저자는 베넷과 샤오 (1988)에 의해 CCI 모델의 원래 설명을 인정하고 싶습니다. 이 작품은 과학 그리니치 표준시에 의학 연구를위한 국민 건강과 호주의 의료 연구 협의회 (ID # 568637)과 뉴사우스 웨일즈 사무소에서 교부금에 의해 부분적으로 지원되었다.

1. 좌골 신경 만성 수축 무균 기술은 수술을 위해 사용해야합니다. 70 % 에탄올로 수술 작업 표면을 소독하고 autoclaving에 의해 사전에 멸균 악기 (예 : 미세 가위, 포셉, 그리고 견인기), gauzes, 스테이플과 면봉으로 준비합니다. 여러 수술의 경우 청소 및 70 % 에탄올이나 쥐 사이에 마른 비드 steriliser와 resterilise 악기. 외과 마스크, 헤어 모자와 멸균 장갑을 착용한다. 신경의 수축 들어, 건조 방지를 위해, 멸균 식염수에 포장되어 약 3 cm 길이의 작은 조각으로 절단하여 삼가의 크롬을 함유하는 장치 내장 봉합사를 준비합니다. 상처를 닫을 경우, 상용 멸균 실크 봉합을 사용합니다. <ol><li> O 2의 isoflurane 5 %를 사용하는 유도 챔버에 쥐를 Anaesthetise 다음, 안면 만든 사용자 정의를 통해 O 2에서 isoflurane 2 % 제공하고, 절차를 수행하는 동안 anesthetised 동물을 모니터링. 일반적인 대안 anesthetics는 할로탄 또는 intraperitoneal 나트륨은 pentobarbital 흡입됩니다. 이들 수용체는 neuropathic 고통의 개발에 중요한 이후 케타민을은 N-메틸-D-aspartate (NMDA) 수용체 길항제로 피해야한다. </li><li> 쥐의 왼쪽 뒷다리 다리를 면도. 그런 다음, 37에서 열 조절 난방 매트에 동물을 배치 ° C와 눈에 윤활 안과 연고를 적용합니다. </li><li> 70% 이소 프로필 알코올과 요오드 용액 세 가지 다른 응용 프로그램과 함께 면도 영역을 소독. </li><li> 쥐가 가슴 / 흉부에 누워으로 왼쪽 뒷다리 다리를 위로 향하게하고 발을 마스킹 테이프를 사용하여 척추 90 °에서 femor과 위치에 누르고 있습니다. </li><li> 피부 병렬 절개하지만 femor 아래 3~4mm, 그리고 결합 조직을 절단하여 절개를 둘러싼 근육에서 무료로 피부를 확인합니다. </li><li> gluteus superficialis와 팔뚝 사이의 결합 조직을 잘라 무딘 가위를 이용하여femoris 근육. </li><li> 좌골 신경의 명확한 시각화 수 있도록,이 두 근육 사이의 간격을 넓혀 리트 렉터를 사용합니다. </li><li> 곡선 야기한 스쳐 포셉 및 마이크로 가위, 주변의 결합 조직에서 좌골 신경의 부드럽게 무료로 약 10 밀리 (좌골 trifurcation에 근위)를 사용. </li><li> 현미경과 좋은 광원에서, 이중 매듭으로 좌골 신경의 trifurcation에 근위 간격 1mm를, 넷 ligatures을 (삼가의 크롬을 함유하는 직감 4.0, Ethicon, 미국) 매다. 각각의 끈 들어, 하나의 느슨한 루프로 시작, 다음 루프 가까운 두 끝을 파악하고 루프 살짝만 아늑한되고 봉합사는 신경을 따라 미끄러지게하지 않는까지 조입니다. 의 적절한 위치에 루프를 개최하려면, 매듭을 완성을 가장 먼저 위에 두 번째 루프를 놓으십시오. 마지막으로, 1mm 주위에 묶인의 미심쩍은 부분을 절단. 신경의 수축이 최소화되고, 간단한 떨리는이 관찰되면 즉시 중지, 일 때문에 체포되지 않도록해야전자 epineural 혈액의 흐름. ligatures 단단히 걸쳐 axotomy로 연결하고, autotomy (자동 절단), 성공적인 통증 과민성 테스트를 배제 모두 원치 않는 부작용. </li><li> 피부를 고정하는 근육 레이어와 스테이플 (Autoclip, 9 밀리) 닫습 봉합 (Mersilk 5.0, Ethicon)를 사용합니다. 