우리는 원래 참조 11 페이지 130에 출연 microinjection의 피펫 제조 수치를 게시할 수있는 권한에 대한 Humana 보도 감사, videography에 대한 래리 Dlugas와 스티븐 왕, 교육 통신 OHSU학과, 영상 디자인 및 편집 래리 Dlugas; 아담 M. O &#39;퀸, 수석 디자이너, 기술적인 설계도를 제공하기 위해 우리의 정의 수평 층류 후드와 레 금세 공인을 디자인 Trion / Envirco, 빅터 Monterroso, 뮤직 비디오, 석사, 박사와 톰 Chatkupt, DVM, 비교 의학 OHSU학과,지도에 대한 우리 함께 동물 보호 프로토콜, 수술 기법 및 예방 진통제 처방, 마르셀 Perret-Gentil, DVM, MS, 수의학 suturing 기법에 그의 유인물을 공유하기위한, 우리의 어도비 프리미어 프로 영상 현미경 컴퓨터 워크 스테이션을 설계 에드워드 Porsov, 석사, 그리고 레아 공주와 LNS의 요나 Hinckley은 (포틀랜드 오레곤) 자막. 이 작품은 난청과 othe에있는 국립 연구소에서 교부금에 의해 지원되었다R 통신 장애 : DC R01 008595 및 DC R01 008595-04S2 (JB까지)와 P30 DC005983 (오레곤 청각 연구 센터 코어 그랜트, 피터 길레스피, 교장 탐정).

