에 위치건설 후궁 절제술의 수단으로 관심 프로젝트되는 구 심성 신경에 요천 추부 척수를 xposing 파라핀 충전 절개의 피부를 사용 백금 양극 전극 (10, 11)를 통해 지느러미 어린 뿌리를 드레이프 잘. 이 기술은 또한 연구자들은 얇게 유수 Aδ - 섬유 수초 C-섬유를 자신의 전도 속도에 따라 섬유의 특징과 구별 할 수 있습니다. 또한, 지느러미 작은 뿌리는 독점적으로 앞서 언급 한 혼합 구 심성 및 원심성 내장의 신경 대조적으로, 감각 구 심성 섬유를 포함하고있다. 절연 장 세그먼트에서 시험관 내에서 구 심성 신경 방전 기록은 독립적으로 인간의 절제에서 대장 구 심성 신경의 활동을 기록하는 최초의 남자 원고는 12, 13을 표본 게시 된 두 개의 연구 그룹으로, 인간의 표본을 사용하여 수행 할 수 있습니다. 이 기술의 구현을보다 용이하게 발생할 수 translati인간적 상태로 쥐의 데이터에, 연구자 쉽게 감응 감각 신경을 대상으로 약물을 식별 할 수 있습니다. 구 심성 신경의 활동뿐만 아니라 과대 구 심성 신경의 활성을 표적으로 새로운 치료 시약의 발견을 특성화의 임상 중요성은 정교 필드 14-19 많은 전문가들이 논의되어왔다. 상기 시험 관내 기술은 더 일반적 심성 신경 활성의 생체 측정에 공지 보완한다. 생체 내에서 신경 세포 활성도를 측정하는 동안, 신경 활성을 주목 세그먼트 삽관이어서 식별되는 동안의 마취 동물에서 직접 측정 할 수 있고, 유동 파라핀 채워진 웰 설치류 20 복벽 및 피부를 사용하여 구성된다. 관심의 구 심성 신경이어서 신경 활동 measuremen 있도록, 식별 단면 바이폴라 백금 전극 상에 배치되고티. 이 기술은 진정 동물이기는하지만 생활에 구 심성 신경 활성을 조절하는 연구를 할 수 있습니다; 예로서, 하나는 내강 팽창 또는 화합물의 정맥 내 투여와 같은 간섭에 응답 신경 활동을 연구 할 수있다. 중개 연구는 오늘날 주로 인간 유래 상청액의 적용에 중점 (예., 결장 생검 재배 말초 혈액 단핵 세포 등으로부터) jejunal 및 / 또는 구 심성 결장 마우스 (21, 22)에. 점막 응용 프로그램 대 장막의 차동 효과는 구 심성 신경 방전에 공부를 할 수 있도록 연구자가 직접 장기 욕조에 또는 장 세그먼트를 관류되는 관내 솔루션으로 하나 상층 액을 적용 할 수 있습니다. 이와 같이, 마우스 대장 구 심성, 기니 돼지 점막 밑 신경 세포 및 마우스 등의 루트에 과민 반응을 일으킬 수 과민성 대장 증후군 환자에서 그 대장 점막 조직 검사 supernatans를 줬습니다신경절 신경 세포 21,23,24. 마지막으로, 신경 세포의 활동을 기록하는 장간막 제한 및 / 또는 골반 신경은 위장관에 분포되어 있지 않습니다. 기타 신경 녹음 무릎 관절 (25)에 공급 구 심성에서 수행 될 수있는 다른 사람도 26-28 블래 구 심성 신경의 활동을 특징 짓는 반면, 증명 및 방광, 골반 심성뿐만 아니라 위장관 가능성 신경 결과 수렴한다는 증명 크로스 토크 29.

