이 작품은 부여 계약 번호 246556 (유럽 프로젝트 RBUCE-UP)에서 유럽 연합의 일곱 번째 프레임 워크 프로그램 (FP7/2007-2013)에서 자금이 지원되고, HandiMedEx 프랑스의 공공 투자위원회에 의해 할당. 마르셀 Bonay는 Chancellerie 데 Universités 드 파리 (다리 Poix), 상테 Respiratoire EN 퐁 드 Dotation 공들인 및 센터 디부 지원 Respiratoire à 소재지 일드 프랑스 (CARDIF)에 의해 지원되었다

이 프로토콜은 우수 (파리 수드, 보조금 협정 제 246556 대학)와 Université 드 베르사유 생 캉탱 앙 이블린의 RBUCE-UP 의자의 윤리위원회의 승인을 받았다. 살균 수술 도구의 1. 준비 <ol><li> 실험실 세제로 수술 도구를 청소합니다. </li><li> 수술 전에 악기를 압력솥. </li><li> 수술 세션에서,이 수술 사이에 180 ° C에서 10 분 동안 뜨거운 구슬 살균기에 팁을 배치하여 도구를 소독. </li></ol> 약의 2. 준비 <ol><li> 사전 마취제 칵테일 및 수술 후 약물 x 1 2 ㎖ 주사기를 준비합니다. </li><li> Carprofen (5 ㎎ / ㎏), 부 프레 놀핀 (50 ㎍ / kg), Baytril (5 ㎎ / ㎏)과 Dexmedetomidine (0.5 ㎎ / ㎏) : 쥐의 무게에 따라, 사전 마취제와 주사기를 준비합니다. 락 테이트 화 링거 1 ML에 볼륨을 완료합니다. </li><li> 의 준비다른 주사기 사전 마취제의 반전 : Atipamezole (500 ㎍ / kg). </li></ol> 쥐 3. 마취 <ol><li> 동물에 2.2 단계에서 설명한 사전 마취 약물의 용액을 피하을 관리 할 수​​ 있습니다. 그런 다음, 새장에 다시 동물을 넣어 진정 효과가 나타날 때까지 기다립니다. </li><li> 100 % O 2 5 %의 이소 플루 란으로 채워 밀폐 된 챔버에 쥐를 놓고 호흡 리듬 (30 초 정도) 속도가 느려집니다 때까지 기다립니다. 그런 다음, 챔버에서 쥐를 제거하고 삽관 테이블에 배치합니다. </li></ol> 4. Orotracheal 삽관 <ol><li> 그의 뒤쪽에 동물을 거짓말, 다음 표에 자신의 앞니에 부착 된 스트랩을 배치하여 머리를 고정합니다. </li><li> 광섬유의 빛으로, 가슴 공간을 빛. 그런 다음, 동물의 입에 후두경 (또는 하나를 만들어 사용자 정의, 자세한 내용은 JOU 등. 20)를 배치합니다. 성대를 시각화. </li><li> 하락전자와 (성대 사이의) 기관에 orotracheal 가이드를 배치합니다. 가이드에 orotracheal 관 (16 G 카테터 크기)를 밀어 넣습니다. </li><li> 가이드를 제거하고 기관지 및하지 식도 관의 적절한 위치를 확인하고, 수분의 존재를 orotracheal 튜브의 끝에 배치 후두 미러와 검사. </li><li> (683 설치류 호흡기, 하버드 장치) 설치류 인공 호흡기에 튜브를 연결하고 (100 % O 2) 2 %에 이소 플루 란의 농도를 조정합니다. </li><li> 수술 테이프로 orotracheal 튜브를 고정합니다. </li></ol> 5. 