1. 복부 개복술 <ol><li> 나트륨 pentobarbital 마취 용액 (g 체중 당 7.5 μL)의 intraperitoneal 주사에 의한, 누구의 배아 배아 일 11.5 (질 플러그인이 감지 당일 정오가 배아 발달의 일 0.5입니다 E11.5)에있는 댐을 마취. 마취 용액 근무 :; 절대 에탄올 100 μL, 65 밀리그램 / ML 수성 마그네슘 황산 (자궁 음색을 modulates) 320 μL, 50 밀리그램 / ML pentobarbital 나트륨 용액 180 μL 및 프로필렌 글리콜 400 μL (와 차량 miscible을 수용액 및 유기 구성 요소). </li><li> 유해 자극 테스트를 실시하여 마취의 완전성을 평가 : 발자국 조르기, 꼬리 핀치, 뺨 및 vibrissae 터치 및 깜박임 반응. 각막에 무균 안과 연고를 바릅니다. </li><li> suprapubic 지역에서 가위로 흉곽과 훌륭한 블레이드 (Oster # 40 칼날)로 복부 모피 면도. 70 % 에탄올과 복부, 10 % povidone의 요오드 (Betadine)를 소독차 70 % 에탄올, 순차적으로. 장소는 무균 드레이프에서 마우스 복부 쪽 아래로 누른 다음 2-5 분을위한 가열 패드 또는 따뜻한 플레이트 (37 ° C)에서 설정합니다. </li><li> 공 - 스쳐 가위로 복부 중간선에서 복부 피부를 절개하다. 10~14mm에 대한 절개를 확장합니다. linea 알바, 복부 벽 배면의 중간선을 따라 avascular, 흰색 결합 조직 밴드를 식별합니다. 공이 가위 밀고로 linea 알바를 절개하고 절개 10~14밀리미터을 확장. 즉시 prewarmed (37 ° C) lactated 울리는의 솔루션으로 복부를 관개하다. </li><li> 주입 사이트에 과도한 압력을 적용하지 않고 고리 집게와 자궁의 두 뿔을를 외면 화하다. prewarmed, lactated 울리는의 솔루션 externalized 자궁 뿔이 관개. </li></ol> 2. Transuterine Microinjection <ol><li> 12-16 μ : 다음과 같은 특징을 가진 두꺼운 벽으로 둘러싸인, borosilicate 유리 모세관의 microinjection 피펫을 조작m 외경과 20 정도 베벨. ; 압력 = 200; 속도 = 46; = 0를 당길 시간 = 100) 열 = 램프 테스트 플러스 3 단위 : 작은 상자 필라멘트와 셔터 P-97 피펫 풀러에서 다음 프로그램을 사용합니다. 셔터 BV-10 beveler로 대구경 pipettes 위해 104C (금색) 연마 디스크를 사용합니다. 칼슘을 무료로 인산염의 3-4 μg / μL에서 플라스미드 Resuspend 표현이 (산도 7.2-7.4) 염분 버퍼. 30 초 동안 부드럽게 농축 DNA, 와동에 결정 신속한 녹색을 추가하고 10 초간 10,000g에 스핀. 시각 주사의 효능을 추적하는 데 필요한 빠른 그린의 최소 금액은 경험적으로 결정됩니다. DNA / 빠른 그린 솔루션 beveled 피펫을 Backfill. 압력 주입기 (Picospritzer 소스 가스 등&gt; 99% 순수한 질소를 사용하여)의 피펫 홀더에 로드된 microinjection 피펫에 연결합니다. </li><li> 낮은 강도, 그 주입 사이트 내에서 배아를 시각화하는 할로겐 라이트와 함께 자궁을 Transilluminate. 비 확인힘들 심장, 뇌 vesicles, 사지 꽃봉오리, hindbrain의 초기 4 번째 뇌실, 그리고 눈. prewarmed로 매 2 분, 수화를 유지하기 위해 울리는의 솔루션을 lactated 자궁을 관개하다. 배아의 방향과 해부 학적 경계표 위에서 언급 식별 자궁에 부드러운 압력을 적용합니다. </li><li> 동양은 배아 앞부분과 후부 가지의 측면 otocyst가있는 mesenchymal 지역 주요 헤드 정맥을 식별합니다. 적절한 otocyst은 자궁 transillumination로 볼 수 없습니다. otocyst은 정맥의 메인 트렁크와 함께 미국의 풋볼 경기에서 uprights 또는 goalposts의 모양을 형성, 주요 헤드 정맥의 앞부분과 후부 지점 사이의 중간에 위치하고 있습니다. otocyst는 uprights 사이의 중간입니다. </li><li> otocyst의 추정 위치와 라인의 궤도에있는 자궁을 통해 주입 피펫을 삽입합니다. uter 통과 후 한번 microinjector를 펄스우리는 추적 염료 및 피펫 팁의 대략적인 위치를 시각화합니다. 마이크로 미터 제어에서 피펫을 향상하고 깊이를 평가하기 위해 다시 펄스. 또한 반복 측면 헤드 mesenchyme 및 펄스로 피펫을 향상. 성공 otocyst 타겟팅 dorsally endolymphatic 덕트와 가리비 현관의 껍질 모양의 테이퍼 모양을 보여줄 것입니다. 자궁에 압력을 풀어 한 동작으로 배아 / 자궁에서 피펫을 제거하고 즉시 prewarmed와 자궁을 관개, 울리는의 솔루션 lactated. </li></ol> 3. 생체내의 Electroporation에 <ol><li> lactated 울리는의 솔루션과 함께 자궁을 관개하다. 갓 적용 lactated 울리는의 솔루션은 전기 부부 패들 스타일 전극에 자궁에 필요합니다. lactated 울리는의와 전극의 텅스텐 표면을 축축하게하다. 전극의 경로 센터 주입 otocyst합니다. 부드럽게 electroporation 파와 자궁을 압축ddles. 음극은 주입 otocyst과 양극이 uninjected otocyst에 인접한 자궁 벽과 접촉에 대한 측면 질 벽에와 접촉이다. electroporator에 발 페달 스위치와 함께 사각형 웨이브 펄스 열차를 실행. Electroporation 매개 변수는 다음과 같습니다 펄스 당 43 볼트와 950 밀리초 interpulse 지연에서 5, 50 밀리초 펄스. 펄스 트레인이 게재된 직후 자궁을 관개하다. 60-100 mAmps 귀의 상피 progenitors을 transfect 충분히 : 조직에 전달 전류가 기록합니다. 주사와 댐 당 4-6 E11.5의 배아를 electroporate. </li><li> 그의 내면의 귀있는 그 배아들 중 4 번째 뇌실로 수성 형광 dextran (알렉사 형석 488 플라스미드 표현식이 녹색 형광 단백질을 암호화하는 경우 플라스미드 표현식이 빨간색 형광 단백질이나 알렉스 형석 594을 암호화하는 경우)의 두 번째 독립 transuterine microinjection을 수행 정확하게 주입 및 펄스 당 전류의 최소 60 mAmps이 드했습니다electroporation시 livered. 형광 dextran은 hindbrain에서 출생에서 감지하게 내면의 귀 (3.5 참조) embryogenesis 동안 조작되었다 새끼의 선택을 가능하게합니다. </li><li> lactated 울리는의와 자궁 뿔이 관개. 복강으로 자궁 뿔을 꽂아. prewarmed, lactated 울리는의 솔루션 중 2-4 ML과 자궁 캐비티를 플러시하고 오버플로가 무균 드레이프로 절개 사이트 밖으로 배수 수 있습니다. 건조, 살균 소재 draping를 교체합니다. 비 절삭 바늘과 6-0 resorbable 봉합사와 복부 벽 봉합해. 우리는 복벽과 피부 모두에 대해, 다른 모든 바늘을 잠금, 실행 바늘을 선호합니다. </li><li> 댐 &#39;털&#39;을 건조하고 피하 주사에 의해 비 steroidal 안티 - 염증성 같은 Meloxicam으로 관리할 수 있습니다. 멸균 드레이프에 prewarmed 복구 케이지에 댐을 반환합니다. 모니터와 피묻은 방전을위한 댐 &#39;호흡, 절개 사이트 명백하고, 질을 기록합니다. 출혈은 라스입니다다시 있으나, 현재와 줄지 않는 경우에 그녀가 마취하에있는 동안, 댐을 안락사. 그녀는 의식과 ambulate하려는 시도을 차릴 시간을 확인합니다. 그녀가 먹고 마시의 흔적을 보여줍니다과 둥지가 만들기 시작되면 마우스 식민지로 댐을 반환합니다. 일반적으로 이것은 12 시간 내에 발생합니다. </li><li> 출생시 (출생 후의 일 0), 플래시 GFP 또는 텍사스 레드 필터로 stereofluorescence 해부 현미경을 사용하여 형광 dextran을 감지할 쓰레기의 각 강아지의 hindbrain 지역은 적절하게 설정합니다. 오직 lactating 댐 hindbrain 라벨을 표시 그 새끼로 돌아가기. </li></ol> 4. 대표 결과 <img src="/files/ftp_upload/3653/3653fig1.jpg" alt="그림 1" /> 1 그림. 개발 달팽이관에 Electroporation - 매개 유전자 전달. E11.5 otocyst은 (펄스 tra 표현 플라스미드 인코딩 향상된 녹색 형광 단백질 (EGFP)와 electroporated 맞았 었소매개 변수 : 다섯, 50 밀리초 / 펄스와 950 밀리초 interpulse 지연)에서 43 볼트 104. otocyst 달팽이관의 중간 회전을 통해 기지에서 EGFP 표정을 보여주 E11.5에 주입하고 electroporated되었다 강아지 출생 후의 일 6 (P6)에서) 담당자 내이 (内耳). 달팽이관의 측면 벽은 중간 턴하 꼭대기에서만 제거되었습니다. 꼭대기에 야기할 E11.5의 progenitors은 transfected되지 않았습니다. B)에 달팽이관의 immunostaining 전체 마운트는 () 헤어 셀 표식으로 마이 오신의 7A (Myo7a)는 해당 EGFP 표현은 Corti의 기관의 궤도를 따라하고 조잡한 머리카락 세포 베어링 감각 상피에 현지 나타냅니다. otocyst E11.5에 주입하고 electroporated되었다 E18.5 배아로부터 C) 대표 내이 (内耳). 레이저 공촛점 투영은 Myo7a - 긍정적인 감각 세포에서 EGFP 표현을 보여줍니다. EGFP expressio을 나타내는 Corti의 E18.5 기관에서 달팽이 감각 상피의 D) 레이저 공촛점 투영N 내부 세포의 (ihc), 바깥쪽 세포 (ohc), 내부 phalangial 세포 (IPC), 기둥 세포 (PC) 및 Deiters &#39;세포 (DC). (B)의 규모가 막대는 ()에 적용됩니다.