<table border="0" cellpadding="0" cellspacing="0" width="788">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">sodium chloride (NaCl)</td>
			<td style="width:247px;">VWR Chemicals</td>
			<td align="right" style="width:119px;">27,810,295</td>
			<td style="width:167px;">compound Krebs solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">potassium chloride (KCl)</td>
			<td style="width:247px;">Acros organics</td>
			<td align="right" style="width:119px;">196770010</td>
			<td>compound Krebs solution</td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:255px;">sodium dihydrogen phosphate (NaH2PO4)</td>
			<td style="width:247px;">VWR Chemicals</td>
			<td align="right" style="width:119px;">1,063,461,000</td>
			<td>compound Krebs solution</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:255px;">sodium bicarbonate (NaHCO3)</td>
			<td style="width:247px;">Merck</td>
			<td align="right" style="width:119px;">1,063,291,000</td>
			<td>compound Krebs solution</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:255px;">magnesium sulfate (MgSO4)</td>
			<td style="width:247px;">Merck</td>
			<td align="right" style="width:119px;">1,058,861,000</td>
			<td>compound Krebs solution</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:255px;">calcium chloride (CaCl2)</td>
			<td style="width:247px;">Merck</td>
			<td align="right" style="width:119px;">23,811,000</td>
			<td>compound Krebs solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">D-glucose</td>
			<td style="width:247px;">VWR Chemicals</td>
			<td style="width:119px;">1011175P</td>
			<td>compound Krebs solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Distilled water</td>
			<td style="width:247px;"></td>
			<td style="width:119px;"></td>
			<td>compound Krebs solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">PVC tubing</td>
			<td style="width:247px;">Scientific Laboratory Supplies</td>
			<td style="width:119px;"></td>
			<td>The intestinal segment should be mounted over PVC tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Silicone tubing</td>
			<td style="width:247px;">Scientific Laboratory Supplies</td>
			<td style="width:119px;"></td>
			<td>The rest of the tubing, ideally silicone-based - more easily dislodging of debris in the tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Silk thread</td>
			<td style="width:247px;">Pearsall Limited</td>
			<td style="width:119px;">10B15S220</td>
			<td>Attachment of the segment over the PVC tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Syringe driver</td>
			<td style="width:247px;">Harvard Apparatus</td>
			<td style="width:119px;">55-2222</td>
			<td>Intraluminal infusion of Krebs</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:255px;">Binocular - including 10x magnification in oculair</td>
			<td style="width:247px;">Zeiss STEMI 2000</td>
			<td style="width:119px;"></td>
			<td>Optimal visualization for the dissection of the afferent nerve</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Homeothermic Blanket Control Unit</td>
			<td style="width:247px;">Harvard Apparatus</td>
			<td align="right" style="width:119px;">507214</td>
			<td>Heating of the organ chamber</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:255px;">Custom made organ bath with Sylgard covered bottom</td>
			<td style="width:247px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Spike2 software</td>
			<td style="width:247px;"></td>
			<td style="width:119px;"></td>
			<td>Recording and analysis of the data</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:255px;">Insect pins, 500 pieces, stainless steel, diameter 0.2 mm</td>
			<td style="width:247px;">Austerlitz insect pins minutiens</td>
			<td style="width:119px;"></td>
			<td>Dissection of the afferent nerve</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Tweezer Dumont #5 inox 11cm</td>
			<td style="width:247px;">World Precision Instrument</td>
			<td align="right" style="width:119px;">500341</td>
			<td>Dissection of the afferent nerve</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Scissors, spring, 14 cm</td>
			<td style="width:247px;">World Precision Instrument</td>
			<td align="right" style="width:119px;">15905</td>
			<td>Dissection of the afferent nerve</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">DB digitimer&#160;</td>
			<td style="width:247px;">NL 108T2/10</td>
			<td style="width:119px;"></td>
			<td>pressure transducer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Micromanipulator</td>
			<td style="width:247px;">Narishige</td>
			<td style="width:119px;">M-3333</td>
			<td>3D manipulation of the suction electrode</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Micromanipulator</td>
			<td style="width:247px;">X-4 rotating block</td>
			<td style="width:119px;"></td>
			<td>3D manipulation of the suction electrode</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Micromanipulator</td>
			<td style="width:247px;">GJ-8 magnetic stand</td>
			<td style="width:119px;"></td>
			<td>3D manipulation of the suction electrode</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">LightSource</td>
			<td style="width:247px;">Euromex Microscopes Holland EK-1</td>
			<td style="width:119px;"></td>
			<td>Optimal visualization for the dissection of the afferent nerve</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">CED 1401 Recording Apparatus</td>
			<td style="width:247px;"></td>
			<td style="width:119px;"></td>
			<td>Recording of afferent nerve activity</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Humbug 50/60Hz Noise Eliminator</td>
			<td style="width:247px;">Quest Scientific Instruments</td>
			<td style="width:119px;"></td>
			<td>Elimination of background noise</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Infusion Pump</td>
			<td style="width:247px;">Gibson Minipuls 2</td>
			<td style="width:119px;"></td>
			<td>Infusion of the organ chamber in which the segment is mounted</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:255px;">Microelectrode Holder Half Cells 1.5 mm</td>
			<td style="width:247px;">World Precision Instrument</td>
			<td style="width:119px;">MEH2SW</td>
			<td>Suction electrode for isolation of the afferent fiber</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:255px;">Borosilicate Glass Capillaries, 300 pc; 1.5/0.84 OD/ID</td>
			<td style="width:247px;">World Precision Instrument</td>
			<td style="width:119px;">1B150-4</td>
			<td>Capillary for the isolation of the afferent nerve</td>
		</tr>
	</tbody>
</table>