다음, 상처를 소독하기 (Riodine) 요오드 용액을 사용하십시오. </li><li> 질식 의식 동물을 방지하기 위해, 마취 회복 기간 동안 긴밀하게 쥐를 관찰하고, 그것이 평평한 종이 침구 (아니라 표준 동물 축산의 침구)와 별도의 새장에 복구할 수 있습니다. 어떤 수술 진통제의 수사를 제공하기 전에 이것이 통증 과민성 시험을 방해할 수있는 사실을 고려해야합니다. 사실, 부상 당시 마취제, lidocaine의 로컬 응용 프로그램이 neuropathic 고통의 발전을 변경하고, 이전 주 7 hyperalgesia의 정도를 감소 수있다는 증거가있다. 이 m에 대한 논의는베넷 외 8 시까지 리뷰를보고 atter. </li></ol> 2. 통증 Behaviours의 측정 <ol><li> 실험을 시작하기 전에 며칠 동안 쥐의 일일 처리는 해당 처리기에 쥐가 의히다 스트레스와 물고의 위험을 줄이기 위해 권장합니다. </li><li> 행동적 테스트에 앞서, 쥐가 테스트 절차 및 동물 인클로저에게 길들여해야합니다. 테스트 환경은 조용하고 보관하고 잘 일정한 온도 및 습도 수준으로 조절하고, 하루 중 비슷한 시간을 각 테스트 세션에서 수행되어야합니다. </li><li> 통증 과민성 테스트는 원하는대로, CCI (기준) 이전과 CCI 후 다른 시간 지점에서 여러 번 수행해야합니다. </li></ol> 3. 기계 탈퇴 임계값 <ol><li> 시험 당일, 장소의 쥐 고가 메쉬 낭패 테스트 케이지로 탈퇴 임계값을 측정하기 전에 15-30분, 새장 탐사 및 groo있게명나라 활동을 중단하고 쥐가 편안하고 고정이 될 수 있습니다. </li><li> 직접 hindpaw의 중반 발바닥 표면 바로 아래에 동적 발바닥 폰 프라이 aesthesiometer (우고 Basile, 이탈리아), 위치 필라멘트와 터치 자극기 장치를 사용하여 금속 메쉬 피하 시작 키를 누르십시오. 이것은 기계적으로 증가 힘으로 hindpaw의 발바닥 표면을 자극하기 위해 필라멘트를 올렸으니까요. </li></ol> 수동 폰 프라이의 머리카락은 피부에 적용할 때 특정 무력을 유도하도록 조정되는 직경 필라멘트 증가의 연속입니다. 그들의 발자국이 인출 한도 등 이전 4 설명된 측정을위한 전자 동적 발바닥 폰 프라이의 aesthesiometer의 대안으로 사용될 수 있습니다. 전자 폰 프라이 aesthesiometers (예 IITC, 미국 및 Somedic, 스웨덴)의 여러 다른 제조 업체도 있습니다. <ol start="3"><li> 부상 및 손상되지 않은 두 hindpa에서 기계 탈퇴 임계값을 기록WS, counterbalanced 순서를 유지해야합니다. 이 장치는 웃기고 압력을 적용하고 금단 반응에 따라 시스템이 자동으로 기록하고 (g)을 적용 최강 군사력을 표시합니다. 모호한 응답 (ambulation이나 정리의 경우 IE)가 발생하면, 자극을 반복하고 다시 임계값을 기록합니다. 빠는데 또는 hindpaw의 다음의 자극을 떨고있는 것은 고통과 관련된 행동의 징후입니다. </li><li> 기계적 자극에게 자극 사이에 약 5 분 간격으로 3 회 반복 임계값을 기록하고 발자국 탈퇴 임계값의 평균을 계산합니다. </li></ol> 4. 열 탈퇴 대기 시간 발바닥 테스트 악기의 적외선 (IR​​) 가열 강도는 실험을 시작하기 전에 조정해야합니다. 