Name: intravenous injections in neonatal mice
Uploaded: 2014-11-11T13:00:00.000+00:00
Duration: 5 min 17 s
Description: Watch this Scientific Journal Video about Intravenous Injections in Neonatal Mice at JoVE.com

저자는 금융 지원을위한 SMA의 NINDS, FightSMA, 가족을 인정하고 싶습니다. SEGL은 NINDS 훈련 보조금 # 5T32NS077984-02에 의해 지원됩니다.

프로토콜에 나열된 모든 절차는 오하이오 주립 대학의 동물 사용 및 관리위원회 (IACUC) 연구소 승인되었습니다. 작업 1. 준비 <ol><li> 깔짚, 해부 현미경, 90 °로 주사 (광원의 사용 각도로 위치 될 수 광원으로부터 댐을 분리하는, 빈 케이지 마우스 새끼 마취 젖은 얼음 수집 &#160; 주사 부위에 각도는 정맥), 주사에 대한 동물을 배치 깨끗한 표면, 면봉, 3/8 &quot;30 G 바늘 (동물 당), 1 % 에반스 블루 염료 3/10 CC 인슐린 주사기 (가린다 교육을위한 인산염 완충 생리 식염수 (PBS)) 솔루션을했다. </li><li> 새끼를 조작하면서 별도의 케이지에 댐을 제거합니다. </li></ol> 2. 사출 절차 <ol><li> 동물을 마취하는 30 ~ 60 초 동안 얼음물에 직접 하나의 강아지를 놓습니다. t 방치하지 마십시오인해 심실 세동, 조직의 저산소증과 대사성 산증을 포함하여 저체온증과 관련된 합병증의 위험에 너무 오래 얼음에 그는 동물. 참고 : 우리의 경험은 30 ~ 60 초 분사를 허용하는 강아지의 움직임을 느리게하기에 충분하다는 것이다. 깊은 마취가 요구되는 경우, 예컨대 1 ~ 2 %의 이소 플루 란과 같은 흡입제가 적절할 수있다. </li><li> 동물이 얼음에 있지만, 에반스 블루 염료의 30 μL와 주사기를로드합니다. </li><li> 동물이 완전히 마취되면, 여전히 호흡하는 동안 얼음에 운동 부족에 의해 확인 현미경으로 이동합니다. 오른손 주입 들어, 오른쪽으로 동물의 총구에 직면한다. 이어 버드는 색인과 중간 손가락 (그림 1) 사이가되도록 이어 버드에 총구와 왼쪽 가운데 손가락 꼬리에 왼쪽 검지 손가락을 놓습니다. </li><li> 단지 피부가 조작 될 때 이동하는 표면 모세 혈관의 이어 버드로 전방 검사합니다. 이 모세관하지만, 대상이 아니다식별은 시간 정맥 식별을 위해 중요하다. 다음에 관계없이 피부의 위치에 고정되어 모세관 열등 어두운 그림자가 정맥을 찾습니다. 시간 정맥, 그림자가 나타납니다 지느러미에 복부 실행하고, 경정맥 (그림 1)에 공급한다. </li><li> 바늘 베벨 위로 시간적 정맥을 입력합니다. 올바르게 삽입하면 바늘 베벨 피부를 통해 혈액으로 채워 볼 수있다. 그런 다음 천천히 플런저를 우울하게하고 노트는 얼굴의 측면 아래 정맥의 창백. </li><li> 바늘이 주입 물 역류를 방지하기 위해 첨가 10-15 초 동안 정맥 내 유지하도록 허용. </li></ol> 3. 주입 후 <ol><li> 적절한 주입 한 후, 강아지는 거의 즉시 파란색 설정해야합니다. 바늘을 제거하고 혈액 응고 될 때까지 주사 부위에 면봉을 적용하는 부드러운 힘을 사용합니다. </li><li> 고통의 흔적이 강아지를 모니터링합니다. 강아지 2 ~ 3 분 복구하고 rewarm 허용, R강아지가 케이지에 반환하기 전에, 의식 수직 및 이동 때 ecognize. 컵 탐정의 장갑을 낀 손에 새끼 필요한 경우 복구에 도움이되는 적절한 온기를 제공합니다. 선택적으로, 주입 된 새끼 온난화 촉진하는 홈 케이지 하에서 가열 패드를 배치. 일반적으로 에반스 블루 염료를 주입 한 후, 새끼를 안락사. 시험 물질을 주입 할 때 염료를 포함하지 마십시오. </li><li> 다시 홈 케이지에 강아지를 놓고 강아지가 댐에 의해 reacceptance을 보장하기 위해 침구 및 / 또는 nestlet로 코팅되어 있는지 확인합니다. </li><li> 불임을 유지하기 위해 각각의 강아지에 대한 새 주사기와 면봉을 사용합니다. </li></ol>