in vitro recording of mesenteric afferent nerve activity in mouse jejunal and colonic segments

구 심성 신경은 정상적인 생리학에 관한 정보를 전달할뿐만 아니라, 방해 항상성 및 중추 신경계 향해 주변으로부터 다양한 기관계의 병리 생리 학적 과정의 신호뿐만 아니라. 이와 같이, 증가 된 활성 또는 장간막 심성 신경 &#39;증감&#39;내장 과민 복통 증후군의 병태 생리에서 중요한 역할을 할당하고있다. 장간막 구 심성 신경의 활동 목적 내장 기관 욕에 장착되고 이로부터 내장 신경은 연구자 직접 위장 세그먼트에 인접한 신경 활성을 평가할 수 있도록 격리 된 격리 장 세그먼트 시험 관내에서 측정 할 수있다. 활성은 세그먼트의 팽창 또는 전달 serosally intraluminally 또는 약학 화합물의 첨가 동안 다음, 표준 조건의 기준을 기록 할 수있다. 이 기술은 수연구자 쉽게 제어 표본 말초 신경계를 대상으로 약물의 효과를 연구하는 단계; 게다가, 그것은 질병 중에 변경하는 방법 신경 활동에 대한 중요한 정보를 제공합니다. 이는 구 심성 신경 소성 활성을 측정하는 것은 전용 캐스케이드 시그널링 복잡한 신경 하나의 중계국을 구성하고, 연구자들은 다른 레벨 (예컨대, 후근 신경절, 척수 또는 중추 신경계에서 신경 세포 활성도를 간과 숙지하지 부담해야하지만 유의해야 ) 완전히 건강과 질병의 복잡한 신경 생리학을 명료하게하기 위해서입니다. 일반적으로 사용되는 응용 프로그램은 리포 폴리 사카 라이드의 관리에 응답하여 신경 활동의 연구 및 과민성 대장 증후군의 동물 모델에서 구 심성 신경 활동의 연구를 포함한다. 더 병진 방법에서, 고립 된 마우스 장 세그먼트는 IBS 환자로부터 대장 상층 액에 노출 될 수있다. 또한, 변형이 기술의 최근 인간 결장 시험편에 적용하는 것으로 나타났다.

Jejunal 심성 신경 활성을 기준으로 측정 한 응답 OF-1 생쥐 9 8 주 된 수컷의 팽창을 진입로. 물과 일반 우를 활용할 무제한 액세스 할 수있는 동물은 표준화 된 조건 그룹 (50 %, 12 시간 명암주기 - - 22 ° C, 습도 40 케이지 당 6 동물 20)을 수용했다. 마우스의 Jejunal 세그먼트 (자발적인 활동을 11.47 ± 3.31 꼬마 도깨비 / 초를 의미) 0 mmHg로의 관내 압력 기준에서 불규칙한 자연 구 심?...