저희 연구실에서는, 우리는 순진 쥐 10 초 정도의 평균 발자국 탈퇴 대기 시간을 제공하기 위해 적외선 강도를 조절, 우리는 조직 댐을 피하기 데는 20 초 컷 - 오프 지연 시간을 설정앞발을 제거하는 실패의 경우 연령. <ol><li> 시험 당일, 장소 15-30분 지연을 측정하기 전에 유리 낭패 테스트 케이지에 쥐, 케이지 탐사를 허용하고 중단을위한 활동을 정리하고 쥐가 편안하고 고정이되었습니다. </li><li> 직접 hindpaw의 중반 발바닥 표면 바로 아래에 열 발바닥 진통제 악기 (우고 Basile, 이탈리아), 위치 적외선 소스를 사용하여 시작 키를 누르십시오. 이것은 투명한 유리 표면을 통해 복사열의 광선에 hindpaw의 발바닥 표면을 제공합니다. 열 발바닥 진통제 악기 (예 : IITC, 미국 및 Stoelting, 미국)의 다른 제조 업체도 있습니다. </li><li> counterbalanced 위해 부상과 손상되지 않은 두 hindpaws에서 탈퇴 대기 시간을 기록합니다. 이 장치는 자동으로 열원에서 손 철회에 대한 열 자극의 발병에서 (초) 촬영 시간을 기록합니다. 모호한 반응이 발생하는 경우 (ambula의 경우 즉,기 또는 정리), 자극을 반복하고 다시 대기 시간을 기록합니다. 빠는데 또는 hindpaw의 다음의 자극을 떨고있는 것은 고통과 관련된 행동의 징후입니다. </li><li> 열 자극에게 자극 사이에 약 5 분 간격으로 적어도 3 번 반복, 대기 시간을 기록하고 발자국 탈퇴 대기 시간의 평균을 계산합니다. </li></ol> 4. 대표 결과 Wistar 쥐의 (손상되지 않은) ipsilateral (부상)과 contralateral 발냄새의 기계적 인출 한도 및 열 회수 지연의 예제는 앞에 표시 (베이스 라인) 및 2~12일 그림 3A의 CCI, 그리고 그림 3B 이후. 있습니다 이러한 수치에서 그것은 CCI는, 기계적 및 열적 통증 과민성의 존재를 나타내는, 기계 금단 임계값과 ipsilateral hindpaw의 열 인출 대기 모두에서 상당한 감소를 일으키는 것을 볼 수있는 만성 neuropathic 고통의 두 기능<html> . <img alt="그림 1" src="/files/ftp_upload/3393/3393fig1.jpg" /> 그림 1. CCI 모델 ligatures의 위치를 보여주는 개략도. neuropathic 고통의이 동물 모델은 중간 허벅지 수준에서 좌골 신경 네개의 느슨한 삼가의 크롬을 함유하는 - 내장 ligatures의 배치를 포함한다. 그것은 hindpaw 기슭와 후속 통증 과민성에가는 afferents의 일부만 손상에 대한 결과입니다. <img alt="그림 2" src="/files/ftp_upload/3393/3393fig2.jpg" /> 그림 2. 통증이 테스트에 사용 hindpaw 영역의 다이어그램. 같은 빨간색 점선 동그라미로 표시 footpads에서 불과 후부 중반 발바닥 표면은, 고통 테스트에 사용됩니다. 이 지역은 좌골 신경의 말단 지점에서 주로 innervated 있습니다. 뒤꿈치와 footpads는 차동 피부 민감도에 따라 피해야한다. &quot;3.jpg /&gt; 쥐의 좌골 신경의 만성적인 수축에 따라 그림 3. 통증 과민성. CCI 후, 기계적 자극 (A)와 (B) 부상에서 열 자극에 발자국 탈퇴 대기 시간 (ipsilateral)에 발자국 탈퇴 임계값에 상당한 감소는 손상되지 않은 (contralateral) hindpaw (P는 &lt;0.01 반복 측정 2로 비교할 hindpaw입니다 방향 ANOVA). 데이터는 뜻 보여주 ± SEM, n은 = 7 Wistar의 쥐.