<ol>
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저자가 공개하는 게 없다.

CNS 또는 몸 전체 에이전트의 혈관 배달 질환의 신생아 쥐 모델에서 어렵다. 설명 프로토콜은 정맥 최소한의 장비 요구 사항에 신생아 쥐에 솔루션을 관리 할 수​​있는 빠르고, 비교적 비 침습적 방법입니다. 시간적 얼굴 정맥을 육안으로 볼 수 있지만, 주사는 특히 unpracticed 인젝터, 현미경 및 광섬유 광원의 사용과 높은 정확도를 가질 수있다. 신생아 마우스에서 혈관 내 주사는 높은 성공률 <sup...

소아 질병의 쥐 모델에서 중추 신경계 (CNS)에 대한 치료제의 배달는 도전 할 부분이 남아있다. 모델 신생아 질병 상태를 직접 CNS 내에서 적절한 구조에 삽입하기 어려울 수 있으므로 소형 및 발달 미숙 한 것을 마우스. 치료제의 혈관 내 주입 CNS 1-5과 3,5-9 망막을 포함한 몸 전체에 세포, 약물, 또는 바이러스 벡터를 제공 할 수있는 비 침습적, 내약성 방법이다. 이전 출판물은 해부 현미경 11, 12 않고, transilluminator가 10, 11을 사용하여 시간적 얼굴 정맥 주입을 설명, 또는 두 사람을 요구하는 것은 (10)를 삽입 할 수 있습니다. 한 개인이 새끼를 주입 할 수 있으며, 광원은 시간적 정맥이 강아지를 만지고 고정 표면 외과 테이프의 필요성 또는 강아지의 부착을 제거하지 않은 볼 때문에 프로토콜에서 설명 주입 기술은 유리한transilluminator가 11 등. 마우스에 아데노 - 관련 바이러스 벡터 혈청 형 9 (AAV9)의 전달은 뇌와 척수 (그림 1)를 통해 신경 세포와 성상 세포에 강력한 표현을 생성합니다. 피상적 시간적 안면 정맥 내로 바이러스 벡터의 전달을 확실하게 혈관 신경 근육 질환 소아 척수 근육 위축 (SMA) 2,4,13,14을 치료하는 신생아 마우스에서 다양한 연구에 사용 된 궁극적으로 처리 된 마우스의 수명을 증가되었다. 신생아 쥐의 혈관 내 주입도 효과적으로 말초 신경계 및 말초 기관 (그림 2)을 대상으로합니다. AAV의 주입 후, 후근 신경절, 간, 심장, 골격근, 폐, 그리고 창자의 myenteric 신경 얼기의 전달은 1,3,6,7,15을 관찰되었다. CNS와 주변의 대폭적인 전달은 글로벌 표현을 필요로하는 질병에 대한 분사 이상이 방법을 만든다이러한 고셔병 (16) 및 기타 리소좀 저장 질환 17, 18, ​​판자의 질병과 관련된 신경 ceroid lipofuscinoses, 19 Bardet-Biedl 증후군, 유아 (20)에서 발생하는 증상의 발병에 유전 적 다중 시스템 장애로 형질 전환 유전자,. 혈관 신생 생쥐에 주입은 모델링 시스템 전체 소아 질환의 신규 한 방법으로 고려되어야한다. 이 기술은 큰 동물 모델 5,21로 번역 한 내용 및 혈관 내 주입 이미 치료제를 전달 임상 적으로 허용되는 방법으로 존재한다. 현재의 프로토콜은 출생 후 하루 2 사출 늦어도 피상적 인 시간적 얼굴 정맥을 통해 신생아 쥐에 에이전트를 도출하는 간단하고 효율적인 방법을 설명하는 하나의 완성 할 수있다, 개인 연습을 잘 새끼와 어미 모두 허용됩니다. 새끼 최소한의 고통을 경험하고 신속하게 복구 할 수 있습니다. ImportanTLY, 성공적인 주입은 관리 에이전트의 글로벌 배달 될 것입니다. 이 프로토콜은 바이러스 벡터, 신생아 쥐에 제약 제제 또는 세포의 전달에 적합하다.