<ol>
	<li>Gillespie, P. G., Muller, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanotransduction+by+hair+cells:+models,+molecules,+and+mechanisms.">Mechanotransduction by hair cells: models, molecules, and mechanisms.</a> Cell. 139, 33-44 (2009).</li><li>Bok, J., Chang, W., Wu, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterning+and+morphogenesis+of+the+vertebrate+inner+ear.">Patterning and morphogenesis of the vertebrate inner ear.</a> Int. J. Dev. Biol. 51, 521-533 (2007).</li><li>Kelley, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+cell+fate+in+the+sensory+epithelia+of+the+inner+ear.">Regulation of cell fate in the sensory epithelia of the inner ear.</a> Nat. Rev. Neurosci. 7, 837-849 (2006).</li><li>Ohyama, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=BMP+signaling+is+necessary+for+patterning+the+sensory+and+nonsensory+regions+of+the+developing+mammalian+cochlea.">BMP signaling is necessary for patterning the sensory and nonsensory regions of the developing mammalian cochlea.</a> J. Neurosci. 30, 15044-15051 (2010).</li><li>Pan, W., Jin, Y., Stanger, B., Kiernan, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch+signaling+is+required+for+the+generation+of+hair+cells+and+supporting+cells+in+the+mammalian+inner+ear.">Notch signaling is required for the generation of hair cells and supporting cells in the mammalian inner ear.</a> Proc. Natl. Acad. Sci. U.S.A. 107, 15798-15803 (2010).</li><li>Gubbels, S. P., Woessner, D. W., Mitchell, J. C., Ricci, A. J., Brigande, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+auditory+hair+cells+produced+in+the+mammalian+cochlea+by+in+utero+gene+transfer.">Functional auditory hair cells produced in the mammalian cochlea by in utero gene transfer.</a> Nature. 455, 537-541 (2008).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the Care and Use of Laboratory Animals. , (2010).</li><li>Matsuda, T., Cepko, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+expression+of+transgenes+introduced+by+in+vivo+electroporation.">Controlled expression of transgenes introduced by in vivo electroporation.</a> Proc. Natl. Acad. Sci. U.S.A. 104, 1027-1032 (2007).</li><li>Chen, C., Smye, S. W., Robinson, M. P., Evans, 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=Membrane+electroporation+theories:+a+review.">Membrane electroporation theories: a review.</a> Med. Biol. Eng. Comput. 44, 5-14 (2006).</li><li>Saito, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+electroporation+in+the+embryonic+mouse+central+nervous+system.">In vivo electroporation in the embryonic mouse central nervous system.</a> Nat. Protoc. 1, 1552-1558 (2006).</li><li>Brigande, J. V., Gubbels, S. P., Woessner, D. W., Jungwirth, J. J., Bresee, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electroporation-mediated+gene+transfer+to+the+developing+mouse+inner+ear.">Electroporation-mediated gene transfer to the developing mouse inner ear.</a> Methods Mol. Biol. 493, 125-139 (2009).</li><li>Morsli, H., Choo, D., Ryan, A., Johnson, R., Wu, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+the+mouse+inner+ear+and+origin+of+its+sensory+organs.">Development of the mouse inner ear and origin of its sensory organs.</a> J. Neurosci. 18, 3327-3335 (1998).</li><li>Sher, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+embryonic+and+postnatal+development+of+the+inner+ear+of+the+mouse.">The embryonic and postnatal development of the inner ear of the mouse.</a> Acta. Otolaryngol. , 1-77 (1971).</li><li>Sheffield, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+vector+tropism+for+supporting+cells+in+the+developing+murine+cochlea.">Viral vector tropism for supporting cells in the developing murine cochlea.</a> Hear Res. 277, 28-36 (2011).</li><li>Bedrosian, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+delivery+of+recombinant+viruses+to+the+fetal+murine+cochlea:+transduction+characteristics+and+long-term+effects+on+auditory+function.">In vivo delivery of recombinant viruses to the fetal murine cochlea: transduction characteristics and long-term effects on auditory function.</a> Mol. Ther. 14, 328-335 (2006).</li><li>Reisinger, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probing+the+functional+equivalence+of+otoferlin+and+synaptotagmin+1+in+exocytosis.">Probing the functional equivalence of otoferlin and synaptotagmin 1 in exocytosis.</a> J. Neurosci. 31, 4886-4895 (2011).</li><li>Magnani, E., Bartling, L., Hake, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+Gateway+to+MultiSite+Gateway+in+one+recombination+event.">From Gateway to MultiSite Gateway in one recombination event.</a> BMC Mol. Biol. 7, 46 (2006).</li><li>Perret-Gentil, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Principles of Veterinary Suturing. , .</li><li>Oesterle, 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> P-1000 &#38; P-97 Pipette Cookbook. , (2011).</li></ol>

관심의 어떠한 충돌 선언 없습니다.

