본 연구는 박사 바이 - 추앙 Shyu (NSC 99-2320-B-001-016-MY3, NSC 100-2311-B-001-003-MY3 및 NSC 102-2320-에 국가 과학위원회의 보조금 지원 B-001-026-MY3). 이 작품은 중앙 연구원에서 자금을받은 생명 과학 연구소에서 실시 하였다.

본 연구의 프로토콜은 대만의 중앙 연구원 기관 동물 관리의 승인 및 이용위원회를 받았다. 1. 동물 준비 <ol><li> (- 400g 약 300) 수컷 흰쥐를 가져옵니다. 음식과 물을 무료로 이용할 수로 (오전 8:00에 불이) 12 시간 / 12 시간 빛 / 어둠 사이클에서 - (22 °의 C, 습도 50 % 21) 에어컨 방에 ​​쥐를 유지한다. </li></ol> 2. 실험 절차 <ol><li> 쥐의 모든 실험 전에 1 주간 자신의 홈 케이지의 환경에 적응하도록 허용합니다. 적응하는 동안, 기준을 설정하는 데 프레이와 발바닥 테스트를 수행합니다. </li><li> 프레이 테스트 <ol><li> 거주 30 분 동안 아크릴 케이스 (30 cm × 30 cm × 80 cm)에 쥐를 놓습니다. </li><li> (. 아니 필라멘트 11-20) 본 프레이 필라멘트를 구하는 동일한 길이를 가지고 있지만, 직경이 변화하는 범위를 제공하도록100g - 2의 힘. </li><li> 쥐의 뒷발의 발 움츠림 역치를 평가 5 분 intertrial 간격 아크릴 판에 그물 형 포트를 통해 hindpaws의 중심을 자극 프레이 필라멘트를 사용한다. 최대인가 된 압력이 기록 될 때까지 로우에서 하이로 오름차순으로 필라멘트를 사용하여, 최대인가 된 압력을 기록한다. (10) </li><li> 쥐 자극에 대한 발 움츠림 반응을 나타낼 때, 필라멘트 수를 기록한다. 가장 낮은 자극에 따라이 재판에서 철수 응답을 확인합니다. </li><li> 동일한 래트에 연속 3 회, 총 즉시 동일한 절차를 반복한다. (g)에 대응하는 힘에 필라멘트 번호를 변환하고 값을 평균. </li></ol></li><li> 발바닥 테스트 <ol><li> 거주 30 분 (4 개의 프레임 80 cm × 30 cm × 15 센티미터로 구분) 투명한 플렉시 글라스 상자에 쥐를 놓습니다. </li><li> INF를 사용하여rared 광선은 유리판을 통해 뒷발의 중심을 자극한다. 약 10 초의 평균 발 움츠림 응답 대기 시간을 얻기 위해 적외선 강도를 조정한다. 적외선 광원을 켜고 디지털 솔리드 스테이트 타이머를 시작하는 키를 누름으로써 시험을 실시한다. 적외선 광선의 지속 시간을 조작 할 수 있습니다. </li><li> 래트가 발 움츠림 반응을 나타낼 때에 ​​외광의 지속 시간을 기록한다. 최장 기간은 조직의 손상을 방지하기 위해 각 시험에서 20 초를 초과 할 수 없습니다. 연속적인 자극을 피하기 위해 적어도 5 분의 intertrial 간격을 사용합니다. </li><li> 좌우 hindpaws 세 실험 테스트를 반복하고, 각 래트 뒷발에 대한 각각의 평균을 계산한다. </li></ol></li><li> 콜라게나 병변 수술 <ol><li> 수술 자극이 발생에 발가락 - 핀치 응답의 손실과 체세포 응답 할 때까지 4 % 이소 플루 란과 쥐를 마취. 2 % isoflur - 1.5 마취 유지수술 시간 동안 메탄. </li><li> 37.5 °의 C - 36.5에서 체온을 유지하기 위해 간단한 가열 패드와 정위 장치에서 쥐를 놓습니다. 마취 동안 건조를 방지하기 위해 눈에 아이 크림을 적용합니다. </li><li> 멸균 전기 면도기로 털을 면도하고 두피의 정중선을 따라 메스와 부드러운 절개 (약 2-2.5 cm)를 확인합니다. 섭리의 요오드 및 소독 75 % 알코올 용액을 교대로 피부와 두개골을 청소합니다. 수술 단계 동안 모든 기기, 도구 살균 75 %의 알코올을 사용하고, 작업대는 무균 상태를 유지하기. 그 전에, 모든 수술 재료는 증기 오토 클레이브 멸균해야합니다. </li><li> (시상의 (복부 후 내측 시상 핵 [VPM]와 복부 외측 시상 핵 [VPL] 포함) VB를 통해 전기 드릴로 두개골에 3.0 작은 구멍 (3 mm 직경)를 멸균 장갑과 악기를 사용하여 드릴 -3.5 mm 후방, 3.0-3.5 mm 측면) 정수리, 두개골 표면 5.5-5.8 mm 깊이합니다. </li><li> Microinject 생리 식염수 0.5 μL 또는 각각 제어 및 실험 그룹에서 4 형 콜라겐 용액 0.125 U. </li><li> 약물의 확산을 허용하도록 추가로 5 분 동안 곳에 주사 바늘을 유지. </li><li> 두개골에 구멍을 채우기 위해 치과 시멘트를 사용하고 절개를 봉합. 봉합 절개 후 상처 로컬 진통 마취제 (리도카인 연고)를 적용하고, 자신의 홈 케이지로 돌아갑니다. </li><li> 수술 후 단독으로 그들은 흉골 드러 누움을 유지할 때까지 플라스틱 케이지의 쥐를 수용하고, 마취에서 회복 될 때까지 따뜻하게 유지합니다. </li></ol></li><li> 수술 회복 후 행동 테스트 <ol><li> 수술에서 회복 7 일 후, 섹션 테스트 단계 2.2과 2.3의 절차를 반복합니다. 