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R., Bandler, R., Keay, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+subpopulation+of+rats+show+social+and+sleep-waking+changes+typical+of+chronic+neuropathic+pain+following+peripheral+nerve+injury.">A subpopulation of rats show social and sleep-waking changes typical of chronic neuropathic pain following peripheral nerve injury.</a> Eur. J. Neurosci. 17, 1907-1920 (2003).</li><li>Roeska, K., Doods, H., Arndt, K., Treede, R. D., Ceci, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anxiety-like+behaviour+in+rats+with+mononeuropathy+is+reduced+by+the+analgesic+drugs+morphine+and+gabapentin.">Anxiety-like behaviour in rats with mononeuropathy is reduced by the analgesic drugs morphine and gabapentin.</a> Pain. 139, 349-357 (2008).</li><li>Kontinen, V. K., Kauppila, T., Paananen, S., Pertovaara, A., Kalso, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+measures+of+depression+and+anxiety+in+rats+with+spinal+nerve+ligation-induced+neuropathy.">Behavioural measures of depression and anxiety in rats with spinal nerve ligation-induced neuropathy.</a> Pain. 80, 341-346 (1999).</li><li>Urban, R., Scherrer, G., Goulding, E. H., Tecott, L. H., Basbaum, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+indices+of+ongoing+pain+are+largely+unchanged+in+male+mice+with+tissue+or+nerve+injury-induced+mechanical+hypersensitivity.">Behavioral indices of ongoing pain are largely unchanged in male mice with tissue or nerve injury-induced mechanical hypersensitivity.</a> Pain. 152, 990-1000 (2011).</li><li>Myers, R. R., Yamamoto, T., Yaksh, T. L., Powell, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+focal+nerve+ischemia+and+Wallerian+degeneration+in+peripheral+nerve+injury+producing+hyperesthesia.">The role of focal nerve ischemia and Wallerian degeneration in peripheral nerve injury producing hyperesthesia.</a> Anesthesiology. 78, 308-316 (1993).</li><li>Grace, P. M., Hutchinson, M. R., Manavis, J., Somogyi, A. A., Rolan, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+animal+model+of+graded+neuropathic+pain:+utility+to+investigate+mechanisms+of+population+heterogeneity.">A novel animal model of graded neuropathic pain: utility to investigate mechanisms of population heterogeneity.</a> J. Neurosci. Methods. 193, 47-53 (2010).</li><li>Kajander, K. C., Pollock, C. H., Berg, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+hindpaw+position+in+rats+during+chronic+constriction+injury+&#40;CCI&#41;+produced+with+different+suture+materials.">Evaluation of hindpaw position in rats during chronic constriction injury &#40;CCI&#41; produced with different suture materials.</a> Somatosens Mot. Res. 13, 95-101 (1996).</li><li>Xu, J., Pollock, C. H., Kajander, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chromic+gut+suture+reduces+calcitonin-gene-related+peptide+and+substance+P+levels+in+the+spinal+cord+following+chronic+constriction+injury+in+the+rat.">Chromic gut suture reduces calcitonin-gene-related peptide and substance P levels in the spinal cord following chronic constriction injury in the rat.</a> Pain. 64, 503-509 (1996).</li><li>Sommer, C., Schafers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Painful+mononeuropathy+in+C57BL/Wld+mice+with+delayed+wallerian+degeneration:+differential+effects+of+cytokine+production+and+nerve+regeneration+on+thermal+and+mechanical+hypersensitivity.">Painful mononeuropathy in C57BL/Wld mice with delayed wallerian degeneration: differential effects of cytokine production and nerve regeneration on thermal and mechanical hypersensitivity.</a> Brain. Res. 784, 154-162 (1998).</li><li>Walczak, J. S., Beaulieu, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+three+models+of+neuropathic+pain+in+mice+using+a+new+method+to+assess+cold+allodynia:+the+double+plate+technique.">Comparison of three models of neuropathic pain in mice using a new method to assess cold allodynia: the double plate technique.</a> Neurosci. Lett. 399, 240-244 (2006).</li><li>Maves, T. J., Pechman, P. S., Gebhart, G. F., Meller, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Possible+chemical+contribution+from+chromic+gut+sutures+produces+disorders+of+pain+sensation+like+those+seen+in+man.">Possible chemical contribution from chromic gut sutures produces disorders of pain sensation like those seen in man.</a> Pain. 54, 57-69 (1993).</li><li>Tall, J. M., Stuesse, S. L., Cruce, W. L., Crisp, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+and+the+behavioral+manifestations+of+neuropathic+pain.">Gender and the behavioral manifestations of neuropathic pain.</a> Pharmacol Biochem. Behav. 68, 99-104 (2001).</li><li>Chung, J. M., Choi, Y., Yoon, Y. W., Na, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+age+on+behavioral+signs+of+neuropathic+pain+in+an+experimental+rat+model.">Effects of age on behavioral signs of neuropathic pain in an experimental rat model.</a> Neurosci. Lett. 183, 54-57 (1995).</li><li>Tanck, E. N., Kroin, J. S., McCarthy, R. J., Penn, R. D., Ivankovich, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+age+and+size+on+development+of+allodynia+in+a+chronic+pain+model+produced+by+sciatic+nerve+ligation+in+rats.">Effects of age and size on development of allodynia in a chronic pain model produced by sciatic nerve ligation in rats.</a> Pain. 51, 313-316 (1992).</li><li>Shir, Y., Sheth, R., Campbell, J. N., Raja, S. N., Seltzer, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soy-containing+diet+suppresses+chronic+neuropathic+sensory+disorders+in+rats.">Soy-containing diet suppresses chronic neuropathic sensory disorders in rats.</a> Anesth. Analg. 92, 1029-1034 (2001).</li><li>Yoon, Y. W., Lee, D. H., Lee, B. H., Chung, K., Chung, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Different+strains+and+substrains+of+rats+show+different+levels+of+neuropathic+pain+behaviors.">Different strains and substrains of rats show different levels of neuropathic pain behaviors.</a> Experimental Brain Research. 129, 167-171 (1999).</li><li>Mogil, J. S., Davis, K. D., Derbyshire, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+necessity+of+animal+models+in+pain+research.">The necessity of animal models in pain research.</a> Pain. 151, 12-17 (2010).</li><li>Mogil, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+pain:+progress+and+challenges.">Animal models of pain: progress and challenges.</a> Nat. Rev. Neurosci. 10, 283-294 (2009).</li><li>Langford, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coding+of+facial+expressions+of+pain+in+the+laboratory+mouse.">Coding of facial expressions of pain in the laboratory mouse.</a> Nat. Methods. 7, 447-449 (2010).</li><li>Austin, P. J., Moalem-Taylor, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neuro-immune+balance+in+neuropathic+pain:+involvement+of+inflammatory+immune+cells,+immune-like+glial+cells+and+cytokines.">The neuro-immune balance in neuropathic pain: involvement of inflammatory immune cells, immune-like glial cells and cytokines.</a> J. Neuroimmunol. 229, 26-50 (2010).</li></ol>