개발 마우스 내이 (内耳)에 유전자 이체 : 마우스 내이 (内耳)는 postimplantation 개발 12,13의 첫 주 동안 귀의 placode에서 개발하고 있습니다. 배아 일 9.5 (E9.5)으로 placode는 invaginated했으며 otocyst 2라는 액체가 채워진 소포로 morphed. 소포에 귀의 엽 성의 전구 물질이 성숙한 내이 내의 감각과 nonsensory 세포뿐만 아니라 vestibular과 청각 감각 상피 조직에 기계적으로 민감한 세포?...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 마이크로 살균 사례 </td><td> ROBOZ </td><td> RS-9900a </td><td> 8X8.5X1.25 인치 </td></tr><tr><td> 공 - 스쳐 가위 </td><td> 파인 과학 도구 </td><td> 14109-09 </td><td></td></tr><tr><td> 링 포셉 </td><td> 파인 과학 도구 </td><td> 11106-09 </td><td> 4.8mm ID/6mm OD </td></tr><tr><td> Adson 조직 집게 </td><td> 파인 과학 도구 </td><td> 11027-12 </td><td></td></tr><tr><td> 니들 드라이버 </td><td> 파인 과학 도구 </td><td> 12502-12 </td><td></td></tr><tr><td> 알레르기 주사기 트레이 </td><td> Becton Dickison </td><td> 305,536 </td><td></td></tr><tr><td> 봉합해​​ 6-0 </td><td> Syneture </td><td> GL-889 </td><td> 0.7 메트릭 위장 봉합사 </td></tr><tr><td> Lactated 울리는의 사출 USP </td><td> 벡스터 </td><td> 2B2323 </td><td></td></tr><tr><td> 빠른 그린 </td><td> 시그마 알드리치 </td><td> F7258 </td><td></td></tr><tr><td> Borosilicate 유리 모세관 </td><td> 하버드 장치 </td><td> 30-0053 </td><td></td></tr><tr><td> Nembutal 나트륨 솔루션 </td><td> 박수 제약 주식 회사 </td><td> NDC 67386-501-52 </td><td></td></tr><tr><td> MgSO4.7H2O </td><td> 피셔 과학 </td><td> M63-500 </td><td></td></tr><tr><td> 프로필렌 글리콜 </td><td> 피셔 과학 </td><td> P355-1 </td><td></td></tr><tr><td> 에탄올 </td><td> 시그마 알드리치 </td><td> E7023-500 </td><td></td></tr><tr><td> Meloxicam </td><td> Boehringer Ingeheim </td><td> 나다 141-219 </td><td></td></tr><tr><td> Micropipette 풀러 </td><td> 셔터 인 스트 루먼트 </td><td> P-97 </td><td> FB255B 상자 필라멘트, 셔터 악기에서 피펫 요리책을 참고 </td></tr><tr><td> Microelectrode의 Beveler </td><td> 셔터 인 스트 루먼트 </td><td> BV-10 </td><td> 대형 pipettes를위한 104C beveling 디스크는, 이론을 beveling에 대한 설명서를 참조 </td></tr><tr><td> Micropipette 홀더 </td><td> 워너 인 스트 루먼트 </td><td> MP-S15T </td><td> Picospritzer 튜빙을위한 1.5mm 외경의 피펫과 여성의 압력 포트. </td></tr><tr><td> 족집게 스타일 전극 </td><td> Protech 인터내셔널 </td><td> CUY650P5 </td><td> 5mm 외경 </td></tr><tr><td> 스퀘어 웨이브 Electroporator </td><td> Protech 인터내셔널 </td><td> CUY21EDIT </td><td> Footpedal 권장 </td></tr><tr><td> PICOSPRITZER III </td><td> 파커 Hannifin </td><td> 051-0500-900 </td><td> Footpedal 권장 </td></tr><tr><td> 수동 제어 Micromanipulator </td><td> 하버드 장치 </td><td> 640,056 </td><td></td></tr><tr><td> 수평 층류 클린 벤치 </td><td> Envirco </td><td></td><td> LF 630-10554로 정의 수정. 후드의 약도에 대한 보충 정보를 참조하십시오. </td></tr><tr><td> Leica stereofluorescence는 Lumencor 솔라 라이트 엔진 microcope을 해부 </td><td> Bartels와 스타우트와 Lumencor </td><td> Lumencor 솔라 라이트 엔진과 MZ10F </td><td> MZ10F을 집중하고 솔라 라이트 엔진을 실행하는 데 Footpedals 추천합니다 </td></tr><tr><td> 알렉사 형석 594 Dextran </td><td> Invitrogen </td><td> D22913 </td><td> 10mg/ml, 수성 </td></tr><tr><td> 알렉사 형석 488 Dextran </td><td> Invitrogen </td><td> D22910 </td><td> 10mg/ml, 수성 </td></tr><tr><td> Enviro-dri </td><td> 목자 특수 아빠ERS </td><td></td><td> <a href="http://www.ssponline.com" target="_blank">www.ssponline.com</a> </td></tr></tbody></table>

