Name: intraductal delivery to the rabbit mammary gland
Uploaded: 2017-03-09T13:00:00.000+00:00
Duration: 6 min 30 s
Description: Watch this Scientific Journal Video about Intraductal Delivery to the Rabbit Mammary Gland at JoVE.com

The authors acknowledge support from a Translational Breast Cancer Research Grant (14-60-26-BROC to AB) from the Breast Cancer Research Foundation and the American Association for Cancer Research.

Procedimentos usando indivíduos animais foram aprovados pelo Comitê de Cuidado e Uso Institucional Animal da Universidade do Texas em Austin. 1. Pré-operatório Preparação <ol><li> Registe o peso do corpo de cada coelho. Tal como acontece com todos os estudos pré-clínicos, monitorar os pesos dos animais regularmente para avaliar a toxicidade potencial. </li><li> Antes de se anestesiar o coelho, encher um tubo de 50 mL cónico com gel de ultra-sons comercialmente disponível e de centrifugação a 500 xg durante 30 s; não deve haver bolhas visíveis no gel após a conclusão da centrifugação. </li><li> Administrar glicopirrolato por via subcutânea numa dose de 0,1 mg / kg e acepromazina por via intramuscular numa dose de 0,75 mg / kg. Aguarde 15-20 minutos para o sedativo para entrar em vigor durante o acompanhamento de sinais e comportamentos vitais. NOTA: O glicopirrolato é um agente anti-colinérgico que impede bradicardia e reduz respiratórias e secreções gastrointestinais. Acepromazina é um sedativo que serve como um premedicatipor anestesia. </li><li> Administrar 35 mg / kg de cetamina e 5 mg / kg de xilazina por via subcutânea como um anestésico. No entanto, como o operador torna-se mais experiente e pode realizar a entrega intraductal mais rapidamente, diminuir as doses de droga a 15 mg / kg de cetamina e 3 mg / kg de xilazina por via subcutânea; isto irá diminuir o tempo de anestesia e o tempo necessário para o animal a recuperar da anestesia. </li><li> Verificar e documentar a frequência cardíaca, SPO 2, temperatura, frequência respiratória, e cor membrana mucosa a cada 15 minutos. Aplique lubrificante olho em ambos os olhos. NOTA: Apenas pessoal que tenha recebido uma formação adequada e tenham sido aprovados pelo IACUC da sua instituição deve administrar ou controlar a anestesia. A utilização de um monitor de anestesia veterinária pode auxiliar na aquisição de sinais vitais, é recomendada. O monitor de anestesia veterinária, no entanto, não substitui a necessidade de verificar manualmente o animal a cada 15 minutos. Veja as instruções do fabricante fou o uso adequado de um monitor de veterinária. </li><li> Verifique se o início da anestesia por uma pitada toe suave; o coelho deve ser não-responsivo antes de continuar. </li><li> Cuidadosamente raspar o abdome caudal do coelho na área em torno do terceiro e quarto pares de tetas inguinais. </li><li> Com a maioria dos cabelos removidos, aplicar um creme de depilação over-the-counter à área rapada. Retire o creme de 10 min após a aplicação usar toalhas de papel húmidas molhadas com água morna. NOTA: pele de coelho é muito frágil / sensível. O creme de depilação deve ser usado para não mais do que 10 min. De facto, é mais seguro para tentar um &quot;ponto de teste&quot; em 5 min e apenas deixar no local já não se teste indica um tempo mais longo é necessário período. </li><li> Limpe a área com compressas de gaze embebido em álcool para limpar o local da injeção. </li><li> Coloque o coelho no seu dorso em uma calha em forma de v forrada com um cobertor água morna recirculação e uma almofada absorvente. </li></ol> 2. Preparation do agente de contraste <ol><li> Reconstituir o reagente não-alvo de contraste de acordo com as instruções do fabricante. Pipeta cima e para baixo suavemente para misturar. NOTA: O agente de contraste utilizado na presente protocolo é estável à temperatura ambiente durante 4-6 h depois da reconstituição. Agite suavemente o frasco entre cada recuperação. NOTA: O volume da solução