동물 실험은 뉴 사우스 웨일즈, 오스트 레일 리아 대학의 동물 관리 및 윤리위원회의 승인을, 그리고 비탄의 연구를위한 국제 교류 협회에서 발행한 지침을 따라했다.

CCI는 만성 neuropathic 고통의 널리 사용되는 말초 신경 손상 모델입니다. 그것은 수행 비교적 간단하고, 부상 후 최소 한 달 동안 강력하고 안정적​​인 통증 과민성을 생산하고 있습니다. CCI 따라 쥐가 부상 hindpaw (발가락이 함께 개최되어 발바닥 - 근육이 수축과 앞발로 땅을 차다는 everted)뿐만 아니라, 흔들림 지키고 자발적인 고통 9 존재를 암시 부상 hindpaw으로 핥아 반복 비정상적인 자?...

<table border="1"><tbody><tr><td> 시약 / 장비의 명칭 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 삼가의 크롬을 함유하는 배 </td><td> Ethicon </td><td> G-211 </td><td> 4-0 두께 </td></tr><tr><td> Isoflurane </td><td> Delvet Pty. (주), 세븐 힐즈, 뉴사우스 웨일즈 (NSW) </td><td> N / A </td><td></td></tr><tr><td> Mersilk </td><td> Ethicon </td><td> W-580 </td><td> 5-0 두께 </td></tr><tr><td> Autoclip </td><td> Becton 디킨슨 </td><td> 427,631 </td><td> 9mm 스테인리스 </td></tr><tr><td> Riodine </td><td> 오리온 </td><td> R1000802F </td><td> 1퍼센트 W / V 요오드 </td></tr><tr><td> 열 발바닥 진통제 악기 </td><td> 우고 Basile </td><td> 37,370 </td><td></td></tr><tr><td> 동적 발바닥 aesthesiometer </td><td> 우고 바질전자 </td><td> 37,400 </td><td></td></tr></tbody></table>

chronic constriction of the sciatic nerve and pain hypersensitivity testing in rats