gene transfer to the developing mouse inner ear by in vivo electroporation

; utricle 및 saccule의 maculae 및 코일 달팽이관에 Corti의 기관 반원의 운하 ampullae 3 cristae : 포유류의 내이 (内耳)는 6 독특한 감각 상피 조직이 있습니다. cristae와 maculae은 Corti의 기관에서 청각 세포가 1 청각의 기본 트랜스 듀서 반면 기계적 자극이 균형의 특별한 감각을 보조하다 할 transduce vestibular 세포가 포함되어 있습니다. 반원의 운하와 달팽이관 이러한 감각 상피 조직과 morphogenesis의 세포 운명 사양은 마우스의 태동의 두 번째 주 동안 자리를 대신하고 대부분이 출산 2,3 전에 완료됩니다. 마우스 내이 (内耳)의 발달 연구는 정기적으로 세포 및 / 또는 형태학의 phenotypes 4,5의 분자 기초에 대한 통찰력을 얻기 위해 다른 배아 또는 출생 후의 단계에서 유전자 변형 배아를 수확하여 실시하고 있습니다. 우리는 utero의 발전 마우스 내이 (内耳)에 해당 유전자 전달 가설 &lt;/ em&gt;는 이득과 손실의 기능 연구의 맥락에서 포유류의 내이 (内耳)의 개발 6 밑에있는 유전자 메커니즘의 심문에 대한 전통적인 마우스 transgenesis에 대한 무료 접근 방식을 나타냅니다. 복부 개복술 1), 2) transuterine microinjection 및 생체내의 electroporation 3) : 개발 마우스 내이 (内耳)에 유전자 misexpression 연구를 실시하는 실험적 패러다임 여기서 보여주 세 일반적인 단계로 해결됩니다. 복부 개복술은 이식 배아 7 실험적 액세스할 자궁 externalization를 허용 마우스 생존 수술 기법입니다. Transuterine microinjection은 귀의 소포 또는 otocyst의 루멘에 플라스미드 표현을 소개 beveled, 유리 모세관 micropipettes의 사용이다. 생체내에 electroporation은 전구 세포의 8-10로 플라스미드 표현을 운전 사각 파, 직류 펄스의 응용 프로그램입니다. 11의 각 단계에 대한 상세한 기록이 포함되어 있습니다. 마우스 실험 embryological 기술은 산문에서 배우고 스틸 이미지 혼자하기 어려울 수 있습니다. 현재 직장에서 우리는 유전자 전달 절차의 3 단계를 보여줍니다. 대부분 비판적으로, 우리는 방법 정확하게 표시하는 디지털 비디오 현미경을 배포 : 1) utero에서 배아 방향을 파악, 2) otocyst에 주사를 타겟팅을위한 배아를 방향, 3) 배아 일 11.5에서 otocyst로 추적 염료 용액과 혼합 DNA를 microinject과 12.5, 출생에서 출생 후의 선택과 5) 라벨 electroporated 배아, 4) 주입 otocyst을 electroporate. 일반적인 방법론적인 오류를 방지하는 방법을 논의; otocyst mistargeting의 가장 일반적인 원인에 그림 가이드, 우리는 성공적으로 transfected 안쪽 귀의 대표적인 예제를 제공하고 utero g에 쓰기 위해 현재 지침에너지 전송 동물 케어 프로토콜.

Watch this Scientific Journal Video about Gene Transfer to the Developing Mouse Inner Ear by In Vivo Electroporation at JoVE.com

Gene Transfer to the Developing Mouse Inner Ear by In Vivo Electroporation

마우스 내이 (内耳)는 누구의 발달 프로그램의 회임 기간 동안 정교하고 있습니다 placode 파생 감각 기관이다. 우리를 정의<em&gt; utero에서</em세 단계로 구성된&gt; 유전자 전달 기술 : 마우스 복부 개복술, transuterine microinjection, 그리고<em&gt; 생체내에서</em&gt; electroporation. 우리는 중요한 실험 embryological 기술을 입증하는 디지털 비디오 현미경을 사용합니다.

에 의해 개발 마우스 내이 (内耳)에 진 전송<em&gt; VIVO에서</em&gt; Electroporation

The mammalian inner ear has 6 distinct sensory epithelia: 3 cristae in the ampullae of the semicircular canals; maculae in the utricle and saccule; and ...

gene-transfer-to-developing-mouse-inner-ear-vivo

Research

JoVE Journal

Neuroscience

Gene Transfer to the Developing Mouse Inner Ear by In Vivo Electroporation

13.9K 조회수.  Oregon Health & Science University. 마우스 내이 (内耳)는 누구의 발달 프로그램의 회임 기간 동안 정교하고 있습니다 placode 파생 감각 기관이다. 우리를 정의 utero에서 유전자 전달 기술 : 마우스 복부 개복술, transuterine microinjection, 그리고 생체내에서 electroporation. 우리는 중요한 실험 embryological 기술을 입증하는 디지털 비디오 현미경을 사용합니다.

비디오: Gene Transfer to the Developing Mouse Inner Ear by In Vivo Electroporation

Gene Delivery to Postnatal Rat Brain by Non-ventricular Plasmid Injection and Electroporation

Creation of transgenic animals is a standard approach in studying functions of a gene of interest in vivo. However, many knockout or transgenic animals are not viable in those cases where the modified gene is expressed or deleted in the whole organism. Moreover, a variety of compensatory mechanisms often make it difficult to interpret the results. The compensatory effects can be alleviated by either timing the gene expression or limiting the amount of transfected cells.
The method of postnatal non-ventricular microinjection and in vivo electroporation allows targeted delivery of genes, siRNA or dye molecules directly to a small region of interest in the newborn rodent brain. In contrast to conventional ventricular injection technique, this method allows transfection of non-migratory cell types. Animals transfected by means of the method described here can be used, for example, for two-photon in vivo imaging or in electrophysiological experiments on acute brain slices.

This protocol describes a non-viral method of delivery of genetic constructs to a certain area of living rodent brain. The method consists of plasmid preparation, micropipette fabrication, neonatal rat pup surgery, microinjection of the construct, and in vivo electroporation.

비 심실 플라스미드 주입 및 Electroporation에 의해 출생 후의 쥐 뇌에 유전자 전달

Creation of transgenic animals is a standard approach in studying functions of a gene of interest in vivo. However, many knockout or transgenic animals ...