<table><tbody><tr><td>Thinpro insulin syringe</td><td>Terumo</td><td>SS30M3009</td><td>3/10 cc, 3/8&quot; needle, 30 G, 1 per mouse</td></tr><tr><td>Evans blue dye</td><td>Sigma-Aldrich</td><td>E2129</td><td>Dilute to 1% with 1x Phosphate Buffered Saline</td></tr><tr><td>Cotton tipped applicators</td><td>Fisher Scientific</td><td>23-400-101</td><td></td></tr><tr><td>Fiber optic light source</td><td>Fisher Scientific</td><td>12-562-36</td><td></td></tr><tr><td>Dissecting microscope</td><td></td><td></td><td></td></tr></tbody></table>

intravenous injections in neonatal mice

정맥 주사는 치료제를 제공하기 위해 임상 적으로 적용 가능한 방법이다. 성인 설치류 큰 동물의 경우, 정맥 주사는 기술적으로 가능하고 일상이다. 그러나 일부 마우스 모델은 잠재적 인 치료의 관리가 어려워 빠른 진행과 질병의 조기 발병을 가질 수 있습니다. 시간 (또는 안면) 정맥은 마우스에서 이어 버드에 전방 및 해부 현미경을 사용하여 머리의 어느 한쪽에 생후 명확 처음 이틀 동안 표시된다. 이 윈도우 동안, 시간적 정맥 50 μl의 최대 양으로 주입 될 수있다. 주사는 안전하고 잘 새끼와 어미 모두 허용합니다. 전형적인 주입 과정은 강아지가 홈 케이지로 복귀 된 후 1-2 분 이내에 완료된다. 제 생후함으로써 정맥 시각화 어렵고 주입 과정은 기술적으로 불안정해진다. 이 기술은 아데노 - 관련 바이러스 (AAV) VECT의 전달을 위해 사용되어왔다차례로 선택한 바이러스 혈청 형에 따라 동물의 수명을 거의 몸 전체, 안정적인 형질 전환 유전자 발현을 제공 할 수 있습니다 ORS.

적절한 주입하는 동안, 정맥은 순간적으로 분명, 또는 희게을 설정해야합니다. 염료 주입하면 전체 강아지 초 ​​이내에 파란색 설정해야합니다. 부적절한 분사가 발생한 경우, 종종 주사 부위 밖으로 누출 될 머리 또는 목 주입 물에서 농축 피하 볼 러스있다. 부적절한 주사는 또한 목 주위에 멍이의 모양이 발생할 수 있습니다. (분사가 완전히 정맥에 배달되지 않은 IE) 일반적으로 더 아?...

Watch this Scientific Journal Video about Intravenous Injections in Neonatal Mice at JoVE.com

Intravenous Injections in Neonatal Mice

소아 질환의 동물 모델은 초기 발병과 적극적인 질병의 진행을 경험할 수 있습니다. 임상 어린 마우스 모델에 관련된 치료 전달 기술적 어려울 수있다. 이 프로토콜은 생후 두 생후 일 이내 신생 쥐 비 침습적 정맥 주입 방법을 설명한다.

신생아 마우스의 정맥 주사

Intravenous injection is a clinically applicable manner to deliver therapeutics. For adult rodents and larger animals, intravenous injections are ...

Research

JoVE Journal

Biology

58.2K 조회수.  Ohio State University.  소아 질환의 동물 모델은 초기 발병과 적극적인 질병의 진행을 경험할 수 있습니다. 임상 어린 마우스 모델에 관련된 치료 전달 기술적 어려울 수있다. 이 프로토콜은 생후 두 생후 일 이내 신생 쥐 비 침습적 정맥 주입 방법을 설명한다. 

비디오: Intravenous Injections in Neonatal Mice

Delivery of Therapeutic Agents Through Intracerebroventricular (ICV) and Intravenous (IV) Injection in Mice

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most often results in failure of potential therapeutics designed for neurodegenerative disorders 1,2. Neurodegenerative diseases such as Spinal Muscular Atrophy (SMA), in which the lower motor neurons are affected, can benefit greatly from introducing the therapeutic agents into the CNS. The purpose of this video is to demonstrate two different injection paradigms to deliver therapeutic materials into neonatal mice soon after birth. One of these methods is injecting directly into cerebral lateral ventricles (Intracerebroventricular) which results in delivery of materials into the CNS through the cerebrospinal fluid 3,4. The second method is a temporal vein injection (intravenous) that can introduce different therapeutics into the circulatory system, leading to systemic delivery including the CNS 5. Widespread transduction of the CNS is achievable if an appropriate viral vector and viral serotype is utilized. Visualization and utilization of the temporal vein for injection is feasible up to postnatal day 6. However, if the delivered material is intended to reach the CNS, these injections should take place while the blood brain barrier is more permeable due to its immature status, preferably prior to postnatal day 2. The fully developed blood brain barrier greatly limits the effectiveness of intravenous delivery. Both delivery systems are simple and effective once the surgical aptitude is achieved. They do not require any extensive surgical devices and can be performed by a single person. However, these techniques are not without challenges. The small size of postnatal day 2 pups and the subsequent small target areas can make the injections difficult to perform and initially challenging to replicate.

Delivery of Therapeutic Agents Through Intracerebroventricular ICV and Intravenous IV Injection in Mice

This article demonstrates two very different methods of injection: 1) into the brain (intracerebroventricular) and 2) systemic (intravenous) to introduce the therapeutic agents into the central nervous system of neonatal mice.

마우스에 Intracerebroventricular을 통해 치료 대리점 납품 (ICV)과 정맥 (IV) 사출

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most ...

연구

의학

Use of In vivo Imaging to Monitor the Progression of Experimental Mouse Cytomegalovirus Infection in Neonates

Human Cytomegalovirus (HCMV or HHV-5) is a life-threatening pathogen in immune-compromised individuals. Upon congenital or neonatal infection, the virus can infect and replicate in the developing brain, which may induce severe neurological damage, including deafness and mental retardation. Despite the potential severity of the symptoms, the therapeutic options are limited by the unavailability of a vaccine and the absence of a specific antiviral therapy. Furthermore, a precise description of the molecular events occurring during infection of the central nervous system (CNS) is still lacking since observations mostly derive from the autopsy of infected children. Several animal models, such as rhesus macaque CMV, have been developed and provided important insights into CMV pathogenesis in the CNS. However, despite its evolutionary proximity with humans, this model was limited by the intracranial inoculation procedure used to infect the animals and consistently induce CNS infection. Furthermore, ethical considerations have promoted the development of alternative models, among which neonatal infection of newborn mice with mouse cytomegalovirus (MCMV) has recently led to significant advances. For instance, it was reported that intraperitoneal injection of MCMV to Balb/c neonates leads to infection of neurons and glial cells in specific areas of the brain. These findings suggested that experimental inoculation of mice might recapitulate the deficits induced by HCMV infection in children. Nevertheless, a dynamic analysis of MCMV infection of neonates is difficult to perform because classical methodology requires the sacrifice of a significant number of animals at different time points to analyze the viral burden and/or immune-related parameters. To circumvent this bottleneck and to enable future investigations of rare mutant animals, we applied in vivo imaging technology to perform a time-course analysis of the viral dissemination in the brain upon peripheral injection of a recombinant MCMV expressing luciferase to C57Bl/6 neonates.

Use of In vivo Imaging to Monitor the Progression of Experimental Mouse Cytomegalovirus Infection in Neonates

Human Cytomegalovirus (HCMV) infection of neonates represents an important cause of mental retardation, yet the molecular events leading to virus-induced pathogenesis are still poorly understood. To investigate the dynamics of brain infection, we adapted whole-animal in vivo imaging to perform time-course analysis of neonates infected with a luciferase-recombinant virus.

사용<em&gt; 생체 내</em&gt; 이미징 신생아의 실험 쥐 거대 세포 바이러스 감염의 진행 상황을 모니터링 할 수

Human Cytomegalovirus (HCMV or HHV-5) is a life-threatening  pathogen in immune-compromised individuals. Upon congenital or neonatal  infection, the virus ...

면역학 및 감염병학

Intravenous and Intra-amniotic In Utero Transplantation in the Murine Model

In utero transplantation (IUT) is a unique and versatile mode of therapy that can be used to introduce stem cells, viral vectors, or any other substances early in the gestation. The rationale behind IUT for therapeutic purposes is based on the small size of the fetus, the fetal immunologic immaturity, the accessibility and proliferative nature of the fetal stem or progenitor cells, and the potential to treat a disease or the onset of symptoms prior to birth. Taking advantage of these normal developmental properties of the fetus, the delivery of hematopoietic stem cells (HSC) via an IUT has the potential to treat congenital hematologic disorders such as sickle cell disease, without the required myeloablative or immunosuppressive conditioning required for postnatal HSC transplants. Similarly, the accessibility of progenitor cells in multiple organs during development potentially allows for a more efficient targeting of stem/progenitor cells following an IUT of viral vectors for gene therapy or genome editing. Additionally, IUT can be used to study normal developmental processes including, but not limited to, the development of immunologic tolerance. The murine model provides a valuable and affordable means to understanding the potential and limitations of IUT prior to pre-clinical large animal studies and an eventual clinical application. Here, we describe a protocol for performing an IUT in the murine fetus through intravenous and intra-amniotic routes. This protocol has been used successfully to elucidate the necessary conditions and mechanisms behind in utero hematopoietic stem cell transplantation, tolerance induction, and in utero gene therapy.

Intravenous and Intra-amniotic In Utero Transplantation in the Murine Model

We describe a protocol for performing an in utero transplantation (IUT) through intravenous and intra-amniotic routes of injection in the murine model. This protocol can be used to introduce cells, viral vectors, and other substances into the unique immune-tolerant fetal environment.

정 맥 및 양수 내 Utero Murine 모델에 이식

In utero transplantation (IUT) is a unique and versatile mode of therapy that can be used to introduce stem cells, viral vectors, or any other substances ...

발생학

Intracerebroventricular Viral Injection of the Neonatal Mouse Brain for Persistent and Widespread Neuronal Transduction

With the pace of scientific advancement accelerating rapidly, new methods are needed for experimental neuroscience to quickly and easily manipulate gene expression in the mouse brain. Here we describe a technique first introduced by Passini and Wolfe for direct intracranial delivery of virally-encoded transgenes into the neonatal mouse brain. In its most basic form, the procedure requires only an ice bucket and a microliter syringe. However, the protocol can also be adapted for use with stereotaxic frames to improve consistency for researchers new to the technique. The method relies on the ability of adeno-associated virus (AAV) to move freely from the cerebral ventricles into the brain parenchyma while the ependymal lining is still immature during the first 12-24 hr after birth. Intraventricular injection of AAV at this age results in widespread transduction of neurons throughout the brain. Expression begins within days of injection and persists for the lifetime of the animal. Viral titer can be adjusted to control the density of transduced neurons, while co-expression of a fluorescent protein provides a vital label of transduced cells. With the rising availability of viral core facilities to provide both off-the-shelf, pre-packaged reagents and custom viral preparation, this approach offers a timely method for manipulating gene expression in the mouse brain that is fast, easy, and far less expensive than traditional germline engineering.

Here we demonstrate a technique for widespread neuronal transduction by intraventricular injection of adeno-associated virus into the neonatal mouse brain. This method provides a rapid and easy way to attain lifelong expression of virally-delivered transgenes.