Watch this Scientific Journal Video about In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments at JoVE.com

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 ...

The overall goal of this technique is to record in vitro splanchnic afferent nerve activity from a single jejunal or colonic nerve bundle in response to ramp distensions and the application of compounds. This method can help answer key questions in the field of neuroscience, such as whether afferent nerves supplying the gastrointestinal tract are hypersensitized or desensitized during various disease states. The main advantages of this technique are that it allows nerve discharge recording in vitro and that it allows to quantify the nerve activity without modulation or inputs of the central nervous system.The implication of this technique extends toward the therapy or diagnosis of several pain syndromes, such as the irritable bowel syndrome, since nerve sensitization plays a major role in the pathogenesis. Generally, individuals new to this method will struggle because the dissection of nerve bundles and the single unit analysis of the recorded signal will take a substantial amount of training. To begin this procedure, place the sacrificed mouse in the supine position.Perform an abdominal midline incision through the skin and abdominal muscle layer using a scalpel extending from the xiphoid process to the pubic bone. Afterward, rinse the abdominal cavity with cold Krebs solution in order to prevent the intraabdominal tissues from drying out. Then rapidly excise the entire jejunum using sharp scissors by removing approximately 20 centimeters of the small bowel, starting immediately from the duodenojejunal flexure.Be careful not to damage the surrounding structures, and keep the bowel's mesentery, which contains jejunal blood vessels and afferent nerves, intact. Next, place the excised jejunum in cold Krebs solution on ice while oxygenating it continuously with Carbigen. Then, cut the jejunum in approximately three centimeter long loops.Locate the mesenteric bundle containing the vessels and splanchnic nerve near the center of the respective loop in order to facilitate the mounting of the loop in the recording chamber. Subsequently flush each segment with Krebs solution using a blunt catheter to remove the luminal contents and chyme, as they contain digestive enzymes that will accelerate the deterioration of the tissue sample in vitro. After that, select a segment for measuring the afferent nerve activity and place it in the recording chamber coded with a silicone elastomer layer.Keep the Krebs temperature in the recording chamber at 34 degrees Celsius. Then mount the jejunal segment in the organ bath, so that the oral end is connected to the syringe driver providing luminal flow and the aboral end connects to the outflow. Slightly stretch the segment, but take care not to exert excessive tension.Attach both ends firmly using silk ligatures to the in and outflow ports. Afterward, attach the syringe driver to the oral end and perfuse the jejunal segment intraluminally with Krebs solution at 10 milliliters per hour. Pin the mesentery of the mounted intestinal segment flat against the silicone elastomer bottom layer.Then, stretch the mesentery out in order to optimize the visualization of the mesenteric bundle. Next, perform a test on ramp distension by closing the output port until the intraluminal pressure of the intestinal segment reaches 60 millimeters of mercury in order to verify that no intraluminal Krebs solution is leaking from the mounted segment. Observe a smooth rise in the intraluminal pressure without interruptions.Expect small contractions of the segment during the initial distension phase. If required, block the peristaltic activity by adding one micromole of the L-type calcium channel blocker nifedipine to the Krebs solution. Under a stereo microscope, gently start to peel off the fat tissue from the mesentery by gently tugging it with two small tweezers, taking care not to damage the vessels and the afferent nerve that are buried in the fat tissue.Start peeling off the fat tissue at a distance from the jejunum to expose both blood vessels in the mesenteric bundle. Identify the afferent jejunal nerve in between both vessels as a thin white thread encapsulated in the adipose tissue. Then dissect it more proximally toward the jejunum by gently peeling the fat tissue away using tweezers.Next, dissect the jejunal mesenteric nerve of the segment by removing the adipose tissue adherent to the nerve. Transect the nerve using sharp tissue scissors. If required, peel off the remaining fat and connective tissue as well as the epineural sheath by gently tugging it.In this procedure, using a micromanipulator lower the tip of the suction electrode into the organ bath. Then, gently aspirate some Krebs solution from the organ bath, so that the tip of the electrode is submerged in the Krebs solution. Ensure that the Krebs solution covers the wire electrode inside the suction electrode.Next, position the tip of the suction electrode immediately next to the transected afferent nerve strand and draw the transected nerve strand into the capillary over its entire length. Maneuver the tip of the electrode towards the adipose tissue, and aspirate it into the glass capillary with the plunger, thereby mechanically sealing the nerve in the capillary from the contents of the organ bath. The adequate sealing of the glass capillary from the recording chamber by aspirating some surrounding fat tissue into the capillary is pivotal in order to minimize the redundant background noise that will ultimately hamper the final analysis.To verify the recording of afferent nerve activity, perform a ramp distension induced increase in afferent firing by closing the output port to lead to a gradual rise in pressure in the intestinal segment. Only perform the desired experimental protocol when three consecutive ramp distensions yield a reproducible multi-unit discharge. Following nerve isolation, stabilize the preparation for 15 minutes in order to obtain steady spontaneous afferent nerve activity before performing the actual experiments.Shown here is a schematic representation of different afferent fiber units based on their mechanosensitive profile. Low threshold fibers predominantly display an increased nerve activity at low distension pressures, resulting in an LT percentage of over 55%High threshold units on the contrary only display an increase in firing rate at noxious pressures. Wide dynamic range fibers display a gradual increase in nerve activity during the entire distension, whereas mechanically insensitive fibers do not respond to increasing distension pressures.This figure shows the mesenteric afferent nerve discharge in the wild type mice during ramp distension. Note that in the biphasic distension profile the initial rise in nerve discharge is due to low threshold and wide dynamic range fibers, whereas high threshold and wide dynamic range fibers contribute to the second increase in discharge. Once mastered, the isolation of the jejunal nerve and initial baseline recording can be done in less than one hour if performed properly.More time is necessary when studying the effect of compounds administered into the organ bath. While attempting this procedure, it's important to remember to maintain a well standardized approach, such as consistently identifying the same segment for all experiments. Following this procedure, other methods like the isolation of dorsal root ganglia with subsequent calcium imaging can be performed in order to assess which relay stations in the gastrointestinal nervous system are involved in the pathogenesis of several diseases.After its developments, this technique paved the way for researchers in the field of neuroscience to explore the pathogenesis of several pain syndromes in gastrointestinal research. After watching this video, you should have a good understanding of how to isolate jejunal afferent nerves from the mouse gastrointestinal tract, and record activity from it during ramp distensions.