척추 수술 <ol><li> 외과 의사 90 ° 각도에서 가리키는 코, 가열 수술 접시에 복부 욕창 위치에 동물을 배치합니다. 수술 전반에 걸쳐 37.5 ° C의 주위에 체온을 유지한다. </li><li> scapulas 사이 가위로 머리를 면도하고 거즈로 머리를 제거합니다. </li><li> 다음 7, 베타 딘으로 피부를 청소0 %의 알코올. 이 단계의 3 배를 반복합니다. </li><li> 발가락 핀치는 적절한 마취 깊이를 확인하기 위해 수술을 시작하기 전에 수행됩니다. &#160; 그런 다음, scapulas 사이 가위로 옆 피부 절개 rostro - 꼬리 쪽을 수행합니다. </li><li> 출혈을 방지하기 위해 건에 따라 acromiotrapezius 근육 rostro - 꼬리 쪽을 잘라. 다음 (척추 주변) spinalis 근육에 액세스 할 수 능형 근육을 떼어 놓다. </li><li> C3 척추에 C1에서 spinalis 근육을 철회. C2의 척추는 눈에 띄는 융기와 하나입니다. </li><li> 멸균 면봉을 사용하여 척추의 등 부분의 주위에 근육을 청소합니다. </li><li> rongeur으로 조심스럽게 C2의 융기를 제거하기 시작합니다. 등의 척수가 노출 될 때까지, 꼼꼼하게 계속합니다. 그 후궁 절제술는 등의 헤미의 후궁 절제술 있는지 확인하십시오. 이 지역의 주변에있는 척수 및 동맥을 둘러싸는 경질에주의를 기울이십시오. </li><li> # 55 포셉, ROS를 해부접근 금지 - 꼬리 쪽 C2 따라 두라는 다음 각 주동이와 꼬리쪽에 옆으로 계속합니다. </li><li> 뇌척수액을 스폰지. </li><li> microscissors와 자궁 경부 후근 2 번 아래 측면 부분을 확인합니다. 병변의 정도는 척수 (부상 등의보기에 대한 그림 2A 참조)의 중간 선에 도달 할만큼 가까이하는 마이크로 메스 확인합니다. 그렇지 않다면, 다른 컷 부상을 완료 할 수 있었다. 출혈의 경우, 멸균 면봉을 사용합니다. 그렇지 않으면, 동물이 부상에서 회복되지 않고 호흡을해야합니다, 반대쪽으로 이동하지 않도록주의하십시오. </li><li> 보호 층으로 근육을 봉합하고, 피부를 봉합 확대. 베타 딘 포화 멸균 거즈로 상처를 청소합니다. </li><li> 이소 플루 란 기화기을 끄고 반전 약물 주입 (Atipamezole을 [500 ㎍ / kg, IM), 체온을 확인합니다. </li><li> 동물은 인공 호흡기에 호흡을 시작하면,인공 호흡기에서 기관 튜브를 분리 한 다음 orotracheal 튜브를 제거합니다. 복구를위한 가열 새장에 동물을 배치합니다. </li></ol> 6. 수술 후 관리 수술 후, 동물은 복구를위한 최상의 환경을 보장하기 위해 지속적으로 모니터링됩니다. 항생제 (Baytril, 5 ㎎ / ㎏), 항 염증 (Carprofen, 5 ㎎ / ㎏)와 프레 놀핀은 (50 ㎍ / kg) 약은 감염을 방지하고 발생을 줄이기 위해 처음 2 일째에 수술마다 12 시간을 부여 수술 후 통증. 쥐 부드러운 음식과 물 (또는 1 차 수술 후 하루 jellified 물)에 대한 액세스 임의로있다. 피하 유체는 처음 몇 수술 일에 탈수를 방지하기 위해 사용될 수있다. 체중 및 음식 섭취량은 매일 모니터링된다. 이들의 환경은 실험 시간 후 상해 (자신의 새장 이중 주택, 튜브)에 걸쳐 충실.