영구와 광범위한 신경 세포의 형질 도입에 대한 신생아 마우스 뇌의 뇌실 바이러스 감염

With the pace of scientific advancement accelerating rapidly, new methods are needed for experimental neuroscience to quickly and easily manipulate gene ...

신경과학

Intracerebroventricular and Intravascular Injection of Viral Particles and Fluorescent Microbeads into the Neonatal Brain

In the study on the pathogenesis of viral encephalitis, the infection method is critical. The first of the two main infectious routes to the brain is the hematogenous route, which involves infection of the endothelial cells and pericytes of the brain. The second is the intracerebroventricular (ICV) route. Once within the central nervous system (CNS), viruses may spread to the subarachnoid space, meninges, and choroid plexus via the cerebrospinal fluid. In experimental models, the earliest stages of CNS viral distribution are not well characterized, and it is unclear whether only certain cells are initially infected. Here, we have analyzed the distribution of cytomegalovirus (CMV) particles during the acute phase of infection, termed primary viremia, following ICV or intravascular (IV) injection into the neonatal mouse brain. In the ICV injection model, 5 &#181;l of murine CMV (MCMV) or fluorescent microbeads were injected into the lateral ventricle at the midpoint between the ear and eye using a 10-&#181;l syringe with a 27 G needle. In the IV injection model, a 1-ml syringe with a 35 G needle was used. A transilluminator was used to visualize the superficial temporal (facial) vein of the neonatal mouse. We infused 50 &#181;l of MCMV or fluorescent microbeads into the superficial temporal vein. Brains were harvested at different time points post-injection. MCMV genomes were detected using the in situ hybridization method. Fluorescent microbeads or green fluorescent protein expressing recombinant MCMV particles were observed by fluorescent microscopy. These techniques can be applied to many other pathogens to investigate the pathogenesis of encephalitis.

Here, we describe a simple method of intracerebroventricular and intravascular injection of viral particles or fluorescent microbeads into the neonatal mouse brain. The localization pattern of the virus and nanoparticles could be detected by microscopic evaluation or by in situ hybridization.

신생아 뇌에 바이러스 성 입자와 형광 마이크로 비즈의 뇌실과 혈관 내 주입

In the study on the pathogenesis of viral encephalitis, the infection method is critical. The first of the two main infectious routes to the brain is the ...

Free-Hand Approach를 사용하여 마우스에서 뇌내 주사 수행

Free-Hand Intracerebroventricular Injections in Mice

The investigation of neuroendocrine systems often requires the delivery of drugs, viruses, or other experimental agents directly into the brains of mice. An intracerebroventricular (ICV) injection allows the widespread delivery of the experimental agent throughout the brain (particularly in the structures near the ventricles). Here, methods for making free-hand ICV injections in adult mice are described. By using visual and tactile landmarks on the heads of mice, injections into the lateral ventricles can be made rapidly and reliably. The injections are made with a glass syringe held in the experimenter&#39;s hand and placed at approximate distances from the landmarks. Thus, this technique does not require a stereotaxic frame. Furthermore, this technique requires only brief isoflurane anesthesia, which permits the subsequent assessment of mouse behavior and/or physiology in awake, freely behaving mice. Free-hand ICV injection is a powerful tool for the efficient delivery of experimental agents into the brains of living mice and can be combined with other techniques such as frequent blood sampling, neural circuit manipulation, or in vivo recording to investigate neuroendocrine processes.

저자 스포트라이트: 생식 신경 조절의 통찰력, 혁신 및 미래 방안 

Here, a simple and rapid approach for performing intracerebroventricular injections in mice using a free-hand approach (that is, without a stereotaxic device) is described.

The investigation of neuroendocrine systems often requires the delivery of drugs, viruses, or other experimental agents directly into the brains of mice. ...

A Neonatal Rodent Model of Retroorbital Vein Injection

Intravenous (iv) injection is the most used route of drug administration in neonates in the clinical setting. Therefore, retroorbital vein injection is an important method for compound administration in research, where successful proof-of-concept studies can progress into much-needed neonatal clinical trials. Most intravenous studies in neonatal rodents use the superficial temporal/facial vein. However, retroorbital injection becomes unreliable in neonatal rodents older than 2 days after the skin darkens and the vein is no longer visible. In the present protocol, we describe the retroorbital injection of the venous sinus in both the neonatal mouse and rat at ages when the superficial temporal vein is no longer visible, but the eyes have not opened yet. Eye-opening facilitates retro-orbital injection by enabling the researcher to clearly see that they are not perforating the eye when inserting the needle. We demonstrate that this technique can be performed in a reliable and reproducible manner without adverse effects. Additionally, we show that it can be used in many studies, such as administering compounds to study neonatal brain injury.

This protocol aims to demonstrate a reproducible venous administration route that can be used in rats and mice throughout the neonatal period. This procedure is important for preclinical rodent studies that wish to mirror drug administration in neonatal care units primarily using intravenous administration.

후안와 정맥 주입의 신생아 설치류 모델

Intravenous (iv) injection is the most used route of drug administration in neonates in the clinical setting. Therefore, retroorbital vein injection is an ...

신생아 마우스에서 척수강내 주사를 전달하기 위한 포괄적인 프로토콜

Intrathecal Injection of Newborn Mouse for Genome Editing and Drug Delivery

Intrathecal injection is a commonly employed procedure in both pediatric and adult clinics, serving as an effective means to administer medications and treatments. By directly delivering medications and treatments into the cerebrospinal fluid of the central nervous system, this method achieves higher localized drug concentrations while reducing systemic side-effects compared to other routes such as intravenous, subcutaneous, or intramuscular injections. Its importance extends beyond clinical settings, as intrathecal injection plays a vital role in preclinical studies focused on treating neurogenetic disorders in rodents and other large animals, including non-human primates. However, despite its widespread application, intrathecal injection in young, particularly neonatal pups, poses significant technical challenges due to their small size and fragile nature. Successful and reliable administration of intrathecal injections in newborn mice requires meticulous attention to detail and careful consideration of various factors. Thus, there is a crucial need for a standardized protocol that not only provides instructions but also highlights key technical considerations and good laboratory practices to ensure procedural consistency, as well as the safety and welfare of the animals.
To address this unmet need, we present a detailed and comprehensive protocol for performing intrathecal injections specifically in newborn pups on postnatal day 1 (P1). By following the step-by-step instructions, researchers can confidently perform intrathecal injections in neonatal pups, enabling the accurate delivery of drugs, antisense oligos, and viruses for gene replacement or genome editing-based treatments. Furthermore, the importance of adhering to good laboratory practices is emphasized to maintain the well-being of animals and ensure reliable experimental outcomes. This protocol aims to address the technical challenges associated with intrathecal injections in neonatal mice, ultimately facilitating advances in the field of neurogenetic research that aims to develop potential therapeutic interventions.

저자 스포트라이트: 척수강내 주사 – 신생아 마우스 뇌에서 유전자 편집의 효능을 테스트하기 위한 효율적이고 신뢰할 수 있는 전달 방법

The present protocol outlines step-by-step instructions for performing intrathecal injections in neonatal mice for gene editing and drug delivery.

게놈 편집 및 약물 전달을 위한 신생아 마우스의 척수강내 주사

Intrathecal injection is a commonly employed procedure in both pediatric and adult clinics, serving as an effective means to administer medications and ...

신생아 마우스의 정맥 주사

요약

더 많은 비디오 탐색

이 비디오의 챕터

마우스에 Intracerebroventricular을 통해 치료 대리점 납품 (ICV)과 정맥 (IV) 사출

사용

정 맥 및 양수 내 Utero Murine 모델에 이식

영구와 광범위한 신경 세포의 형질 도입에 대한 신생아 마우스 뇌의 뇌실 바이러스 감염

신생아 뇌에 바이러스 성 입자와 형광 마이크로 비즈의 뇌실과 혈관 내 주입

Free-Hand Intracerebroventricular Injections in Mice

Free-Hand Intracerebroventricular Injections in Mice

Free-Hand Intracerebroventricular Injections in Mice

후안와 정맥 주입의 신생아 설치류 모델

게놈 편집 및 약물 전달을 위한 신생아 마우스의 척수강내 주사