만성 neuropathic 고통은 중앙 또는 주변 신경계 손상으로 인한, 인구 1,2의 7~18% 영향을 만연하고 쇠약 조건이다. 증상은 자연 (같은 간지, 굽기, 전기 충격) 고통, dysaesthesia, paraesthesia, allodynia (통증이 보통이 아닌 고통스러운 자극으로 인한) 및 hyperalgesia합니다 (통증 자극에 대한 향상된 응답)가 있습니다. 감각 증상은 불면증과 우울증과 같은 행동 장애와 공동 병적입니다. 만성 neuropathic 고통을 공부하고 말초 신경 손상을 흉내낸 여러 동물 모델이 개발되었고, 가장 널리 사용되는 중 하나는 베넷과 샤오의 (1988) 일방적 좌골 신경 만성 수축 상해 (CCI) 3 (그림 1)입니다. 여기 CCI 및 테스트 고통 과민성을 수행하기위한 방법을 제시한다. CCI는 다음과 같이함으로써 노출된 한쪽에서 좌골 신경으로 마취하에 수행됩니다친척 절개하고 gluteus superficialis와 팔뚝 femoris 근육 사이의 결합 조직을 통해 절단. 네 삼가의 크롬을 함유하는 제품 내장 ligatures 그냥 윗니와 아랫니가 맞물리다 위해, 1 밀리미터 간격으로 좌골 신경 주위에 느슨하게 연결되어 있지만 epineural 혈류를 체포되지 않습니다. 상처가 피부의 근육 및 스테이플의 봉합으로 닫힙니다. 동물 그때 통증 과민성 시험이 시작되기 전에 24 시간 동안 수술에서 회복하기 위해 허용됩니다. 행동 테스트를 위해 쥐나는 테스트 장치에 배치하고 테스트 절차에 길들 수있게됩니다. 테스트 영역은 좌골 신경 분포에 속하는 hindpaw의 중반 발바닥 표면 (그림 2)이다. 기계 인출 한도는 기계 전자 동적 발바닥 폰 프라이 aesthesiometer 또는 수동 폰 프라이 머리카락 4를 사용하여 부상 및 손상되지 않은 hindpaws을 모두 자극에 의해 평가된다. 기계 인출 한도는 진력 최대 압력(g)를 그것의 발자국이 탈퇴를 실행시킵니다. 첫째 학스다 외 (1988)에 기술된 열 인출 지연의 측정은 hindpaw는 발바닥 진통제 측정기 5,6를 사용하여 투명한 유리 표면을 통해 복사열의 광선에 노출됩니다. 열 자극에 대한 탈퇴 대기 시간은 부상과 손상되지 않은 hindpaws 모두에서 발자국 철회를위한 시간으로 기록됩니다. 이어 CCI, 부상 한것 땜에 기계적 인출 한도뿐만 아니라 열 인출 지연 시간 기준 측정과 손상되지 않은 앞발로 땅을 차다 (그림 3)에 비해 모두 크게 줄일 수 있습니다. 통증 과민성 테스트와 함께 말초 신경 손상의 CCI 모델은 만성 neuropathic 고통을 수정하는 잠재적인 치료 대리인의 효과를 조사하기위한 모델 시스템을 제공합니다. 저희 연구실에서는 pathogenesis에 신경 면역 상호 작용의 역할을 조사하기 위해 hindpaws의 열 및 기계적 감도와 함께 CCI를 활용neuropathic 고통과 치료.

Watch this Scientific Journal Video about Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats at JoVE.com

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

수술의 단순하고 강력한 행동 결과로 인해 좌골 신경의 만성적인 수축이 neuropathic 고통의 사전 저명한 동물 모델 중 하나입니다. 수술에 따라 24 시간 내에 통증 과민성 설립되고 양적 폰 프라이 aesthesiometer (기계적 시험)과 발바닥 진통제 측정기 (온도 시험)를 사용하여 측정할 수있다.

좌골 신경 및 쥐의 통증 과민성 시험의 만성 수축

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

chronic-constriction-sciatic-nerve-pain-hypersensitivity-testing

Research

JoVE Journal

Medicine

58.6K 조회수.  University of New South Wales. 수술의 단순하고 강력한 행동 결과로 인해 좌골 신경의 만성적인 수축이 neuropathic 고통의 사전 저명한 동물 모델 중 하나입니다. 수술에 따라 24 시간 내에 통증 과민성 설립되고 양적 폰 프라이 aesthesiometer (기계적 시험)과 발바닥 진통제 측정기 (온도 시험)를 사용하여 측정할 수있다.

비디오: Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

쥐의 척수를 다친 후에 급성 및 만성 촉각 감각 테스트

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

연구

의학

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart failure. Here, we describe a detailed surgical procedure for creating pressure overload and cardiac hypertrophy in rats by constriction of the ascending aorta using a small metallic clip. After anesthesia, the trachea is intubated by inserting a cannula through a half way incision made between two cartilage rings of trachea. Then a skin incision is made at the level of the second intercostal space on the left chest wall and muscle layers are cleared to locate the ascending portion of aorta. The ascending aorta is constricted to 50&#8211;60% of its original diameter by application of a small sized titanium clip. Following aortic constriction, the second and third ribs are approximated with prolene sutures. The tracheal cannula is removed once spontaneous breathing was re-established. The animal is allowed to recover on the heating pad by gradually lowering anesthesia. The intensity of pressure overload created by constriction of the ascending aorta is determined by recording the pressure gradient using trans-thoracic two dimensional Doppler-echocardiography. Overall this protocol is useful to study the remodeling events and contractile properties of the heart during the gradual onset and progression from compensated cardiac hypertrophy to heart failure stage.

We describe a stepwise procedure for creating pressure overload and left ventricular hypertrophy in Wistar rats by constriction of the ascending aorta using a small metallic clip. This model is extensively used for studying remodeling changes during cardiac hypertrophy and for identifying and evaluating strategies for regression of such changes.