건강과 동물의 발달 상태를 모니터링하면서 사주를 통해 행동 테스트를 수행합니다. </li><li&gt; 이후 수술 단계의 제어 및 실험군의 동물의 건강 상태 (예를 들어, 체중, 먹이 양, 그리고 자유로운 움직임)을 모니터한다. </li></ol></li><li> PE-50 튜브와 함께 정맥 주입을 수행 <ol><li> 4 % 이소 플루 란 마취 래트 및 36.5에서 체온을 유지 - 단순한 가열 패드와 37.5 ° C. </li><li> 메스로 각각 왼쪽 어깨와 흉강의 복부 부분의 지느러미 앞발의 일부 교차로의 중간 선에 두 개의 구멍 (2cm 직경 각)을 잘라. </li><li> 두 구멍 사이 가위로 피부와 근육을 떼어 놓다. </li><li> 복부 구멍을 통해 외부 경정맥에 PE-50 튜브 (20cm 길이)의 한쪽 끝을 연결합니다. 지느러미 구멍 같은 PE-50 튜브의 말단을 연결하고, 피부에 부착합니다. </li><li> 확인하기 위해 경정맥에 식염수를 주입하는 주사기를 사용하여 그 PE-50 튜브방해하지 않습니다. </li><li> 절개 부위를 봉합하고, 6 ㎎ / ㎏ 겐타 마이신 (복강 내)로 쥐를 주입. </li><li> 20 U / ㎖ 헤파린과 생리 식염수 0.1 ml의 다음에, 0.3 ml의 0.9 %의 식염수로 튜브를 수술 후 매일을 플래시합니다. </li></ol></li><li> 최종 행동 테스트 <ol><li> 반복이 그 동작을 확인 2.2-2.3 단계 단계 2.6 이후 1 주일 수술 복구 후 단계 2.5에 비해 안정적이다. </li></ol></li><li> 추적자 주입 <ol><li> 적응을위한 10 분 - 5 휴식 케이지에 쥐를 놓습니다. </li><li> 스플리터를 사용하여, 두 1ml를 주사기에 PE-50 튜브를 연결합니다. 생리 식염수 하나의 주사기를 입력하고 (- 0.5 ml의 0.3의 볼륨에 125 μCi를 / kg) [14 C] -IAP 솔루션을 다른 입력합니다. </li><li> 외부 경정맥에 추적자를 주입하고, 3 M 염화칼륨 가득 다른 주사기 주사기를 교체합니다. </li><li> 텐 초 추적자의 INJ 후ection, 표준 안락사 방법에 따라 동물을 이소 플루 란의 과다 복용에서 3 M 염화칼륨을 주입하고, 희생 (즉, 50 분 동안 기화기에서 이소 플루 란) 대만하여 중앙 연구원 기관 동물 관리의 지침과 이용위원회를 기반으로. </li><li> 1 분 후, 두개골을 노출하고, 나머지 근육 잘라내. rongeurs를 사용하여 뇌에서 두개골의 지느러미 표면을 벗겨. rongeurs을 이용하여 두개골의 측면을 얻어 낸다. 다음에, 주걱을 이용하여, 뇌의 복부 표면을 따라 후각 신경 전구 연결을 잘라 뇌를 제거한다. </li><li> 드라이 아이스와 메틸 부탄의 두뇌를 동결 최적의 절삭 온도 OCT () 화합물을 사용하여 (약 -55 ° C). 냉동실에 뇌 조직을 저장합니다. 11, 12 </li></ol></li><li> 뇌 슬라이스 <ol><li> 후뇌이 아래를 향하도록하여, 마이크로톰에서 조직의 방향. 20로 뇌를 슬라이스 그라 이오 스탯을 사용하여# 181; m 두께의 섹션. </li><li> 각 조각 사이에 240 μm의 간격으로, -20 ° C에서 저온 유지 장치에 현미경 슬라이드에 뇌 조각을 놓습니다. </li><li> 현미경 슬라이드와 -20 ° C에서 3 일간 노출 카세트에 등급 방사능 다섯 표준 필터 종이를 놓습니다. 뇌 조각의 순서에 따라 위에서 아래로 현미경 슬라이드를 준비. 마지막으로, 카세트의 저부에 여과지를 배치했다. 12 </li><li> 노광 카세트로부터 형광면을 제거하고 뇌 슬라이스 [14 C] -IAP 흡수를 보여주기 위해 이미지를 형광체 스크린을 읽고 생성하는 가변 모드 이미 저를 사용한다. </li></ol></li></ol> 3. 데이터 분석 <ol><li> 단계 2.9.4 후에 통계적 매개 변수 사상 (SPM) 및 ImageJ에 소프트웨어를 사용하여 이미지를 조정한다. 직렬 관상 섹션을 사용하여 모든 이미지를 재구성. 부드럽고 기준 쥐 뇌 모델에 따라 이미지를 정규화.12 13 </li><li> 정량적 평가를 위해, 픽셀 신호 강도를 판정하고, 통계 해석 소프트웨어를 사용 ImageJ에 소프트웨어를 사용하여 관심 영역 뇌 영상 (ROI)을 측정한다. (12, 13) </li><li> 다른 뇌 핵 사이의 연결을 조사하기 위해, 지역간 상관 행렬의 방사능 비율을 표시하는 MATLAB 상관 분석 소프트웨어를 사용하여 컬러 맵 행렬로 시각화. 마지막으로, 네트워크 분석 Pajek 소프트웨어를 사용한다. (12) (13) </li><li> 베이스 라인에서 가짜 및 CPSP 그룹의 프레이 시험에서 발바닥 시험 및 기계적 힘에 열 관용의 지속 기간을 비교, 2 × 편차의 5 양방향 혼합 분석 (ANOVA), 그룹 및 주 요인으로 사용하고 주 1 - 5. ANOVA 그룹 뇌 영역에있어서 방사능 비율을 측정하는 2 × 31 양방향. 적절한 경우, Tukey에의 정직 유의 한 차이 (HSD) 사후 검사를 실시하고 있습니다. 칼culate 피어슨 상관 관계는 가짜와 CPSP 그룹에서 선택된 뇌 영역의 모든 사이에 상관 관계를 평가하는 계수. </li></ol>

<ol>
	<li>Finnerup, N. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+central+neuropathic+pain+states.">A review of central neuropathic pain states.</a> Curr Opin Anaesthesiol. 21 (5), 586-589 (2008).</li><li>Andersen, G., Vestergaard, K., Ingeman-Nielsen, M., Jensen, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence+of+central+post-stroke+pain.">Incidence of central post-stroke pain.</a> Pain. 61 (2), 187-193 (1995).</li><li>Chen, B., Stitik, T. P., Foye, P. M., Nadler, S. F., DeLisa, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+post-stroke+pain+syndrome:+yet+another+use+for+gabapentin?.">Central post-stroke pain syndrome: yet another use for gabapentin?.</a> Am J Phys Med Rehabil. 81 (9), 718-720 (2002).</li><li>Greenspan, J. D., Ohara, S., Sarlani, E., Lenz, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allodynia+in+patients+with+post-stroke+central+pain+(CPSP)+studied+by+statistical+quantitative+sensory+testing+within+individuals.">Allodynia in patients with post-stroke central pain (CPSP) studied by statistical quantitative sensory testing within individuals.</a> Pain. 109 (3), 357-366 (2004).</li><li>Klit, H., Finnerup, N. B., Jensen, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+post-stroke+pain:+clinical+characteristics,+pathophysiology,+and+management.">Central post-stroke pain: clinical characteristics, pathophysiology, and management.</a> Lancet Neurol. 8 (9), 857-868 (2009).</li><li>Kumar, G., Soni, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+post-stroke+pain:+current+evidence.">Central post-stroke pain: current evidence.</a> J Neurol Sci. 284 (1-2), 10-17 (2009).</li><li>Denk, F., McMahon, S. B., Tracey, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pain+vulnerability:+a+neurobiological+perspective.">Pain vulnerability: a neurobiological perspective.</a> Nat Neurosci. 17 (2), 192-200 (2014).</li><li>Ji, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cognitive+impairment+in+pain+through+amygdala-driven+prefrontal+cortical+deactivation.">Cognitive impairment in pain through amygdala-driven prefrontal cortical deactivation.</a> J Neurosci. 30 (15), 5451-5464 (2010).</li><li>Jungehulsing, G. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Levetiracetam+in+patients+with+central+neuropathic+post-stroke+pain:+a+randomized,+double-blind,+placebo-controlled+trial.">Levetiracetam in patients with central neuropathic post-stroke pain: a randomized, double-blind, placebo-controlled trial.</a> Eur J Neurol. 20 (2), 331-337 (2013).</li><li>Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M., Yaksh, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+tactile+allodynia+in+the+rat+paw.">Quantitative assessment of tactile allodynia in the rat paw.</a> J Neurosci Methods. 53, 55-63 (1994).</li><li>Jay, T. M., Luciqnani, G., Crane, A. M., Jehle, J., Sokoloff, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+local+cerebral+blood+flow+with+[14C]iodoantipyrine+in+the+mouse.">Measurement of local cerebral blood flow with [14C]iodoantipyrine in the mouse.</a> J Cereb Blood Flow Metab. 8 (1), 121-129 (1988).</li><li>Lu, H. C., Chang, W. J., Kuan, Y. H., Huang, A. C., Shyu, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+[14C]iodoantipyrine+study+of+inter-regional+correlations+of+neural+substrates+following+central+post-stroke+pain+in+rats.">A [14C]iodoantipyrine study of inter-regional correlations of neural substrates following central post-stroke pain in rats.</a> Mol Pain. 11 (1), (2015).</li><li>Wang, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+brain+activation+during+retrieval+of+visceral+pain-conditioned+passive+avoidance+in+the+rat.">Functional brain activation during retrieval of visceral pain-conditioned passive avoidance in the rat.</a> Pain. 152, 2746-2756 (2011).</li><li>Wasserman, J. K., Koeberle, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+a+hemorrhagic+rat+model+of+central+post-stroke+pain.">Development and characterization of a hemorrhagic rat model of central post-stroke pain.</a> Neuroscience. 161 (1), 173-183 (2009).</li></ol>

저자는 공개 아무것도 없어.

행동 검사에서 CPSP 그룹 기준의 본 프레이 시험 열 통증 검사 기계적 힘의 발 움츠림 역치의 감소 및 일주 전시 -. 5. 결과는 이전의 연구와 일치 하였다 14 에서 [14 C] -IAP있어서 다른 뇌 조각의 정량적 분석을 위해 뇌 영상의 화소 강도에 의존한다. 뇌 영상의 데이터를 평가하기 위해, 픽셀 신호 강도를 정의 하였다. 46.​​979 CPM - 본 연구에서, 환경 배경 신호는...

스트로크 출혈이 신경 병증 성 통증으로 고통 환자의 8 % 이상에서 발생하는 것으로 나타났다과 같은 중앙 뇌졸중 후 통증 (CPSP)라고합니다. 1-3 CPSP가 체 감각 장애, 이에 과민과 이질통을 유도 발생할 수 있습니다. (4) 그러나 , CPSP의 체성 감각 기능 장애의 병태 생리 학적 메커니즘은 불확실 남아있다. 예를 들어, 체성 감각의 손실은 출혈성 뇌 영역에서 신경 deafferentation 기인. 통각 과민은 중앙 통각 신경 세포 또는 중앙 탈 억제, 5, 6하지만 CPSP 증상에 관여하는 신경 기판을 알 수없는 남아의 hyperexcitability에 의해 발생할 수 있습니다. 일부 연구에서는 전전두엽 피질 (dPFC), 주동이의 앞쪽에 cingulate 피질 (ACC), 편도체, 해마, periaqueductal 그레이 (PAG), 주동이의 ventromedial 수질, 서로와의 연결이 침해 처리를 중재 할 것을 제안했다. 추가 7LY, 내측 전두엽 피질 (mPFC) -amygdala 회로는 CPSP의 병태 생리 학적 메커니즘에 8 데이터가 다양하다. 통증 관련 인식에 관여하는 것으로 도시 및 CPSP에 신경 기판의 활성화는 더 조사가 필요했다. [14 C] -Iodoantipyrine (IAP) 흡수는 뇌 활동 CBF의 관계를 가정하여, 간접적으로 지방 뇌 혈류 (rCBF)을 관찰하는 데 사용된다. 비록 [14 C] -IAP 이러한 기능 자기 공명 영상 (fMRI)와 같이, 실시간으로 뇌 활동을 평가할 수 없으며, 이는 여러 가지 장점을 갖는다. 예를 들어, [14 C] -IAP는 자발적으로 측정 병적 인 상태 동안 뇌 이벤트를 발생하기에 적합하다. (9) 또한, [14 C] -IAP 흡수가 마취없이 측정한다. 또한, 자기 공명과 양전자 방출 단층 촬영 (PET)를 포함하는 다른 이미징 방법보다 적은 비용이 든다. 에서 [14 C] -IAP 방법은 measurin에 적합한 것으로 제안되었다시상의 복부 기저 핵 (VB)의 병변에 의해 유도되는 g 자발 통증 (예를 들어, CPSP). (9) 본 프로토콜은 동물 모델에서 시상의 VB의 병변에 의해 유도되는 CPSP의 신경 기판의 참여를 평가하는 [14 C] -IAP 방법을 수행하는 방법에 대해 설명합니다. 이 기술은 행동과의 연결 수준 CPSP 증상을 기초 병리 생리 학적 메카니즘을 결정하는 방법을 제공한다.

<table border="0" cellpadding="0" cellspacing="0" width="785">
	<tbody>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:785px;">Anesthetic:</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Isoflurane</td>
			<td style="width:229px;">Halocarbon Products Corporation&#160;</td>
			<td style="width:173px;">NDC 12164-002-25</td>
			<td style="width:167px;">4%</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:785px;">Surgery</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">homeothermic blanket system</td>
			<td style="width:229px;">Harvard Apparatus</td>
			<td style="width:173px;">Model 50&#8211;7079</td>
			<td style="width:167px;">body temperature were maintained at 36.5-37.5&#176;C.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">10&#181;l micro syringe</td>
			<td style="width:229px;">Hamilton</td>
			<td style="width:173px;">80008, Model 1701SN</td>
			<td style="width:167px;">injected with collagenase</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">polyethylene-50 tubing</td>
			<td style="width:229px;">Becton, Dickinson and Company</td>
			<td style="width:173px;">427411</td>
			<td style="width:167px;">catheterized into external jugular vein</td>
		</tr>
		<tr height="45">
			<td height="45" style="height:45px;width:216px;">1 c.c syringe</td>
			<td style="width:229px;">Terumo Medical Products</td>
			<td style="width:173px;">SS-01T</td>
			<td style="width:167px;">injected with 14C-IAP and saline.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">saline (Sodium Chloride 0.9gm)</td>
			<td style="width:229px;">Taiwan Biotech Co., LTD.</td>
			<td style="width:173px;">100-130-0201</td>
			<td style="width:167px;">To flush the tube</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:785px;">Drugs:</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">type 4 collagenase</td>
			<td style="width:229px;">Sigma</td>
			<td style="width:173px;">C5138-500MG</td>
			<td style="width:167px;">0.125 U</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Gentamicin</td>
			<td style="width:229px;">Sigma</td>
			<td style="width:173px;">G1264-250MG</td>
			<td style="width:167px;">6 mg/kg</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heparin</td>
			<td style="width:229px;">Sigma</td>
			<td style="width:173px;">H9399</td>
			<td style="width:167px;">20 U/ml; 0.1 ml/day</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">14C-iodoantipyrine (IAP)</td>
			<td style="width:229px;">PerkinElmer</td>
			<td style="width:173px;">NEC712</td>
			<td style="width:167px;">125 mCi/kg in 300 ml of 0.9% saline</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Potassium chloride</td>
			<td style="width:229px;">Merck</td>
			<td style="width:173px;">1.04936.1000</td>
			<td style="width:167px;">3 M</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:785px;">Behavior system:</td>
		</tr>
		<tr height="126">
			<td height="126" style="height:126px;width:216px;">von Frey esthesiometer</td>
			<td style="width:229px;">Fabrication Enterprises, Inc.</td>
			<td style="width:173px;">Baseline Tactile Monofilaments 12-1666</td>
			<td style="width:167px;">mechanical hyperalgesia was assessed by measuring the withdrawal response to a mechanical stimulus</td>
		</tr>
		<tr height="126">
			<td height="126" style="height:126px;width:216px;">plantar test apparatus</td>
			<td style="width:229px;">IITC Life Science</td>
			<td style="width:173px;">IITC 390G Plantar Test</td>
			<td style="width:167px;">Thermal hyperalgesia was assessed by measuring the hind paw withdrawal latency in response to radiant heat.</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:785px;">Brain slice:</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Optimal Cutting Temperature compound</td>
			<td style="width:229px;">Sakura Fintek Inc</td>
			<td style="width:173px;">4583</td>
			<td style="width:167px;">embedded the brain</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">dry ice</td>
			<td style="width:229px;"></td>
			<td style="width:173px;"></td>
			<td style="width:167px;">frozen in dry ice/methylbutane (approximately &#8722;55&#176;C)</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">methylbutane</td>
			<td style="width:229px;">Sigma</td>
			<td style="width:173px;">M32631-1L</td>
			<td style="width:167px;">frozen in dry ice/methylbutane (approximately &#8722;55&#176;C)</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Cryostat</td>
			<td style="width:229px;">&#160;Leica Biosystems Nussloch GmbH, Germany</td>
			<td style="width:173px;">Leica CM1850</td>
			<td style="width:167px;">Coronal brain slice were sectioned on this machine.</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:785px;">Data analyze</td>
		</tr>
		<tr height="168">
			<td height="168" style="height:168px;width:216px;">exposure cassettes with a phosphor screen</td>
			<td style="width:229px;">Amersham Biosciences&#160;</td>
			<td style="width:173px;">20 cm x 25 cm</td>
			<td style="width:167px;">The slices were dried on glass slides and placed alongside five standard filter papers with graded radioactivity. All of the slides were exposed to the cassettes at &#8722;20&#176;C.</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">&#947;-counter</td>
			<td style="width:229px;">Beckman Coulter</td>
			<td style="width:173px;">Beckman LS 6500 Liquid Scintillation Counter</td>
			<td style="width:167px;">To measure the radioactivity count of the filter papers.</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Typhoon 9410 Variable Mode Imager</td>
			<td style="width:229px;">&#160;GMI, Inc.</td>
			<td style="width:173px;">WS-S9410</td>
			<td style="width:167px;">To read&#160; phosphor screen which was exposed by brain slice</td>
		</tr>
		<tr height="231">
			<td height="231" style="height:231px;width:216px;">Statistical Parametric Mapping (SPM)</td>
			<td style="width:229px;">Wellcome Centre for Neuroimaging</td>
			<td style="width:173px;">version 8</td>
			<td style="width:167px;">all of the brains were averaged to create the final brain template. To determine significant differences between the images in these two groups, the images were derived by subtracting the sham group from the CPSP group.</td>
		</tr>
		<tr height="294">
			<td height="294" style="height:294px;width:216px;">ImageJ</td>
			<td style="width:229px;">http://imagej.nih.gov/ij</td>
			<td style="width:173px;">version 1.46</td>
			<td style="width:167px;">Adjacent sections were aligned both manually and using Stack- Reg, an automated pixel-based registration algorithm in ImageJ software. All of the original three-dimensionally reconstructed brains were smoothed and normalized to the reference rat brain model.</td>
		</tr>
		<tr height="210">
			<td height="210" style="height:210px;width:216px;">Matlab</td>
			<td style="width:229px;">MathWorks</td>
			<td style="width:173px;">version 2009b</td>
			<td style="width:167px;">used Pearson correlation coefficients to examine the relationships between the CPSP and sham groups. An inter-regional correlation matrix was calculated across animals from each group.</td>
		</tr>
		<tr height="126">
			<td height="126" style="height:126px;width:216px;">Pajek</td>
			<td style="width:229px;">http://Pajek.imfm.si/</td>
			<td style="width:173px;">version 3.06</td>
			<td style="width:167px;">Graphical theoretical analysis was performed on networks defined by the above correlation matrices using Pajek software.</td>
		</tr>
	</tbody>
</table>

autoradiographic measurements of [14c]-iodoantipyrine in rat brain following central post-stroke pain

Approximately 8% of stroke patients present symptoms of central post-stroke pain (CPSP). CPSP is associated with allodynia and hypersensitivity to nociceptive stimuli. Although some studies have shown that neuropathic pain may involve the dorsolateral prefrontal cortex, rostral anterior cingulate cortex, amygdala, hippocampus, periaqueductal gray, rostral ventromedial medulla, and medial thalamus, the neural substrates and their connections that mediate CPSP remain unclear. [14C]-Iodoantipyrine (IAP) uptake can be measured to evaluate spontaneously active pain. It can be used to assess the activation of neural substrates that may be involved in CPSP in an animal model. The [14C]-IAP method in rats is less expensive to perform compared with other brain mapping techniques. The present [14C]-IAP protocol is used to measure the activation of neural substrates that are involved in CPSP that is induced by lesions of the ventral basal nucleus (VB) of the thalamus in a rodent model.

도 1a는 실험적인 타임 라인을 도시한다. 쥐가 행동 검사에 대한 사기 및 CPSP 그룹 (즉, 본 프레이 시험 및 발바닥 시험)에 할당되었다. 실험의 첫 날 기준으로 제공하고, 시험 1 주에 반복 - 4 헤파린 (20 U / ㎖, 0.1 ㎖ / 일) 동안 주입 일주일에 5 P​​E-50 카테터 외부 경정맥을 시행 하였다 주 (4)과 헤파린 주사 후 5. 5 분, [14 C] -IAP가 주입, 희생 마취?...

Watch this Scientific Journal Video about Autoradiographic Measurements of [14C]-Iodoantipyrine in Rat Brain Following Central Post-Stroke Pain at JoVE.com

Autoradiographic Measurements of [14C]-Iodoantipyrine in Rat Brain Following Central Post-Stroke Pain

We present a protocol to measure [14C]-iodoantipyrine (IAP) uptake and assess the activation of neural substrates that are involved in central post-stroke pain (CPSP) in a rodent model.

의 방사 법적 측정 [<sup&gt; (14)</sup&gt; 중앙 후 스트로크 통증에 따라 쥐의 뇌에서 C] -Iodoantipyrine

Approximately 8% of stroke patients present symptoms of central post-stroke pain (CPSP). CPSP is associated with allodynia and hypersensitivity to ...

autoradiographic-measurements-14c-iodoantipyrine-rat-brain-following

Research

JoVE Journal

Medicine

Autoradiographic Measurements of [14C]-Iodoantipyrine in Rat Brain Following Central Post-Stroke Pain

8.7K 조회수.  Fo Guang University. We present a protocol to measure [14C]-iodoantipyrine (IAP) uptake and assess the activation of neural substrates that are involved in central post-stroke pain (CPSP) in a rodent model.

비디오: Autoradiographic Measurements of [14C]-Iodoantipyrine in Rat Brain Following Central Post-Stroke Pain

In vitro Functional Characterization of Mouse Colorectal Afferent Endings

This video demonstrates in detail an in vitro single-fiber electrophysiological recording protocol using a mouse colorectum-nerve preparation. The approach allows unbiased identification and functional characterization of individual colorectal afferents. Extracellular recordings of propagated action potentials (APs) that originate from one or a few afferent (i.e., single-fiber) receptive fields (RFs) in the colorectum are made from teased nerve fiber fascicles. The colorectum is removed with either the pelvic (PN) or lumbar splanchnic (LSN) nerve attached and opened longitudinally. The tissue is placed in a recording chamber, pinned flat and perfused with oxygenated Krebs solution. Focal electrical stimulation is used to locate the colorectal afferent endings, which are further tested by three distinct mechanical stimuli (blunt probing, mucosal stroking and circumferential stretch) to functionally categorize the afferents into five mechanosensitive classes. Endings responding to none of these mechanical stimuli are categorized as mechanically-insensitive afferents (MIAs). Both mechanosensitive and MIAs can be assessed for sensitization (i.e., enhanced response, reduced threshold, and/or acquisition of mechanosensitivity) by localized exposure of RFs to chemicals (e.g., inflammatory soup (IS), capsaicin, adenosine triphosphate (ATP)). We describe the equipment and colorectum&#8211;nerve recording preparation, harvest of colorectum with attached PN or LSN, identification of RFs in the colorectum, single-fiber recording from nerve fascicles, and localized application of chemicals to the RF. In addition, challenges of the preparation and application of standardized mechanical stimulation are also discussed.

In vitro Functional Characterization of Mouse Colorectal Afferent Endings

This video demonstrates a protocol for conducting single-fiber electrophysiological recordings on an in vitro mouse colorectum-nerve preparation.

마우스 대장 수입 성의 엔딩의 생체 기능 특성에

This video demonstrates in detail an in vitro single-fiber electrophysiological recording protocol using a mouse colorectum-nerve preparation. The ...

연구

의학

The Monoiodoacetate Model of Osteoarthritis Pain in the Mouse

A major symptom of patients with osteoarthritis (OA) is pain that&#160;is triggered by peripheral as well as central changes within the pain pathways. The current treatments for OA pain such as NSAIDS or opiates are neither sufficiently effective nor devoid of detrimental side effects. Animal models of OA are being developed to improve our understanding of OA-related pain mechanisms and define novel pharmacological targets for therapy. Currently available models of OA in rodents include surgical and chemical interventions into one knee joint. The monoiodoacetate (MIA) model has become a standard for modelling joint disruption in OA in both rats and mice. The model, which is easier to perform in the rat, involves injection of MIA into a knee joint that induces rapid pain-like responses in the ipsilateral limb, the level of which can be controlled by injection of different doses. Intra-articular injection of MIA disrupts chondrocyte glycolysis by inhibiting glyceraldehyde-3-phosphatase dehydrogenase and results in chondrocyte death, neovascularization, subchondral bone necrosis and collapse, as well as inflammation. The morphological changes of the articular cartilage and bone disruption are reflective of some aspects of patient pathology. Along with joint damage, MIA injection induces referred mechanical sensitivity in the ipsilateral hind paw and weight bearing deficits that are measurable and quantifiable. These behavioral changes resemble some of the symptoms reported by the patient population, thereby validating the MIA injection in the knee as a useful and relevant pre-clinical model of OA pain.
The aim of this article is to describe the methodology of intra-articular injections of MIA and the behavioral recordings of the associated development of hypersensitivity with a mind to highlight the necessary steps to give consistent and reliable recordings.

Osteoarthritis (OA), or degenerative joint disease, is a debilitating condition associated with pain that remains only partially controlled by available analgesics. Animal models are being developed to improve our understanding of OA-related pain mechanisms. Here we describe the methodology for the monoiodoacetate model of OA pain in the mouse.

마우스에서 관절염 통증의 Monoiodoacetate 모델

A major symptom of patients with osteoarthritis (OA) is pain that&#160;is triggered by peripheral as well as central changes within the pain pathways. The ...

Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, we developed a rat model of patch angioplasty that can be used in either a vein or an artery, creating a patch venoplasty or a patch arterioplasty, respectively. Technical aspects of this model are discussed. The infra-renal IVC or aorta are dissected and then clamped proximally and distally. A 3 mm venotomy or arteriotomy is performed in the infrarenal inferior vena cava or aorta of 6 to 8 week-old Wistar rats. A bovine pericardial patch (3 mm x 1.5 mm x 0.6 mm) is then used to close the site using a 10-0 nylon suture. Compared to arterial patches, venous patches show increased neointimal thickness on postoperative day 7. This novel model of pericardial patch angioplasty can be used to examine neointimal hyperplasia on vascular biomaterials, as well as to compare the differences between the arterial and venous environments.

We have established a model of pericardial patch angioplasty that can be used in either small-diameter veins or arteries. This model can be used to compare venous and arterial neointimal hyperplasia formation.

쥐 대동맥 또는 하대 정맥에 혈관 성형술 패치

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, ...

A Model of Free Tissue Transfer: The Rat Epigastric Free Flap

Free tissue transfer has been increasingly used in clinical practice since the 1970s, allowing reconstruction of complex and otherwise untreatable defects resulting from tumor extirpation, trauma, infections, malformations or burns. Free flaps are particularly useful for reconstructing highly complex anatomical regions, like those of the head and neck, the hand, the foot and the perineum. Moreover, basic and translational research in the area of free tissue transfer is of great clinical potential. Notwithstanding, surgical trainees and researchers are frequently deterred from using microsurgical models of tissue transfer, due to lack of information regarding the technical aspects involved in the operative procedures. The aim of this paper is to present the steps required to transfer a fasciocutaneous epigastric free flap to the neck in the rat.
This flap is based on the superficial epigastric artery and vein, which originates from and drain into the femoral artery and vein, respectively. On average the caliber of the superficial epigastric vein is 0.6 to 0.8 mm, contrasting with the 0.3 to 0.5 mm of the superficial epigastric artery. Histologically, the flap is a composite block of tissues, containing skin (epidermis and dermis), a layer of fat tissue (panniculus adiposus), a layer of striated muscle (panniculus carnosus), and a layer of loose areolar tissue.
Succinctly, the epigastric flap is raised on its pedicle vessels that are then anastomosed to the external jugular vein and to the carotid artery on the ventral surface of the rat's neck. According to our experience, this model guarantees the complete survival of approximately 70 to 80% of epigastric flaps transferred to the neck region. The flap can be evaluated whenever needed by visual inspection. Hence, the authors believe this is a good experimental model for microsurgical research and training.

This paper describes the steps required to raise a fasciocutaneous epigastric free flap and transfer it to the neck in the rat.

무료 조직 전송의 모델 : 쥐 상복부 무료 플랩

Free tissue transfer has been increasingly used in clinical practice since the 1970s, allowing reconstruction of complex and otherwise untreatable defects ...

Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing and treating this complication using experimental models of spinal cord ischemia. The model described here demonstrates varying degrees of paraplegia that relate to the length of occlusion following thoracic aortic occlusion in a rat spinal cord ischemia model.
A 2-Fr. balloon-tipped catheter was advanced through the femoral artery into the descending thoracic aorta until the catheter tip was placed at the left subclavian artery in anesthetized male Sprague-Dawley rats. Spinal cord ischemia was induced by inflating the catheter balloon. After a set period of occlusion (9, 10, or 11 min), the balloon was deflated. Neurologic assessment was performed using the motor deficit index at 24 h after surgery, and the spinal cord was harvested for histopathological examination.
Rats that underwent 9 min of aortic occlusion showed mild and reversible motor impairment in the hind limb. Rats subjected to 10 min of aortic occlusion presented with moderate but reversible motor impairment. Rats subjected to 11 min of aortic occlusion displayed complete and persistent paralysis. The motor neurons in the spinal cord sections were more preserved in rats subjected to shorter duration of aortic occlusion.
Researchers can achieve a reproducible hind limb motor deficit following thoracic aortic occlusion using this spinal cord ischemia model.

This study demonstrates the technique to make a minimally invasive and easily reproducible model of spinal cord ischemia in rats. Various degrees of hind limb motor deficit can be produced by controlling the aortic occlusion time.

척수 쇄 허혈 모델에서 대동맥 폐쇄 후 재현성 운동 부족

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing ...

Spontaneous and Evoked Measures of Pain in Murine Models of Monoarticular Knee Pain

Pain is the main cause of disability from arthritis. There is currently an unmet need for adequate treatments for arthritis pain. Pre-clinical models are necessary and useful for studying the mechanisms of pain and for evaluating efficacy of arthritis therapies. Measuring pain in animal models of arthritis is challenging. We have developed methods for measuring evoked and spontaneous pain in three models of murine arthritis. We quantitate the evoked pain responses of mice subjected to firm palpation of a painful knee. We also evaluate spontaneous pain by the proportion of weight and the amount of time placed on each of their 4 limbs after induction of arthritis pain in one knee. Joint pain in these mouse models produces a significant increase in evoked pain responses and an alteration in weight bearing. Since mice are quadrupeds, they offload the painful limb to the contralateral limb, to the forelimbs, or some combination. These methods are simple, require minimal equipment, and are reproducible and sensitive for detecting pain. They are useful for studying both disease-modifying arthritis treatments and analgesics in mice.

We have developed an evoked measure of arthritis pain and coupled it with a standardized method for measuring spontaneous pain in different murine models of chemically induced arthritis. These measures are sensitive and reproducible for different types of joint pain.

Murine 모델 Monoarticular 무릎 통증의 통증의 자연과 갖는 측정

Pain is the main cause of disability from arthritis. There is currently an unmet need for adequate treatments for arthritis pain. Pre-clinical models are ...

A New Best Practice for Validating Tail Vein Injections in Rat with Near-infrared-Labeled Agents

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part of many experimental procedures where reagents need to be introduced directly into the bloodstream. Unwittingly, the injection can be subcutaneous, possibly altering the scientific outcomes. Utilizing a nanoemulsion-based biological probe with an incorporated near-infrared fluorescent (NIRF) dye, this method offers the capability of imaging a successful tail vein injection in vivo. With the use of a NIRF imager, images are taken before and after the injection of the agent. An acceptable IV injection is then qualitatively or quantitatively determined based on the intensity of the NIRF signal at the site of injection.

Here we present a method to validate tail vein injections in rats by utilizing near-infrared fluorescence imaging data from dyes incorporated into agents or biological probes. The tail is imaged before and after the injection, the fluorescent signal is quantified, and an assessment of the injection quality is made.

근처-적외선-표시 에이전트와 쥐의 꼬리 정 맥 주사의 유효성 검사에 대 한 새로운 좋습니다

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part ...

Blood Collection Through Subclavian Vein Puncture in Mice

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in research work. Tail blood collection is a popular method when a small amount of blood sample is needed. Orbital artery could be considered if a large amount of blood is needed but this blood collection method has ethical issues. Formerly, we demonstrated the feasibility and safety of blood sample collection through subclavian vein puncture in rats, and here we investigate whether this method could be used in mice. We report that this method is safe and practical for blood collection in mice. Blood collection through the subclavian vein puncture in mice can be a convenient method in daily research works.

Here, we present a protocol to take blood samples from the subclavian vein of mice.

생쥐의 쇄 골 정 맥 천자를 통한 혈액 채취

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in ...

Functional and Physiological Methods of Evaluating Median Nerve Regeneration in the Rat

The main goal of this investigation is to show how to create and repair different types of median nerve (MN) lesions in the rat. Moreover, different methods of simulating postoperative physiotherapy are presented. Multiple standardized strategies are used to assess motor and sensory recovery using an MN model of peripheral nerve lesion and repair, thus permitting easy comparison of the results. Several options are included for providing a postoperative physiotherapy-like environment to rats that have undergone MN injuries. Finally, the paper provides a method to evaluate the recovery of the MN using several noninvasive tests (i.e., grasping test, pin prick test, ladder rung walking test, rope climbing test, and walking track analysis), and physiological measurements (infrared thermography, electroneuromyography, flexion strength evaluation, and flexor carpi radialis muscle weight determination). Hence, this model seems particularly appropriate to replicate a clinical scenario, facilitating extrapolation of results to the human species.
Although the sciatic nerve is the most studied nerve in peripheral nerve research, analysis of the rat MN presents various advantages. For example, there is a reduced incidence of joint contractures and automutilation of the affected limb in MN lesion studies. Furthermore, the MN is not covered by muscle masses, making its dissection easier than that of the sciatic nerve. In addition, MN recovery is observed sooner, because the MN is shorter than the sciatic nerve. Also, the MN has a parallel path to the ulnar nerve in the arm. Hence, the ulnar nerve can be easily used as the nerve graft for repairing MN injuries. Finally, the MN in rats is located in the forelimb, akin to the human upper limb; in humans, the upper limb is the site of most peripheral nerve lesions.

Presented is a protocol to produce different types of median nerve (MN) lesions and repair in the rat. Additionally, the protocol shows how to evaluate the functional recovery of the nerve using several noninvasive behavioral tests and physiological measurements.

쥐에서 중간 신경 재생을 평가하는 기능적 및 생리적 방법

The main goal of this investigation is to show how to create and repair different types of median nerve (MN) lesions in the rat. Moreover, different ...

Protocol for Developing a Femur Osteotomy Model in Wistar Albino Rats

Fracture healing is a physiological process resulting in the regeneration of bone defects by the coordinated action of osteoblasts and osteoclasts. Osteoanabolic drugs have the potential to augment the repair of fractures but have constraints like high costs or undesirable side effects. The bone healing potential of a drug can initially be determined by in vitro studies, but in vivo studies are needed for the final proof of concept. Our objective was to develop a femur osteotomy rodent model that could help researchers understand the development of callus formation following fracture of the shaft of the femur and that could help establish whether a potential drug has bone healing properties. Adult male Wistar albino rats were used after Institutional Animal Ethics Committee clearance. The rodents were anesthetized, and under aseptic conditions, complete transverse fractures at the middle one-third of the shafts of the femurs were created using open osteotomy. The fractures were reduced and internally fixed using intramedullary K-wires, and secondary fracture healing was allowed to take place. After surgery, intraperitoneal analgesics and antibiotics were given for 5 days. Sequential weekly x-rays assessed callus formation. The rats were sacrificed based on radiologically pre-determined time points, and the development of the fracture callus was analyzed radiologically and using immunohistochemistry.

Here, we present a protocol to iatrogenically fracture the shaft of the femur of Wistar albino rats and follow up on the development of the callus. This femur osteotomy model can help researchers evaluate the process of fracture healing and to study how a drug could influence fracture healing.

Wistar 알비노 쥐에서 대퇴골 절골술 모델을 개발하기 위한 프로토콜

Fracture healing is a physiological process resulting in the regeneration of bone defects by the coordinated action of osteoblasts and osteoclasts. ...

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