연구

신경과학

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

The targeting and refinement of RGC projections to the midbrain is a popular and powerful model system for studying how precise patterns of neural connectivity form during development. In mice, retinofugal projections are arranged in a topographic manner and form eye-specific layers in the Lateral Geniculate Nucleus (dLGN) of the thalamus and the Superior Colliculus (SC). The development of these precise patterns of retinofugal projections has typically been studied by labeling populations of RGCs with fluorescent dyes and tracers, such as horseradish peroxidase1-4. However, these methods are too coarse to provide insight into developmental changes in individual RGC axonal arbor morphology that are the basis of retinotopic map formation. They also do not allow for the genetic manipulation of RGCs.
Recently, electroporation has become an effective method for providing precise spatial and temporal control for delivery of charged molecules into the retina5-11. Current retinal electroporation protocols do not allow for genetic manipulation and tracing of retinofugal projections of a single or small cluster of RGCs in postnatal mice. It has been argued that postnatal in vivo electroporation is not a viable method for transfecting RGCs since the labeling efficiency is extremely low and hence requires targeting at embryonic ages when RGC progenitors are undergoing differentiation and proliferation6. 
In this video we describe an in vivo electroporation protocol for targeted delivery of genes, shRNA, and fluorescent dextrans to murine RGCs postnatally. This technique provides a cost effective, fast and relatively easy platform for efficient screening of candidate genes involved in several aspects of neural development including axon retraction, branching, lamination, regeneration and synapse formation at various stages of circuit development. In summary we describe here a valuable tool which will provide further insights into the molecular mechanisms underlying sensory map development.

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

We demonstrate an in vivo electroporation protocol for transfecting single or small clusters of retinal ganglion cells (RGCs) and other retinal cell types in postnatal mice over a wide range of ages. The ability to label and genetically manipulate postnatal RGCs in vivo is a powerful tool for developmental studies.

마우스 망막 신경절 세포 Transfection로 생체내에 Electroporation

The targeting and refinement of RGC projections to the midbrain is a popular and powerful model system for studying how precise patterns of neural ...

Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation

Transcription factors within cellular gene networks control the spatiotemporal pattern and levels of expression of their target genes by binding to cis-regulatory elements (CREs), short (&tilde;300-600 bp) stretches of genomic DNA which can lie upstream, downstream, or within the introns of the genes they control. CREs (i.e., enhancers/promoters) typically consist of multiple clustered binding sites for both transcriptional activators and repressors1-3. They serve as logical integrators of transcriptional input giving a unitary output in the form of spatiotemporally precise and quantitatively exact promoter activity. Most studies of mammalian cis-regulation to date have relied on mouse transgenesis as a means of assaying the enhancer function of CREs4-5. This technique is time-consuming, costly and, on account of insertion site effects, largely non-quantitative. On the other hand, quantitative assays for mammalian CRE function have been developed in tissue culture systems (e.g., dual luciferase assays), but the in vivo relevance of these results is often uncertain. 
Electroporation offers an excellent alternative to traditional mouse transgenesis in that it permits both spatiotemporal and quantitative assessment of cis-regulatory activity in living mammalian tissue. This technique has been particularly useful in the analysis of cis-regulation in the central nervous system, especially in the cerebral cortex and the retina6-8. While mouse retinal electroporation, both in vivo and ex vivo, has been developed and extensively described by Matsuda and Cepko6-7,9, we have recently developed a simple approach to quantify the activity of photoreceptor-specific CREs in electroporated mouse retinas10. Given that the amount of DNA that is introduced into the retina by electroporation can vary from experiment to experiment, it is necessary to include a co-electroporated 'loading control' in all experiments. In this respect, the technique is very similar to the dual luciferase assay used to quantify promoter activity in cultured cells. 
When assaying photoreceptor cis-regulatory activity, electroporation is usually performed in newborn mice (postnatal day 0, P0) which is the time of peak rod production11-12. Once retinal cell types become post-mitotic, electroporation is much less efficient. Given the high rate of rod birth in newborn mice and the fact that rods constitute more than 70% of the cells in the adult mouse retina, the majority of cells that are electroporated at P0 are rods. For this reason, rod photoreceptors are the easiest retinal cell type to study via electroporation. The technique we describe here is primarily useful for quantifying the activity of photoreceptor CREs.

Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation

This protocol describes a simple and inexpensive way to quantify the activity of cis-regulatory elements (i.e., enhancer/promoters) in living mouse retinas via explant electroporation. DNA preparation, retinal dissection, electroporation, retinal explant culture, and post-fixation analysis and quantification are described.

의 활동을 Quantifying CIS</emExplant Electroporation하여 마우스 망막에서> - 규제 요소

Transcription factors within cellular gene networks control the spatiotemporal pattern and levels of expression of their target genes by binding to ...

In vivo Electroporation of Developing Mouse Retina

The functional characterization of genes expressed during mammalian retinal development remains a significant challenge. Gene targeting to generate constitutive or conditional loss of function knockouts remains cost and labor intensive, as well as time consuming. Adding to these challenges, retina expressed genes may have essential roles outside the retina leading to unintended confounds when using a knockout approach. Furthermore, the ability to ectopically express a gene in a gain of function experiment can be extremely valuable when attempting to identify a role in cell fate specification and/or terminal differentiation.
We present a method for the rapid and efficient incorporation of DNA plasmids into the neonatal mouse retina by electroporation. The application of short electrical impulses above a certain field strength results in a transient increase in plasma membrane permeability, facilitating the transfer of material across the membrane 1,2,3,4. Groundbreaking work demonstrated that electroporation could be utilized as a method of gene transfer into mammalian cells by inducing the formation of hydrophilic plasma membrane pores allowing the passage of highly charged DNA through the lipid bilayer 5. Continuous technical development has resulted in the viability of electroporation as a method for in vivo gene transfer in multiple mouse tissues including the retina, the method for which is described herein 6, 7, 8, 9, 10. 
DNA solution is injected into the subretinal space so that DNA is placed between the retinal pigmented epithelium and retina of the neonatal (P0) mouse and electrical pulses are applied using a tweezer electrode. The lateral placement of the eyes in the mouse allows for the easy orientation of the tweezer electrode to the necessary negative pole-DNA-retina-positive pole alignment. Extensive incorporation and expression of transferred genes can be identified by postnatal day 2 (P2). Due to the lack of significant lateral migration of cells in the retina, electroporated and non-electroporated regions are generated. Non-electroporated regions may serve as internal histological controls where appropriate. 
Retinal electroporation can be used to express a gene under a ubiquitous promoter, such as CAG, or to disrupt gene function using shRNA constructs or Cre-recombinase. More targeted expression can be achieved by designing constructs with cell specific gene promoters. Visualization of electroporated cells is achieved using bicistronic constructs expressing GFP or by co-electroporating a GFP expression construct. Furthermore, multiple constructs may be electroporated for the study of combinatorial gene effects or simultaneous gain and loss of function of different genes. Retinal electroporation may also be utilized for the analysis of genomic cis-regulatory elements by generating appropriate expression constructs and deletion mutants. Such experiments can be used to identify cis-regulatory regions sufficient or required for cell specific gene expression 11. Potential experiments are limited only by construct availability.