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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motoneuron+BDNF/TrkB+signaling+enhances+functional+recovery+after+cervical+spinal+cord+injury.">Motoneuron BDNF/TrkB signaling enhances functional recovery after cervical spinal cord injury.</a> Exp Neurol. 247, 101-109 (2013).</li><li>Vinit, S., Darlot, F., Aoulaiche, H., Boulenguez, P., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+expression+of+c-Jun+and+HSP27+in+axotomized+and+spared+bulbospinal+neurons+after+cervical+spinal+cord+injury.">Distinct expression of c-Jun and HSP27 in axotomized and spared bulbospinal neurons after cervical spinal cord injury.</a> J Mol Neurosci. 45, 119-133 (2011).</li><li>Windelborn, J. A., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+activation+in+the+spinal+ventral+horn+caudal+to+cervical+injury.">Glial activation in the spinal ventral horn caudal to cervical injury.</a> Respir Physiol Neurobiol. 180, 61-68 (2012).</li><li>Vinit, S., Stamegna, J. C., Boulenguez, P., Gauthier, P., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restorative+respiratory+pathways+after+partial+cervical+spinal+cord+injury:+role+of+ipsilateral+phrenic+afferents.">Restorative respiratory pathways after partial cervical spinal cord injury: role of ipsilateral phrenic afferents.</a> Eur J Neurosci. 25, 3551-3560 (2007).</li><li>Dougherty, B. 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=Recovery+of+inspiratory+intercostal+muscle+activity+following+high+cervical+hemisection.">Recovery of inspiratory intercostal muscle activity following high cervical hemisection.</a> Respir Physiol Neurobiol. 183, 186-192 (2012).</li></ol>

저자는 더 경쟁 재정적 이익이 없다는 것을 선언합니다.

C2 상해 모델을 만들기의 기술적 인 문제 C2 손상 쥐 모델은 호흡 후 병변 신경 가소성을 연구하는 재미있는 도구입니다. 그러나, 재현성 및 신뢰성있는 모델을 생성하기 위해 필요한 단계는 다수이고 각 하나 연구의 결과에 영향을 미칠 수있다. 예를 들어, 삽관 과정에서 극도의주의가 orotracheal 튜브 등으로 인해 부상 자체 초기 호흡 부전 이외에 폐쇄성 호흡기 ?...

척수 외상은 영구적 인 마비와 같은 극적인 발생률과 인구에서 관찰 된 일반적인 부상입니다. 그러나 부상의 심각성은 초기 외상의 수준과 정도에 따라 달라집니다. 호흡 부전은 상부 경추 척수 손상 (SCI) 1 다음과 사망의 주요 원인이다. 현재 유일한 치료 치료는 인공 호흡기의 도움하에 환자를 배치하는 것입니다. 소수의 환자로 인해 포스트 병변 지연 발생 자발적 회복, 환기 지원 2를 투약 할 수 있기 때문에, 새로운 혁신적인 비 침습 치료를 개발할 필요가 긴급 3입니다. , 상상 속 치료 전략의 응용 프로그램을 공부하고, 따라서 호흡 부전에 자궁 SCI의 효과를 조사하는 것이 표준화 된 전 임상 모델을 갖는 것은 필수적이다. 이 기술 문서에서는, 우리는 특정 전임상 쥐 모델 (을)를 설명F 호흡 장애는 C2 수준에서 부분 자궁 SCI에 의해 유도. 이 모델은 현재 (: 4-13 후기) 세계의 여러 실험실에서 사용됩니다. 그러나, 수술 과정에서 약간의 차이가 특정 경추 부상 뮤린 모델을 생성하기 위해 다른 연구자간에 관찰 될 수있다. 호흡 출력에 C2 SCI의 효과는 첫 번째 포터 (14)에 의해 1895 년에 설명했다. 자궁 hemisection 침묵 횡격막 신경의 활동과 이후의 다이어프램 마비로 이어지는, 부상의 동측에 (뇌간, 그림 1A에 rVRG에 위치) 중앙 드라이브에서 횡격막 운동 신경원의 deafferentation을 유도한다. 반대쪽은 그대로 유지하고 동물이 생존 할 수 있습니다. 낮은 척추 세그먼트 (C4 레벨 15에서 예를 들어 contusive 부상)에있는 다른 SCI는 달리, 양측 횡격막 motoneuron 핵의 무결성이 유지됩니다. cerv 후iCal의 C2 부상, 일부 자발적인 활동으로 인해 분절 수준 C3-C6 (교차 횡격막 경로, CPP, 그림 1B)에서 척추 중앙선을 넘어 반대편 침묵 시냅스 경로의 활성화에 (횡격막과 다이어프램) 동측에 관찰 할 수있다 . 인 CPP의 활성화는, 정의에 의해, 동측 부분 횡격막 신경의 회복을 유도 반대측 phrenicotomy와 결합 된 C2의 hemisection은 시간에서 주 후 부상 16-18로 발생할 수 있습니다. 호흡 회복이 CPP 통로의 실제의 유익한 효과는 (19)를 한정하고 추가 조사와 치료는 자발적 회복 (3)의 크기를 향상시키기 위해 개발되어야한다. 이 프로토콜은 (이전과 횡격막 운동 신경원의 interneurons, 분자 및 cellula에서 호흡 생리학 다양한 수준에서 호흡 후 병변 가소성을 연구하는 전 임상 쥐 모델의 강력한 유형을 제공R, 예컨대 앞다리)의 운동뿐만 아니라 C2 부분 경추 척수 손상 후 호흡기 및 전위의 회복을 개선하는 목적 침습 및 비 침습성 치료 전략을 시험하는 모델.