Research

JoVE Journal

Neuroscience

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

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

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

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 ...

연구

신경과학

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

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<em&gt; 체외</em근육의 구 심성의 소성 특성을 연구&gt; 성인 마우스 근육 - 신경 준비

Muscle sensory neurons innervating muscle spindles and Golgi tendon organs encode length and force changes essential to proprioception. Additional ...

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마우스 헤어 난포 피침 엔딩에서 기계적으로 자극 된 수입 성의 산출과 신경 터미널 라벨의 결합 기록

A novel dissection and recording technique is described for monitoring afferent firing evoked by mechanical displacement of hairs in the mouse pinna. The ...

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In Vitro Characterization of the Electrophysiological Properties of Colonic Afferent Fibers in Rats

생체 외에서 쥐에 있는 저 승 구심 성 섬유의 Electrophysiological 속성의 특성

Dysfunction of the colonic sensory nerves has been implicated in the pathophysiology of several common conditions, including functional and inflammatory ...

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

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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) ...

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체외에서 감각 및 모터 신경에서 섬유 아세포와 슈완 세포의 정화

The principal cells in the peripheral nervous system are the Schwann cells (SCs) and the fibroblasts. Both these cells distinctly express the sensory and ...

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Neurofilament protein polymers move along axons in the slow component of axonal transport at average speeds of ~0.35-3.5 mm/day. Until recently the study ...