In vivo Electroporation of Developing Mouse Retina

A method for the incorporation of plasmid DNA into murine retinal cells for the purpose of performing either gain- or loss of function studies in vivo is presented. This method capitalizes on the transient increase in permeability of cell plasma membranes induced by the application of an external electrical field.

개발 마우스 망막의 생체내 Electroporation에

The functional characterization of genes expressed during mammalian retinal development remains a significant challenge.  Gene targeting to generate ...

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

In order to understand the function of genes expressed in specific region of the developing brain, including signaling molecules and axon guidance molecules, local gene transfer or knock- out is required. Gene targeting knock-in or knock-out into local regions is possible to perform with combination with a specific CRE line, which is laborious, costly, and time consuming. Therefore, a simple transfection method, an in utero electroporation technique, which can be performed with short time, will be handy to test the possible function of candidate genes prior to the generation of transgenic animals 1,2. In addition to this, in utero electroporation targets areas of the brain where no specific CRE line exists, and will limit embryonic lethality 3,4. Here, we present a method of in utero electroporation combining two different types of electrodes for simple and convenient gene transfer into target areas of the developing brain. First, a unique holding method of embryos using an optic fiber optic light cable will make small embryos (from E9.5) visible for targeted DNA solution injection into ventricles and needle type electrodes insertion to the targeted brain area 5,6. The patterning of the brain such as cortical area occur at early embryonic stage, therefore, these early electroporation from E9.5 make a big contribution to understand entire area patterning event. Second, the precise shape of a capillary prevents uterine damage by making holes by insertion of the capillary. Furthermore, the precise shape of the needle electrodes are created with tungsten and platinum wire and sharpened using sand paper and insulated with nail polish 7, a method which is described in great detail in this protocol. This unique technique allows transfection of plasmid DNA into restricted areas of the brain and will enable small embryos to be electroporated. This will help to, open a new window for many scientists who are working on cell differentiation, cell migration, axon guidance in very early embryonic stage. Moreover, this technique will allow scientists to transfect plasmid DNA into deep parts of the developing brain such as thalamus and hypothalamus, where not many region-specific CRE lines exist for gain of function (GOF) or loss of function (LOF) analyses.

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

A gene transfer method into the developing mouse brain is described by using a unique surgical method and special shape of electrodes. This unique technique allows transfection of plasmid DNA temporally and spatially, which will aid many neuroscientists in studying brain development.

마우스 Utero에 Electroporation : 통제 Spatiotemporal 진 Transfection

In order to understand the function of genes expressed in specific region of the developing brain, including signaling molecules and axon guidance ...

Isolating LacZ-expressing Cells from Mouse Inner Ear Tissues using Flow Cytometry

Isolation of specific cell types allows one to analyze rare cell populations such as stem/progenitor cells. Such an approach to studying inner ear tissues presents a unique challenge because of the paucity of cells of interest and few transgenic reporter mouse models. Here, we describe a protocol using fluorescence-conjugated probes to selectively label LacZ-positive cells from the neonatal cochleae.
The most common underlying pathology of sensorineural hearing loss is the irreversible damage and loss of cochlear sensory hair cells, which are required to transduce sound waves to neural impulses. Recent evidence suggests that the murine auditory and vestibular organs harbor stem/progenitor cells that may have regenerative potential1,2. These findings warrant further investigation, including identifying specific cell types with stem/progenitor cell characteristics. The Wnt signaling pathway has been demonstrated to play a critical role in maintaining stem/progenitor cell populations in several organ systems3-7. We have recently identified Wnt-responsive Axin2-expressing cells in the neonatal cochlea, but their function is largely unknown8.
To better understand the behavior of these Wnt-responsive cells in vitro, we have developed a method of isolating Axin2-expressing cells from cochleae of Axin2-LacZ reporter mice9. Using flow cytometry to isolate Axin2-LacZ positive cells from the neonatal cochleae, we could in turn execute a variety of experiments on live cells to interrogate their behavior as stem/progenitor cells. Here, we describe in detail the steps for the microdissection of neonatal cochlea, dissociation of these tissues, labeling of the LacZ-positive cells using a fluorogenic substrate, and cell sorting. Techniques for dissociating cochleae into single cells and isolating cochlear cells via flow cytometry have been described2,10-12. We have made modifications to these techniques to establish a novel protocol to isolate LacZ-expressing cells from the neonatal cochlea.

Flow cytometry is a powerful tool allowing for the isolation and study of specific cell populations. This protocol describes steps for isolating LacZ-expressing cells from cochlear tissues from neonatal transgenic mice. Dissociated cochlear cells were labeled using fluorescent-conjugated substrates of &#946;-galactosidase prior to separation via flow cytometry.