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:572px;">Animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Male Sprague Dawley Rat</td>
			<td>Janvier</td>
			<td></td>
			<td>225-250g</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;">Surgical Instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Student Dumont #5 forceps</td>
			<td>Fine Science Tool</td>
			<td>91150-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Student Standard Pattern Forceps</td>
			<td>Fine Science Tool</td>
			<td>91100-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mayo-Stille Scissors</td>
			<td>Fine Science Tool</td>
			<td>14013-15</td>
			<td>Curved</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Student Vannas Spring Scissors</td>
			<td>Fine Science Tool</td>
			<td>91500-09</td>
			<td>Straight</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 8 mm Blades</td>
			<td>Fine Science Tool</td>
			<td>15025-10</td>
			<td>Straight Blunt/Blunt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Friedman Pearson Rongeur</td>
			<td>Fine Science Tool</td>
			<td>16121-14</td>
			<td>Curved</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissecting Knife - Fine Tip</td>
			<td>Fine Science Tool</td>
			<td>10055-12</td>
			<td>Straight</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Olsen-Hegar Needle Holder</td>
			<td>Fine Science Tool</td>
			<td>12002-14</td>
			<td>Serrated</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Weitlaner-Locktite Retractor</td>
			<td>Fine Science Tool</td>
			<td>17012-11</td>
			<td>2x3 Blunt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Absorbable surgical sutures</td>
			<td>Centravet</td>
			<td>BYO001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;">Equipment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hot Bead Steriliser</td>
			<td>Fine Science Tool</td>
			<td>18000-45</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Catheter&#160;</td>
			<td>Centravet</td>
			<td>CAT188</td>
			<td>16 gauge</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laryngoscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Guide wire</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laryngeal mirror</td>
			<td>Centravet</td>
			<td>MIR011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lactated Ringers</td>
			<td>Centravet</td>
			<td>RIN020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringe</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Needle</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">O2</td>
			<td>Air Liquid</td>
			<td>I1001M20R2A001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">683 RodentT Ventilator 115/230V</td>
			<td>Havard Apparatus</td>
			<td>55-0000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stand-Alone Vaporizer</td>
			<td>WPI</td>
			<td>EZ-155</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Thin line heated bed</td>
			<td>WPI</td>
			<td>EZ-211</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Air canister</td>
			<td>WPI</td>
			<td>EZ-258</td>
			<td></td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;">Drugs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Carprofen</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rimadyl</td>
			<td>Centravet</td>
			<td>RIM011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine</td>
			<td>Centravet</td>
			<td>BUP001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Baytril</td>
			<td>Centravet</td>
			<td>BAY001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dexmedetomidine</td>
			<td>Centravet</td>
			<td>DEX010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Atipamezole</td>
			<td>Centravet</td>
			<td>ANT201</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Betadine Solution</td>
			<td>Centravet</td>
			<td>VET002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Centravet</td>
			<td>VET066</td>
			<td></td>
		</tr>
	</tbody>
</table>

a murine model of cervical spinal cord injury to study post-lesional respiratory neuroplasticity