압력 과부하 심장 비대 모델의 창조를위한 쥐의 대동맥 수축 오름차순

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

쥐의 방광 통증 모델로 방광 팽창 유발 내장 운동 응답

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

The Sciatic Nerve Cuffing Model of Neuropathic Pain in Mice

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in injured sensory neurons and along the entire nociceptive pathway within the central nervous system. It is usually chronic and challenging to treat. In order to study neuropathic pain and its treatments, different models have been developed in rodents. These models derive from known etiologies, thus reproducing peripheral nerve injuries, central injuries, and metabolic-, infectious- or chemotherapy-related neuropathies. Murine models of peripheral nerve injury often target the sciatic nerve which is easy to access and allows nociceptive tests on the hind paw. These models rely on a compression and/or a section. Here, the detailed surgery procedure for the &#34;cuff model&#34; of neuropathic pain in mice is described. In this model, a cuff of PE-20 polyethylene tubing of standardized length (2 mm) is unilaterally implanted around the main branch of the sciatic nerve. It induces a long-lasting mechanical allodynia, i.e., a nociceptive response to a normally non-nociceptive stimulus that can be evaluated by using von Frey filaments. Besides the detailed surgery and testing procedures, the interest of this model for the study of neuropathic pain mechanism, for the study of neuropathic pain sensory and anxiodepressive aspects, and for the study of neuropathic pain treatments are also discussed.

Neuropathic pain is a consequence of a lesion or disease affecting the somatosensory system. The &#8220;cuff model&#8221; of neuropathic pain in mice consists of the implantation of a polyethylene cuff around the main branch of the sciatic nerve. Mechanical allodynia is tested using von Frey filaments.

마우스의 신경 병증 성 통증의 좌골 신경의 바지를 접 모델

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in ...

Chronic Constriction Injury of the Rat's Infraorbital Nerve (IoN-CCI) to Study Trigeminal Neuropathic Pain

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral procedures to study trigeminal neuropathic pain in rats. To meet the specificity of trigeminal neuropathic pain syndromes, the infraorbital nerve (IoN) is subjected to a chronic constriction injury (CCI) by loosely ligating the nerve. An intra-orbital approach is presented here to expose and ligate the IoN in the orbital cavity. After IoN ligation, rats exhibit changes in spontaneous behavior and in response to von Frey hair stimulation that are indicative of persistent pain and mechanical allodynia. Two phases can be defined in the development of the behavioral changes. During the first week following IoN-CCI (phase 1), rats show an increased and asymmetric face grooming activity, i.e., with face wash strokes primarily directed to the nerve-injured IoN territory. A distinction is made between face grooming behavior that is part of a more general body grooming behavior, which remains largely unaffected by IoN-CCI, and face grooming that is neither preceded nor followed by body grooming, which is significantly increased after IoN-CCI. During this period, responsiveness to mechanical stimulation of the IoN territory is reduced. This hyporesponsiveness is abruptly replaced by an extreme hyperresponsiveness whereby even very weak stimulus intensities provoke nocifensive behavior (phase 2). The phenomenological similarities between these behavioral alterations and reported signs of facial pain (i.e., responses to noxious stimulation of the face) suggest the presence of dysesthesia/paresthesia and mechanical allodynia in the ligated IoN territory.

Chronic Constriction Injury of the Rat's Infraorbital Nerve IoN-CCI to Study Trigeminal Neuropathic Pain

This manuscript describes a rodent model to study trigeminal neuropathic pain. The methods described include surgical procedures to perform a chronic constriction injury of the rat&#8217;s infraorbital nerve and the post-surgical behavioral tests to measure the changes in spontaneous and evoked behavior that are indicative of persistent pain and allodynia.

삼차 신경 병증 성 통증을 연구하기 위해 쥐의 눈꺼풀의 신경 (이온 CCI)의 만성 수축 부상

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

쥐의 복부 대동맥의 수축을 통해 심장 리모델링의 모델

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

Histological Quantification of Chronic Myocardial Infarct in Rats

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ischemic injury. Following the initial acute phase, chronic remodeling of the left ventricle (LV) modifies the structure and function of the heart. Permanent coronary ligation in small animals has been widely used as a reference model for a chronic model of MI. Thinning of the infarcted wall progressively develops to transmural fibrosis. Histological assessment of infarct size is commonly performed; nevertheless, a standardization of the methods for quantification is missing. Indeed, important methodological aspects, such as the number of sections analyzed and the sampling and quantification methods, are usually not described and therefore preclude comparison across investigations. Too often, quantification is performed on a single section obtained at the level of the papillary muscles. Because novel strategies aimed at reducing infarct expansion and remodeling are under investigation, there is an important need for the standardization of accurate heart sampling protocols. We describe an accurate method to quantify the infarct size using a systematic sampling of harvested rat heart and image analyses of trichromatic stained histological sections obtained from base to apex. We also provide evidence that calculating the expansion index (EI) allowed for infarct size assessment, taking into account changes of the left ventricle throughout the remodeling.