분리 LacZ를 표현 유동세포계측법를 사용하여 마우스 내이 (内耳) 조직의 세포를

Isolation of specific cell types allows one to analyze rare cell populations such as stem/progenitor cells. Such an approach to studying inner ear tissues ...

High-resolution Live Imaging of Cell Behavior in the Developing Neuroepithelium

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central nervous system (CNS). Although much is known about the molecular mechanisms that pattern the spinal cord and elicit neuronal differentiation1, 2, we lack a deep understanding of these early events at the level of cell behavior. It is thus critical to study the behavior of neural progenitors in real time as they undergo neurogenesis.
In the past, real-time imaging of early embryonic tissue has been limited by cell/tissue viability in culture as well as the phototoxic effects of fluorescent imaging. Here we present a novel assay for imaging such tissue for long periods of time, utilizing a novel ex vivo slice culture protocol and wide-field fluorescence microscopy (Fig. 1). This approach achieves long-term time-lapse monitoring of chick embryonic spinal cord progenitor cells with high spatial and temporal resolution.
This assay may be modified to image a range of embryonic tissues3, 4 In addition to the observation of cellular and sub-cellular behaviors, the development of novel and highly sensitive reporters for gene activity (for example, Notch signaling5) makes this assay a powerful tool with which to understand how signaling regulates cell behavior during embryonic development.

Imaging embryonic tissue in real-time is challenging over long periods of time. Here we present an assay for monitoring cellular and sub-cellular changes in chick spinal cord for long periods with high spatial and temporal resolution. This technique can be adapted for other regions of the nervous system and developing embryo.

개발 Neuroepithelium의 세포 문제의 고해상도 라이브 영상

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central ...

Patch Clamp Recordings in Inner Ear Hair Cells Isolated from Zebrafish

Patch clamp analyses of the voltage-gated channels in sensory hair cells isolated from a variety of species have been described previously1-4 but this video represents the first application of those techniques to hair cells from zebrafish. Here we demonstrate a method to isolate healthy, intact hair cells from all of the inner ear end-organs: saccule, lagena, utricle and semicircular canals. Further, we demonstrate the diversity in hair cell size and morphology and give an example of the kinds of patch clamp recordings that can be obtained. The advantage of the use of this zebrafish model system over others stems from the availability of zebrafish mutants that affect both hearing and balance. In combination with the use of transgenic lines and other techniques that utilize genetic analysis and manipulation, the cell isolation and electrophysiological methods introduced here should facilitate greater insight into the roles hair cells play in mediating these sensory modalities.

The purpose of this video is to demonstrate procedures for obtaining healthy, intact hair cells from the inner ear organs of adult zebrafish and then using them for patch clamp studies aimed at characterizing the biophysical properties of their voltage-gated channels.

귀에 세포의 패치 클램프 녹음 절연 Zebrafish에서

Patch  clamp analyses of the voltage-gated channels in sensory hair cells isolated  from a variety of species have been described previously1-4 but this  ...

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

Genetic modification of specific regions of the developing mammalian brain is a very powerful experimental approach. However, generating novel mouse mutants is often frustratingly slow. It has been shown that access to the mouse brain developing in utero with reasonable post-operatory survival is possible. Still, results with this procedure have been reported almost exclusively for the most superficial and easily accessible part of the developing brain, i.e. the cortex. The thalamus, a narrower and more medial region, has proven more difficult to target. Transfection into deeper nuclei, especially those of the hypothalamus, is perhaps the most challenging and therefore very few results have been reported. Here we demonstrate a procedure to target the entire hypothalamic neuroepithelium or part of it (hypothalamic regions) for transfection through electroporation. The keys to our approach are longer narcosis times, injection in the third ventricle, and appropriate kind and positioning of the electrodes. Additionally, we show results of targeting and subsequent histological analysis of the most recessed hypothalamic nucleus, the mammillary body.

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

Despite the functional and medical importance of the hypothalamus, in utero genetic manipulation of its development has rarely been attempted. We show a detailed procedure for in utero electroporation into the mouse hypothalamus and show representative results of total and partial (regional) hypothalamic transfection.

마우스의 유전자 조작을 통해 시상 하부 개발<em&gt; 자궁 내</em&gt; 일렉트로

Genetic  modification of specific regions of the developing mammalian brain is a very  powerful experimental approach. However, generating novel mouse ...

Surgical Method for Virally Mediated Gene Delivery to the Mouse Inner Ear through the Round Window Membrane

Gene therapy, used to achieve functional recovery from sensorineural deafness, promises to grant better understanding of the underlying molecular and genetic mechanisms that contribute to hearing loss. Introduction of vectors into the inner ear must be done in a way that widely distributes the agent throughout the cochlea while minimizing injury to the existing structures. This manuscript describes a post-auricular surgical approach that can be used for mouse cochlear therapy using molecular, pharmacologic, and viral delivery to mice postnatal day 10 and older via the round window membrane (RWM). This surgical approach enables rapid and direct delivery into the scala tympani while minimizing blood loss and avoiding animal mortality. This technique involves negligible or no damage to essential structures of the inner and middle ear as well as neck muscles while wholly preserving hearing. To demonstrate the efficacy of this surgical technique, the vesicular glutamate transporter 3 knockout (VGLUT3 KO) mice will be used as an example of a mouse model of congenital deafness that recovers hearing after delivery of VGLUT3 to the inner ear using an adeno-associated virus (AAV-1).

The described post-auricular surgical approach allows rapid and direct delivery into the mouse cochlear scala tympani while minimizing blood loss and animal mortality. This method can be used for cochlear therapy using molecular, pharmacologic and viral delivery to postnatal mice through the round window membrane.

라운드 창 멤브레인을 통해 마우스 내부 귀에 바이러스로 매개 유전자 전달을위한 수술 방법

Gene therapy, used to achieve functional recovery from sensorineural deafness, promises to grant better understanding of the underlying molecular and ...