자궁 경부 척수 손상은 영구적 인 마비를 유도하고, 종종 호흡 곤란에 이르게한다. 지금까지, 더 효율적인 치료제가 높은 자궁 경부 척수 손상 (SCI) 다음 호흡 부전을 개선 / 향상을 위해 개발되지 않았습니다. 여기에서 우리는 다양한 사후 병변 호흡 신경 가소성을 연구하는 자궁 경부 2 (C2) metameric 수준 높은 SCI의 쥐의 전 임상 모델을 제안한다. 이 기술로 인해 뇌간에있는 호흡 센터에서 횡격막 운동 신경원의 deafferentation에 다이어프램의 hemiparalysis을 유도 할 것이다 C2 수준의 수술 부분 부상으로 구성되어 있습니다. 부상의 반대쪽은 그대로 유지하고 동물 복구를 할 수 있습니다. (흉추 및 요추 수준에서) 전위의 기능에 영향을 미치는 다른 SCIS는 달리, 호흡 기능 동물 동기 부여를 필요로하지 않고, 적자 / 복구의 정량화 쉽게 (다이어프램 횡격막 신경 녹음을 수행 할 수 있습니다의, 몸 전체 환기). 이 전 임상 C2 SCI 모델은 서로 다른 수준 (생리학 분자)에서 다양한 호흡 및 비 호흡 신경 가소성 이벤트를 공부하고있는 호흡을 향상시킬 다양한 상상 속 치료 전략을 테스트하기 위해 강력하고 유용하고 신뢰할 수있는 전임상 모델 SCI 환자.

부상의 정도 성공이 특정 실험 모델의 재현성 각 조작 / 외과 의사의 경험에 따라 달라집니다. 나머지 ventrolateral는 흰색 물질 21을 아끼지과 C2 부상 다음 호흡 복구 (횡격막 신경 활동과 다이어프램 활동)의 연속적인 양의 상관 관계가있다. 부상은 &quot;핸드 메이드&quot;이고 외과에서 연습을 필요로하기 때문에, 각 부상의 정도는 정확한 크기를 결정하기 ?...

Watch this Scientific Journal Video about A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity at JoVE.com

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

호흡 장애는 경추 척수 손상 후 사망의 주요 원인이다. 부분 자궁 경부 손상에 의해 유도 된 호흡의, 재생 가능한 정량화하고, 신뢰할 수있는 전임상 동물 모델을 갖는 것은 이후 호흡 및 비 호흡 신경 가소성을 이해하는 데 도움이 및 추정 복구 전략을 테스트 할 수 있습니다.

후 병변 호흡 신경 가소성을 연구하기 위해 자궁 경부 척수 손상의 생쥐 모델

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

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13.4K 조회수.  Université de Versailles Saint-Quentin-en-Yvelines.  호흡 장애는 경추 척수 손상 후 사망의 주요 원인이다. 부분 자궁 경부 손상에 의해 유도 된 호흡의, 재생 가능한 정량화하고, 신뢰할 수있는 전임상 동물 모델을 갖는 것은 이후 호흡 및 비 호흡 신경 가소성을 이해하는 데 도움이 및 추정 복구 전략을 테스트 할 수 있습니다. 

비디오: A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

루게릭병과 외상 척수 상해에서 자궁암 복부 호른를 타겟팅하는 Intraspinal 세포 이식

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

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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 ...

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

무한 호라이즌 임팩터을 사용하여 일방적 경부 척수의 부상의 Contusive 모델

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

마우스의 제어 자궁 경부 열상 상해

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

쥐에 심각한 척수 손상의 타박상 모델

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

만성 척수 부상으로 개인의 호흡기 모터 제어 평가 (RMCA)를 사용하여 호흡 근육 활성화의 평가

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

외상성 뇌 손상에 대한 제어 피질 충격 모델

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

무선 원격 측정 시스템은 척수 절개 및 배아 신경 세포 이식 줄기와 흰쥐의 심장 혈관 기능을 모니터링 할 수

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

실험 경추 척수 손상의 신경 전구체 세포 및 자기 조립 펩티드의 시너지 사용

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

척수 압축 부상의 보정 집게 모델

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...