The post-mortem assessment of myocardial infarction (MI) in rodents is based on quantification of the infarct on stained heart sections. We describe an accurate method to quantify the infarct size using systematic sampling of harvested rat hearts from base to apex and image analyses of trichrome-stained histological sections.

쥐의 만성 심근 경색의 조직 학적 정량

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ...

Minimally Invasive Transverse Aortic Constriction in Mice

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard open-chest transverse aortic constriction (TAC). Although transverse aortic constriction is a highly functional method for the induction of high pressure in the left ventricle, it is a more difficult and lengthy procedure due to its use of artificial ventilation with tracheal intubation. TAC is oftentimes also less survivable, as the newer method, MTAC, neither requires the cutting of the ribs and intercostal muscles nor tracheal intubation with a ventilation setup. In MTAC, as opposed to a thoracotomy to access to the chest cavity, the aortic arch is reached through a midline incision in the anterior neck. The thyroid is pulled back to reveal the sternal notch. The sternum is subsequently cut down to the second rib level, and the aortic arch is reached simply by separating the connective tissues and thymus. From there, a suture can be wrapped around the arch and tied with a spacer, and then the sternal cut and skin can be closed. MTAC is a much faster and less invasive way to induce left ventricular hypertension and enables the possibility for high-throughput studies. The success of the constriction can be verified using high-frequency trans-thoracic echocardiography, particularly color Doppler and pulsed-wave Doppler, to determine the flow velocities of the aortic arch and left and right carotid arteries, the dimension of the blood vessels, and the left ventricular function and morphology. A successful constriction will also trigger significant histopathological changes, such as cardiac muscle cell hypertrophy with interstitial and perivascular fibrosis. Here, the procedure of MTAC is described, demonstrating how the resulting flow changes in the carotid arteries can be examined with echocardiography, gross morphology, and histopathological changes in the heart.

Minimally invasive transverse aortic constriction (MTAC) conserves the essentials of regular transverse aortic constriction (TAC) while eliminating the use of a ventilator with tracheal intubation. It proves to be a highly desirable method for high-throughput studies on left ventricular overload, particularly in translational studies.

마우스의 최소 침습 가로 대동맥 수축

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

Subcutaneous Trigeminal Nerve Field Stimulation for Refractory Facial Pain

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. Neuromodulation techniques for chronic facial pain include deep brain stimulation and motor cortex stimulation, which are complex to perform. Subcutaneous nerve field stimulation is certified for chronic back pain and is the least invasive form of neuromodulation. We applied this technique to treat chronic and neuropathic trigeminal pain as an individual therapy concept. First, trial stimulation is performed. Subcutaneous leads are placed in the painful trigeminal dermatome under local anesthesia. The leads are connected to an external neurostimulator that applies constant stimulation. Patients undergo a 12 day outpatient trial to assess the effect of the stimulation. Electrodes are removed after the trial. If the patient reports pain reduction of at least 50% in intensity and/or attack frequency, a reduction in medication or increase in quality of life, permanent implantation is scheduled. New electrodes are implanted under general anesthesia and are subcutaneously tunneled to an infraclavicular internal pulse generator. Patients are able to turn stimulation on and off and to increase or decrease the stimulation amplitude as needed.
This technique represents a minimal invasive alternative to other more invasive means of neuromodulation for trigeminal pain such as motor cortex stimulation or deep brain stimulation.

Treating chronic or neuropathic facial pain can be challenging when medical or standard treatment fails. Subcutaneous nerve field stimulation is the least invasive form of neuromodulation and is used for chronic back pain. We applied this technology to treat chronic and neuropathic trigeminal facial pain.

난치성 안면 통증을위한 피하 삼차 신경장 자극

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. ...

Development of Recombinant Proteins to Treat Chronic Pain

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates for treating debilitating pain conditions due to their capacity to regulate aberrant neuro-immune interactions. However, physiologically they work in a network of various cytokines, and therefore their therapeutic effect may not be optimal when used as stand-alone drugs. To overcome this limitation, we developed a fusion protein of the anti-inflammatory cytokines IL4 and IL10. Here, we describe the methods for production and quality control of IL4-10 recombinant fusion protein and we test the effectiveness of the IL4-10 fusion protein to resolve pain in a mouse model of persistent inflammatory pain.

In this paper we provide details for the methods of production and quality control of an IL4-10 recombinant fusion protein. We also show how to test the effectiveness of this protein to resolve pain in a mouse model of inflammatory pain.

만성 통증 치료에 재조합 단백질의 개발

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates ...