Os autores gostariam de reconhecer o NINDS, FightSMA e Famílias de SMA para apoio financeiro. SEGL é suportado por formação NINDS concessão # 5T32NS077984-02.

Todos os procedimentos listados no protocolo foram aprovados Instituto para uso animal e comissão Care (IACUC) da Universidade Estadual de Ohio. 1. Preparação da área de trabalho <ol><li> Reunir gelo triturado, para anestesiar os filhotes de rato, uma gaiola vazia para segregar a represa da ninhada, um microscópio de dissecação, uma fonte de luz que pode ser posicionado a um ângulo em relação à injecção (utilização de uma fonte de luz em um 90 ° &#160; ângulo em relação ao local da injecção obscurece a veia), uma superfície limpa para colocar o animal para a injecção, cotonetes, 3/10 cc seringa de insulina com 3/8 &quot;de 30 L de agulha (um por animal) e 1% de corante azul de Evans ( feita com) solução salina tamponada com fosfato (PBS) para a formação. </li><li> Retire a barragem de uma gaiola separada ao manipular os filhotes. </li></ol> 2. Injecção Procedimento <ol><li> Coloque um único filhote diretamente no gelo molhado por 30-60 seg para anestesiar o animal. Não deixe tele animais no gelo por muito tempo devido a um risco de complicações associadas à hipotermia relacionada, incluindo a fibrilação ventricular, hipóxia tecidual e acidose metabólica. NOTA: A nossa experiência é que 30-60 segundos é suficiente para abrandar os movimentos filhote de permitir a injecção. Se a anestesia mais profunda é necessária, inalantes, tais como 1-2% de isoflurano pode ser apropriada. </li><li> Enquanto o animal é em gelo, carregar a seringa com 30 ul de corante azul de Evans. </li><li> Quando o animal é completamente anestesiados, confirmada pela falta de movimento no gelo enquanto ainda em respirar, movê-lo sob o microscópio. Para uma injecção destro, enfrentar o focinho do animal para a direita. Coloque o dedo indicador esquerdo no focinho eo caudal dedo médio esquerdo à orelha broto para que a orelha broto é entre o indicador eo dedo médio (Figura 1). </li><li> Examine apenas anterior à orelha broto de um capilar superficial que se move quando a pele é manipulado. Este capilar não é o alvo, no entantoidentificação é importante para a identificação da veia temporal. Em seguida, localize um escuro, sombrio veia inferior ao capilar que permanece fixo, independentemente da posição da pele. A veia temporal, aparece sombreada, corre para ventral dorsal, e alimenta-se na veia jugular (Figura 1). </li><li> Digite a veia temporal com a agulha de bisel para cima. Se inserido corretamente, é possível visualizar os bisel da agulha se enchem de sangue através da pele. Em seguida, pressionar o êmbolo devagar e com nota branqueamento da veia para o lado do rosto. </li><li> Permitir que a agulha para permanecer dentro da veia para um agregado de 10-15 seg para evitar o refluxo do injectant. </li></ol> 3. Pós-injecção <ol><li> Após uma injecção adequada, o filhote deve ficar azul quase que imediatamente. Remover a agulha e usar a força suave para aplicar um cotonete para o local de injecção até que os coágulos de sangue. </li><li> Monitorar o filhote para sinais de sofrimento. Permitir que o filhote 2-3 min para recuperar e reaquecer, recognize quando o filhote está consciente, de pé e em movimento, antes de voltar para a jaula. Taça dos filhotes nas mãos enluvadas do investigador para fornecer calor adequado para ajudar na recuperação, se necessário. Como alternativa, coloque uma almofada de aquecimento sob a gaiola para facilitar o aquecimento do filhote injetado. Geralmente depois de uma injeção de corante azul de Evans, eutanásia filhotes. Não incluem corante ao injetar o material de teste. </li><li> Coloque o filhote de volta para a gaiola e garantir que o filhote é revestido com roupa de cama e / ou nestlet para garantir reacceptance pela barragem. </li><li> Use uma seringa nova e cotonete para cada cachorro para manter a esterilidade. </li></ol>

<ol>
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Os autores não têm nada a revelar.

A entrega intravascular de agentes para o SNC ou por todo o corpo é difícil em modelos murinos de doença neonatal. O protocolo descrito é uma maneira rápida, relativamente não-invasivo para administrar por via intravenosa soluções em camundongos neonatal com requisitos mínimos de equipamento. Embora a cara veia temporal, podem ser vistos a olho nu, as injecções podem ter maior precisão com o uso do microscópio e fonte luminosa de fibra óptica, especialmente para um injector sem prática. Injeções intrave...

A entrega de agentes terapêuticos ao sistema nervoso central (SNC) em modelos murinos de doença pediátrica continua a ser um desafio. Ratos que modelo estados de doença recém-nascidos são subdimensionados e developmentally imaturo, e, portanto, pode ser difícil de injectar directamente nas estruturas apropriadas dentro do SNC. Injeção intravascular de agentes terapêuticos é um método não-invasivo, bem tolerado para entregar células, drogas ou vetores virais para todo o corpo, incluindo o SNC 1-5 e 3,5-9 retina. Publicações anteriores descrevem injeção veia rosto temporal, utilizando um transiluminador 10,11, sem um microscópio de dissecação 11,12, ou exigindo duas pessoas para injetar 10. A técnica de injeção descritos neste protocolo é vantajoso porque um único indivíduo pode injetar filhotes, ea fonte de luz para ver a veia temporal, não está tocando o filhote, eliminando a necessidade de fita cirúrgica ou a fixação de um filhote de cachorro para uma superfície fixatal como um transiluminador 11. Entrega de adeno-associado vector viral serótipo 9 (AAV9) em ratinhos produz expressão robusta em neurónios e astrócitos em todo o cérebro e na medula espinal (Figura 1). A entrega intravascular de vectores virais na veia facial temporal superficial foi usado de forma fiável em vários estudos em ratinhos neonatais para tratar a doença neuromuscular pediátrica atrofia muscular espinal (SMA) e, finalmente, 2,4,13,14 aumentou o tempo de vida dos ratinhos tratados. Injecção intravascular de ratinhos neonatais também tem como alvo de forma eficaz o sistema nervoso periférico e órgãos periféricos (Figura 2). Após a injecção de AAV, a transdução de gânglios da raiz dorsal, fígado, coração, músculo esquelético, o pulmão, e do plexo mientérico do intestino foi observada 1,3,6,7,15. Transdução generalizada do sistema nervoso central e periferia torna este método de injeção ideal para doenças que requerem expressão global deum transgene, tais como a doença de 16 e outro de depósito lisossômico doenças de Gaucher, doença de 17,18 Batten e lipofuscinoses Ceróides neuronais relacionados, 19 e síndrome de Bardet-Biedl, uma doença genética do sistema múltiplo com início dos sintomas que ocorrem na infância 20. Injecção intravascular em ratinhos neonatais, também deve ser considerado como um novo método de doenças pediátricas em todo o sistema de modelagem. Esta técnica tem sido traduzido para animais de grande porte 5,21 e injeção intravascular já existe como um método clinicamente aceitável de entregar terapêutica. O protocolo atual descreve um método simples e eficiente de agentes para ratos neonatais entregar através da veia superficial rosto temporal, o mais tardar até dia pós-natal 2. Injeção pode ser completado por um único, praticada individual e é bem tolerado por ambos os filhotes e as barragens. Filhotes experimentar angústia mínima e recuperar rapidamente. Importantly, injecção bem sucedido resultará na entrega global do agente administrado. Este protocolo é apropriado para a entrega de vectores virais, agentes farmacêuticos ou de células para ratinhos recém-nascidos.

<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

A injeção intravenosa é uma forma clinicamente aplicável para entregar terapêutica. Para roedores adultos e animais maiores, injeções intravenosas são tecnicamente viáveis ​​e de rotina. No entanto, alguns modelos de mouse pode ter início precoce da doença com uma progressão rápida que torna a administração de terapias potenciais difícil. A veia temporal (ou facial) é apenas anterior ao botão da orelha em ratinhos e é claramente visível para os dois primeiros dias após o nascimento em cada lado da cabeça, usando um microscópio de dissecação. Durante esta janela, a veia temporal, podem ser injectados com volumes de até 50 ul. A injecção é seguro e bem tolerado por ambas as crias e as barragens. Um procedimento típico de injecção está concluída dentro de 1-2 min, após o que o filhote é devolvida à gaiola de origem. No terceiro dia pós-natal a veia é difícil de visualizar e do procedimento de injeção se torna tecnicamente confiável. Esta técnica tem sido utilizada para a entrega de vírus adeno-associado (AAV) vectors, que por sua vez pode fornecer quase todo o corpo, a expressão do transgene estável durante a vida do animal, dependendo do serotipo viral escolhido.

Durante uma injecção adequada, a veia deve momentaneamente virar clara, ou branquear. Se injetar tingir todo o filhote deve ficar azul em segundos. Se uma injecção indevida tenha ocorrido, há muitas vezes um bolus subcutâneo concentrada na cabeça ou pescoço e injectant pode verter para fora do local de injecção. Injecções impróprios também pode resultar no aparecimento de nódoas negras em torno da garganta. Os filhotes que recebem injeções subcutâneas (ou seja, a injeção não foi totalmente ...

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

Intravenous Injections in Neonatal Mice

Modelos animais de doença pediátrica pode experimentar início precoce e progressão da doença agressiva. Clinicamente entrega terapia relevantes para jovens modelos de mouse pode ser tecnicamente difícil. Este protocolo descreve um método de injecção intravenosa não-invasiva para a ratinhos recém-nascidos nos dois primeiros dias pós-natais de vida.

Injeções intravenosas em Neonatal Ratos

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

58.1K visualizações.  Ohio State University.  Modelos animais de doença pediátrica pode experimentar início precoce e progressão da doença agressiva. Clinicamente entrega terapia relevantes para jovens modelos de mouse pode ser tecnicamente difícil. Este protocolo descreve um método de injecção intravenosa não-invasiva para a ratinhos recém-nascidos nos dois primeiros dias pós-natais de vida. 

Vídeo: 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.

Entrega de agentes terapêuticos Através intracerebroventricular (ICV) e injecção intravenosa (IV) em ratos

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 ...

Medicina

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.

Utilização de<em&gt; In vivo</em&gt; Imagem para monitorar a progressão da infecção pelo citomegalovírus Experimental Rato em recém-nascidos

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

Imunologia e Infecção

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.

Intravenosa e Intra-amniótica no Utero transplante no modelo murino

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 ...

Biologia do Desenvolvimento

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.

Injection Viral intracerebroventricular do Neonatal do rato do cérebro para a transdução neuronal persistente e generalizada

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

Neurociência

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.

Intracerebroventricular e intravascular A injeção de Viral partículas e fluorescentes microesferas no cérebro Neonatal

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 ...

Realização de injeção intracerebroventricular em camundongos usando uma abordagem à mão livre

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.

Autor em destaque: insights, inovações e caminhos futuros no neurocontrole reprodutivo 

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.

Injeções intracerebroventriculares à mão livre em camundongos

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.

Modelo Neonato de Injeção de Veias Retroorbitárias em Roedores

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

Um protocolo abrangente para administração de injeções intratecais em camundongos neonatais

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.

Autor em destaque: Injeção intratecal - um método de entrega eficiente e confiável para testar a eficácia da edição de genes em cérebros de camundongos neonatais

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

Injeção Intratecal de Camundongos Recém-Nascidos para Edição do Genoma e Liberação de Fármacos

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

Injeções intravenosas em Neonatal Ratos

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