requerida irá depender do número de condutas a ser injectada. O coelho tem aberturas 4 ductais por tetina, e 0,2 mL de solução é suficiente para preencher uma árvore ductal de um adulto coelho branco da Nova Zelândia (Oryctolaguscuniculus). Assim, um volume total de 0,8 mL pode ser entregue a uma glândula mamaria. </li></ol> 3. Entrega Intraductal <ol><li> Localize as tetas apropriadas a serem injetados; os 3º e 4º pares de tetas inguinais são recomendados, pois eles são facilmente visualizados quando o animal é posicionado sobre seu dorso. </li><li> Carga 0,2 ml de solução salina estéril 0,9% emuma seringa de tuberculina de 1 ml de Luer-lock com uma agulha 22 G. Descarte a agulha G 22 uma vez que a solução salina é na seringa e substituí-lo com uma agulha de calibre 25 estéril. Limpe cuidadosamente a área com 85% de álcool isopropílico em uma gaze. </li><li> Com o bisel da agulha para cima e a seringa paralela ao corpo do animal, inserir o bisel da agulha para o lado da tetina e injectar lentamente 0,1-0,2 mL de solução salina; isso vai permitir uma melhor visualização das aberturas ductal. </li><li> Carregar 0,2 ml de solução de injecção para uma seringa de tuberculina de 1 ml de bloqueio Luer. </li><li> Segure a tetina suavemente com o polegar eo dedo indicador e levante-o ligeiramente para posicioná-la para a injeção intraductal; uma lupa de ocular wearable pode ajudar a visualizar as aberturas ductal. </li><li> Enquanto se mantém a posição elevada do teto, canular cuidadosamente a conduta de interesse usando uma agulha de ponta romba 25 G. </li><li> Após punção, gire suavemente a seringa Luer-lock no cubo da franco-tagulha da infusão ip até travar no lugar. </li><li> Levantar a tetina e injectar a solução lentamente para minimizar o dano potencial causado pelo movimento rápido do fluido no interior da conduta; em nenhum momento deve haver resistência ao injetar a solução. </li></ol> 4. Ecografia <ol><li> Aplique uma quantidade generosa de gel de ultra-som centrifugado para a pele da área de interesse. Certifique-se de que não existem bolhas no gel, como estes irão comprometer a qualidade da imagem. </li><li> Defina a profundidade da imagem a 6 mm. Posicionar o transdutor de 21 MHz em contacto com o gel e digitalizar a área de interesse em modo-B. Observar o meio de contraste na região digitalizada, inclusive a abertura ductal e ao longo da conduta. NOTA: Estas definições foram desenvolvidos para uso com uma máquina de ultra-som fotoacústica específico. Consulte a tabela de materiais para mais detalhes. Pode ser necessário ajustar a potência de transmissão e a profundidade da imagem de outros sistemas de imagem para optimizar mamáriavisualização glândula. </li><li> Remover o gel de ultra-som da pele do animal com uma almofada de gaze. </li></ol> 5. Cuidados no pós-operatório <ol><li> Observar o local da injecção: não deve haver sinais de trauma para a região da tetina ou para o tecido circundante, e inchaço na área circundante da tetina provavelmente indica uma injecção de almofada de gordura mamaria em vez de uma injecção intraductal bem sucedida. </li><li> Coloque o coelho numa posição esternal. Se necessário, dar 0,2 mg / kg de ioimbina por via intravenosa na veia marginal da orelha; isto irá reverter o efeito da xilazina e permitir que o animal a recuperar mais rapidamente da anestesia. </li><li> Monitorar o coelho a cada 15 min durante todo o período de recuperação até que o animal está alerta, ágil e mantém uma posição esternal. NOTA: Este procedimento não deve resultar em danos nos tecidos ou inchaço. Se vermelhidão ou inchaço são observados, administrar uma dose de meloxicam de 0,1-0,2 mg / kg, PO, uma vez que o animal é alerta e capazde tomar a medicação por via oral. Entre em contato com o pessoal veterinário da instituição para obter mais orientações. </li></ol>

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The authors have nothing to disclose.

Este método de entrega intraductal para a glândula mamaria de coelho pode ser utilizada para reagentes de contraste para ultrassons e muitas outras soluções aquosas, incluindo corantes vitais e terapêutica. Estudos anteriores demonstraram a entrega intraductal de hormônios 17, 18, 19. Em modelos de roedores, a entrega intraductal de ácidos nucleicos 8, quimioterápicos 6,</s...

<html> A entrega intraductal de agentes terapêuticos tem sido estudada em modelos de roedores e em início de carreira testes em humanos 3, 4, 5, 6, 11, 12. A Fase I recente estudo demonstrou a segurança ea viabilidade de carboplatina intraductal ou intraductal doxorrubicina lipossomal peguilada em mulheres que aguardam mastectomia para o tratamento de câncer invasivo 2. Protocolos anteriores para entrega intraductal foram desenvolvidos para rato e mamárias de ratos glândulas 6, 7, 8, 9. Para fins de investigação, as injecções de células de tumor intraductal e a entrega de vector lentiviral de oncogenes também foram realizados em modelos de roedoresref &quot;&gt; 13, 14, 15, 16. No entanto, um ideal em modelo vivo do processo de entrega intraductal deve permitir o desenvolvimento de novas classes de compostos terapêuticos e facilitar a avaliação pré-clínica. diferenças anatômicas entre roedores e humanos têm complicado a tradução destes estudos. Ao contrário dos ratos, em que cada conduta termina a uma tetina separada, da mama humano consiste de 5 a 9 ductais sistemas independentes, cada um com uma abertura separada terminando na tetina. Coelho glândulas mamárias abrigar quatro sistemas ductais independentes, cada um separadamente, acessível através de um de quatro orifícios em uma única tetina. Um modelo de coelho corresponda melhor a anatomia humana e permite o estudo da entrega da droga intraductal em um contexto mais relevante. Aqui, usamos duas técnicas para avaliar a prestação intraductal. A co-administração de umcorante vital permite a visualização através da pele e fornece uma confirmação simples e rápida do método. Para algumas aplicações, o mapeamento de resolução mais elevado dos ductos podem ser preferidos. Nós apresentamos aqui um protocolo para ultra-sonografia das condutas através da entrega intraductal de um reagente não-alvo contraste.

<table><tbody><tr><td>MicroMarker non-targeted contrast reagent</td><td>VisualSonics</td><td>VS-11694</td><td></td></tr><tr><td>Luer Lock 1 mL Syringes</td><td>BD</td><td>309628</td><td></td></tr><tr><td>Glycopyrrolate 0.2 mg/mL</td><td>Wedgewood Compounding Pharmacy</td><td>GLYCOP-INJ013VC</td><td>6 month shelf life, supply may be limited.&amp;nbsp;</td></tr><tr><td>Atropine Sulfate 0.5 mg/mL</td><td>Animal Health International</td><td>15320764</td><td>If glycopyrrolate is unavailable. Not to be combined with glycopyrrolate.</td></tr><tr><td>Ketamine HCL 100 mg/mL</td><td>Animal Health International</td><td>21250699</td><td>&lt;a href=&quot;http://www.animalhealthinternational.com/&quot;&gt;http://www.animalhealthinternational.com/&lt;/a&gt;</td></tr><tr><td>Acepromazine 10 mg/mL</td><td>Animal Health International</td><td>17640541</td><td></td></tr><tr><td>Xylazine 20 mg/mL</td><td>Animal Health International</td><td>20101547</td><td></td></tr><tr><td>Yohimbine 0.2 mg/mL</td><td>Animal Health International</td><td>14588965</td><td></td></tr><tr><td>Hair Removing Cream</td><td>Veet</td><td></td><td>Sensitive skin solution. Available through local retailers.</td></tr><tr><td>Blunt tip infusion needles</td><td>Sai Infusion Technology</td><td>B14-50</td><td>&lt;a href=&quot;http://www.sai-infusion.com/collections/blunt-needles&quot;&gt;http://www.sai-infusion.com/collections/blunt-needles&lt;/a&gt;</td></tr><tr><td>Veterinary Pulse Oximeter</td><td>EdanUSA</td><td>VE-H100B</td><td>&lt;a href=&quot;http://www.edanusa.com/Product/VE-H100B-Veterinary-Pulse-Oximeter.html&quot;&gt;http://www.edanusa.com/Product/VE-H100B-Veterinary-Pulse-Oximeter.html&lt;/a&gt;</td></tr><tr><td>Warm Water Pump</td><td>Gaymar</td><td>TP700</td><td></td></tr><tr><td>Warm Water Blanket</td><td>Animal Health International</td><td>21232696</td><td>Maxi-Therm Lite Warming Pads</td></tr><tr><td>Ultrasound system</td><td>VisualSonics</td><td>Vevo 2100</td><td></td></tr></tbody></table>

intraductal delivery to the rabbit mammary gland

Intraductais tratamentos localizados para o cancro da mama oferecem vantagens potenciais, incluindo a entrega eficiente para o tumor e reduziu a toxicidade sistémica e efeitos adversos 1, 2, 3, 4, 5, 6, 7. No entanto, vários desafios permanecem antes que esses tratamentos podem ser aplicados de forma mais ampla. O desenvolvimento e validação de terapêuticas intraductais em um modelo animal apropriado facilitar o desenvolvimento de estratégias terapêuticas intraductais para os pacientes. Enquanto a glândula mamaria do rato tem sido amplamente utilizada como um sistema modelo de desenvolvimento mamário e a tumorigénese, a anatomia é distinta da glândula humano. Um modelo animal maior, tal como o coelho, pode servir como modelo para uma melhor estrutura de glândula mamaria e no desenvolvimento terapêutico intraductal. em cOntrast para ratos, em que dez árvores ductal são espacialmente distribuídos ao longo do eixo do corpo, cada uma terminando em uma teta separada, a glândula mamária de coelho mais se assemelha a glândula humana, com vários sistemas sobrepostos ductal que a saída através de aberturas separadas em uma teta. Aqui, apresentamos métodos minimamente invasivos para a entrega de reagentes diretamente no ducto mamário coelho e para visualização do próprio parto com a ultra-sonografia de alta resolução.

Aqui, mostra-se que a entrega de reagentes intraductal contraste com as condutas mamarias de um coelho pode ser alcançado sem trauma para o tecido (Figura 2). Em coelhos, quatro sistemas ductais separadas convergem para uma tetina e, assim, podem ser acedidos individualmente e fotografada utilizando este método. aberturas ductal individuais são facilmente visualizado; Observe a marcação de uma segunda abertura ductal adjacente à conduta canulada na Figura 2...

Watch this Scientific Journal Video about Intraductal Delivery to the Rabbit Mammary Gland at JoVE.com

Intraductal Delivery to the Rabbit Mammary Gland

Here, we describe a technique for the localized delivery of reagents to the rabbit mammary gland via an intraductal injection. In addition, we describe a protocol for visualization and the confirmation of delivery by high-resolution ultrasound imaging of contrast agents.

Entrega Intraductal ao Coelho glândula mamária

Localized intraductal treatments for breast cancer offer potential advantages, including efficient delivery to the tumor and reduced systemic toxicity and ...

intraductal-delivery-to-the-rabbit-mammary-gland

Research

JoVE Journal

Bioengineering

9.3K visualizações.  The University of Texas at Austin. Here, we describe a technique for the localized delivery of reagents to the rabbit mammary gland via an intraductal injection. In addition, we describe a protocol for visualization and the confirmation of delivery by high-resolution ultrasound imaging of contrast agents. 

Vídeo: Intraductal Delivery to the Rabbit Mammary Gland

Minimally Invasive Embryo Transfer and Embryo Vitrification at the Optimal Embryo Stage in Rabbit Model

Assisted reproductive techniques (ARTs), such as in vitro embryo culture or embryo cryopreservation, affect natural development patterns with perinatal and postnatal consequences. To ensure the innocuousness of ART applications, studies in animal models are necessary. In addition, as a last step, embryo development studies require evaluation of their capacity to develop full-term healthy offspring. Here, embryo transfer to the uterus is indispensable to perform any ARTs-related experiment.
The rabbit has been used as a model organism to study mammalian reproduction for over a century. In addition to its phylogenetic proximity to the human species and its small size and low maintenance cost, it has important reproductive characteristics such as induced ovulation, a chronology of early embryonic development similar to humans and a short gestation that allow us to study the consequences of ART application easily. Moreover, ARTs (such as intracytoplasmic sperm injection, embryo culture, or cryopreservation) are applied with suitable efficiency in this species.
Using the laparoscopic embryo transfer technique and the cryopreservation protocol presented in this article, we describe 1) how to transfer embryos through an easy, minimally invasive technique and 2) an effective protocol for long-term storage of rabbit embryos to provide time-flexible logistical capacities and the ability to transport the sample. The outcomes obtained after transferring rabbit embryos at different developmental stages indicate that morula is the ideal stage for rabbit embryo recovery and transfer. Thus, an oviductal embryo transfer is required, justifying the surgical procedure. Furthermore, rabbit morulae are successfully vitrified and laparoscopically transferred, proving the effectiveness of the described techniques.

Assisted reproductive techniques (ARTs) are in continuous evaluation to improve outcomes and reduce the associated risks. This manuscript describes a minimally invasive embryo transfer procedure with an efficient cryopreservation protocol that allows the use of rabbits as an ideal animal model of human reproduction.

Transferência de embriões minimamente invasivos e vitrificação embrionária na fase de embrião ideal em modelo de coelho

Assisted reproductive techniques (ARTs), such as in vitro embryo culture or embryo cryopreservation, affect natural development patterns with perinatal ...

Biologia do Desenvolvimento

Intraductal Delivery and X-ray Visualization of Ethanol-Based Ablative Solution for Prevention and Local Treatment of Breast Cancer in Mouse Models

Breast cancer is the most prevalent cancer and the second-leading cause of cancer-related death for women in the USA. For high-risk women, prophylactic mastectomy is the most effective primary prevention strategy. Prophylactic mastectomy is an aggressive surgical procedure that completely removes the mammary epithelial cells from which breast cancer arises along with the surrounding tissue. We seek to develop a minimally invasive intraductal procedure as an alternative to prophylactic mastectomy to locally ablate the mammary epithelial cells before they can become malignant. We and others have developed an intraductal delivery procedure to reach and treat these epithelial cells in rodent models of breast cancer. While the mouse mammary gland with a single non-anastomosed ductal tree opening at the nipple has a much less complex and tortuous architecture than the human breast, chemically induced and genetically engineered mouse models of breast cancer are valuable to produce proof-of-concept studies of new preventative strategies. Here, we describe a procedure for intraductal delivery of an ethanol-based ablative solution containing micro-CT/X-ray tantalum-based contrast agent within the mouse mammary ductal tree for the therapeutic purpose of primary prevention of breast cancer. Intraductal delivery of aqueous reagents (e.g., cytotoxic compounds, siRNAs, AdCre) has been previously described in mouse models. Thus, we focus our protocol description on methodological modifications and unique experimental considerations for optimizing delivery of ethanol, for minimizing local and systemic side effects of ethanol administration, and for in vivo visualization of ductal tree filling via micro-CT/fluoroscopy imaging. Visualization of the ductal tree immediately after injection of a contrast-containing solution allows for confirmation of complete filling or unsuccessful outcomes such as underfilling or overfilling. This procedure can be applied for delivery and imaging of other ablative compounds aimed at either preventing tumor formation or locally treating early-stage tumors accessible via the ductal tree.

A method of intraductal injection of reagents for an ethanol-based ablative solution to the mouse mammary ductal tree for in vivo imaging and breast cancer prevention is described. Injection directly into the nipple opening allows for targeting mammary epithelial cells with minimal collateral tissue damage.

Entrega intraduto e visualização de raios-X da solução ablativa baseada em etanol para prevenção e tratamento local do câncer de mama em modelos de camundongos

Breast cancer is the most prevalent cancer and the second-leading cause of cancer-related death for women in the USA. For high-risk women, prophylactic ...

Pesquisa do câncer

X-Ray Visualization of Intraductal Ethanol-Based Ablative Treatment for Prevention of Breast Cancer in Rat Models

There are still a limited number of primary interventions for prevention of breast cancer. For women at a high risk of developing breast cancer, the most effective intervention is prophylactic mastectomy. This is a drastic surgical procedure in which the mammary epithelial cells that can give rise to breast cancer are completely removed along with the surrounding tissue. The goal of this protocol is to demonstrate the feasibility of a minimally invasive intraductal procedure that could become a new primary intervention for breast cancer prevention. This local procedure would preferentially ablate mammary epithelial cells before they can become malignant. Intraductal methods to deliver solutions directly to these epithelial cells in rodent models of breast cancer have been developed at Michigan State University and elsewhere. The rat mammary gland consists of a single ductal tree that has a simpler and more linear architecture compared to the human breast. However, chemically induced rat models of breast cancer offer valuable tools for proof-of-concept studies of new preventive interventions and scalability from mouse models to humans. Here, a procedure for intraductal delivery of an ethanol-based ablative solution containing tantalum oxide nanoparticles as X-ray contrast agent and ethyl cellulose as gelling agent into the rat mammary ductal tree is described. Delivery of aqueous reagents (e.g., cytotoxic compounds, siRNAs, AdCre) by intraductal injection has been described previously in mouse and rat models. This protocol description emphasizes methodological changes and steps that pertain uniquely to delivering an ablative solution, formulation consideration to minimize local and systemic side effects of the ablative solution, and X-ray imaging for in vivo assessment of ductal tree filling. Fluoroscopy and micro-CT techniques enable to determine the success of ablative solution delivery and the extent of ductal tree filling thanks to compatibility with the tantalum-containing contrast agent.

A procedure for the delivery of a chemical ablative solution to the rat mammary ductal tree for image-guided preventive treatment of breast cancer is described. Mammary epithelial cells can be targeted with minimal collateral tissue damage through cannulation directly into the nipple opening and intraductal infusion of a 70% ethanol-based ablative solution.

Visualização de Raios-X do Tratamento Ablativo Intraductal à Base de Etanol para Prevenção do Câncer de Mama em Modelos de Rato

There are still a limited number of primary interventions for prevention of breast cancer. For women at a high risk of developing breast cancer, the most ...

In Vivo Gene Delivery into Mouse Mammary Epithelial Cells Through Mammary Intraductal Injection

Mouse mammary glands comprise ductal trees, which are lined by epithelial cells and have one opening at the tip of each nipple. The epithelial cells play a major role in mammary gland function and are the origin of most mammary tumors. Introducing genes of interest into mouse mammary epithelial cells is a critical step in evaluating gene function in epithelial cells and generating mouse mammary tumor models. This goal can be accomplished through the intraductal injection of a viral vector carrying the genes of interest into the mouse mammary ductal tree. The injected virus subsequently infects mammary epithelial cells, bringing in the genes of interest. The viral vector can be lentiviral, retroviral, adenoviral, or adenovirus-associated viral (AAV). This study demonstrates how a gene of interest is delivered into mammary epithelial cells through mouse mammary intraductal injection of a viral vector. A lentivirus carrying GFP is used to show stable expression of a delivered gene, and a retrovirus carrying Erbb2 (HER2/Neu) is used to demonstrate oncogene-induced atypical hyperplastic lesions and mammary tumors.

In Vivo Gene Delivery into Mouse Mammary Epithelial Cells Through Mammary Intraductal Injection

The present protocol describes intraductal injection of viral vectors via the teat to deliver genes of interest into the mammary epithelial cells.

In vivo Entrega de Genes em Células Epiteliais Mamárias de Camundongos Através de Injeção Intraductal Mamária

Mouse mammary glands comprise ductal trees, which are lined by epithelial cells and have one opening at the tip of each nipple. The epithelial cells play ...

Intranasal Immunization and Milk Collection in Studies of Maternal Immunization in New Zealand White Rabbits (Oryctolagus cuniculus)

Due to similarities in placentation and antibody transfer with humans, rabbits are an excellent model of maternal immunization. Additional advantages of this research model are the ease of breeding and sample collection, relatively short gestation period, and large litter sizes. Commonly assessed routes of immunization include subcutaneous, intramuscular, intranasal, and intradermal. Nonterminal sample collection for the chronological detection of the immunologic responses to these immunizations include the collection of blood, from both dams and kits, and milk from the lactating does. In this article, we will demonstrate techniques our lab has utilized in studies of maternal immunization in New Zealand White rabbits (Oryctolagus cuniculus), including intranasal immunization and milk collection.

Intranasal Immunization and Milk Collection in Studies of Maternal Immunization in New Zealand White Rabbits Oryctolagus cuniculus

This article describes and demonstrates the administration of intranasal vaccines and the collection of milk from lactating rabbits (Oryctolagus cuniculus) as a means to assess mucosal immunity in a translationally appropriate model of maternal immunization.

Imunização Intranasal e Coleta de Leite em Estudos de Imunização Materna em Coelhos Brancos da Nova Zelândia (Oryctolagus cuniculus)

Due to similarities in placentation and antibody transfer with humans, rabbits are an excellent model of maternal immunization. Additional advantages of ...

Imunologia e Infecção

Intraductal Injection for Localized Drug Delivery to the Mouse Mammary Gland

Herein we describe a protocol to deliver various reagents to the mouse mammary gland via intraductal injections. Localized drug delivery and knock-down of genes within the mammary epithelium has been difficult to achieve due to the lack of appropriate targeting molecules that are independent of developmental stages such as pregnancy and lactation. Herein, we describe a technique for localized delivery of reagents to the mammary gland at any stage in adulthood via intraductal injection into the nipples of mice. The injections can be performed on live mice, under anesthesia, and allow for a non-invasive and localized drug delivery to the mammary gland. Furthermore, the injections can be repeated over several months without damaging the nipple. Vital dyes such as Evans Blue are very helpful to learn the technique. Upon intraductal injection of the blue dye, the entire ductal tree becomes visible to the eye. Furthermore, fluorescently labeled reagents also allow for visualization and distribution within the mammary gland. This technique is adaptable for a variety of compounds including siRNA, chemotherapeutic agents, and small molecules.

A protocol for the non-invasive intraductal delivery of aqueous reagents to the mouse mammary gland is described. The method takes advantage of localized injection into the nipples of mammary glands targeting mammary ducts specifically. This technique is adaptable for a variety of compounds including siRNA, chemotherapeutic agents and small molecules.

Injeção Intraductal para localizada Drug Delivery ao Rato glândula mamária

Herein we describe a protocol to deliver various reagents to the mouse mammary gland via intraductal injections. Localized drug delivery and knock-down of ...

Biologia

Simplifying Transcutaneous Intratracheal Drug Delivery in the Newborn Preterm Rabbit

Intratracheal (IT) drug delivery allows the direct delivery of pharmaceutical substances to the lung, maximizing potential pulmonary benefit and minimizing systemic drug exposure. The transcutaneous technique is simple and allows for the IT delivery of substances to the lung of prematurely born rabbits shortly after birth. Newborn pups are anesthetized with inhaled Isoflurane before being placed in a supine position with the neck extended. The larynx is identified and stabilized before transcutaneous placement of a 26-gauge (G) catheter into the trachea. Following catheterization of the trachea, a 30 G blunt needle attached to a Hamilton syringe is introduced into the IT catheter and is used for delivering a precise volume into the trachea during spontaneous respiration. After the IT injection is completed, the needle and catheter are withdrawn, and the pup is allowed to recover from anesthesia. Transcutaneous IT injection delivers a large proportion of the injected substance to the lung, with the majority remaining in the lung 3 hours after the intervention. The injections are well tolerated from the day of birth and can be repeated for multiple consecutive days without influencing survival. This technique can be used to investigate the effect of pharmaceutical agents on lung development and in the prevention of neonatal lung injury in preterm rabbits.

Transcutaneous intratracheal injection allows for effective intrapulmonary drug delivery during spontaneous respiration. Single and multiple injections are well tolerated with no effect on survival. The technique is simple to perform and can examine the effect of substances on lung development and the prevention of lung injury in newborn rabbits.

Intratracheal (IT) drug delivery allows the direct delivery of pharmaceutical substances to the lung, maximizing potential pulmonary benefit and ...

Medicina

Pregnancy and Nursing Management for Embryo-Transferred and Genetically Modified Rabbits

With the advancement of scientific research, the demand for gene-edited rabbit models is increasing. However, there are limited pregnancy and feeding management systems for gene-edited rabbits, leading to low survival rates among gene-edited rabbits prepared by many inexperienced researchers. Therefore, proper guidance is essential. This article summarizes the pregnancy and feeding practices for genetically modified rabbits developed in the author's laboratory and outlines a set of fundamental processes. These include pregnancy diagnosis, antenatal care, midwifery, assisted breastfeeding, weaning, and other procedures, along with the rescue and care of weak newborn rabbits. Compared to the traditional natural childbirth and nurturing methods used in rabbit farms, this approach involves more refined management, requiring additional time and effort but significantly increasing the survival rate of suckling rabbits. The methods described in this article are suitable for most laboratory breeding scenarios involving gene-edited or embryo-transferred rabbits and provide a straightforward and effective reference for other researchers.

This protocol describes a pregnancy and feeding management technique for embryo-transferred and genetically modified rabbits, aimed at reducing newborn rabbit mortality and enhancing the preparation efficiency of gene-edited rabbits.

Manejo da Gravidez e Amamentação para Coelhos Transferidos com Embriões e Geneticamente Modificados

With the advancement of scientific research, the demand for gene-edited rabbit models is increasing. However, there are limited pregnancy and feeding ...

Intraductal Injection: Delivering Injection Mix into the Ducts of the Mouse Mammary Gland

Source: Xiang, D., et al. <a href="https://www.jove.com/v/59502/modeling-breast-cancer-via-an-intraductal-injection-cre-expressing">Modeling Breast Cancer via an Intraductal Injection of Cre-expressing Adenovirus into the Mouse Mammary Gland.</a> J. Vis. Exp (2019).
This video describes an intraductal injection technique where we deliver injection mix into the ducts of mouse mammary glands.

Injeção intraductal: Aplicação da mistura de injeção nos ductos da glândula mamária do camundongo

Source: Xiang, D., et al. Modeling Breast Cancer via an Intraductal Injection of Cre-expressing Adenovirus into the Mouse Mammary Gland. J. Vis. Exp ...

JoVE Encyclopedia of Experiments

Câncer de mama

Procedimentos e Técnicas

Rabbit Intraductal Injection: Localized Delivery of Solution of Interest into the Rabbit Mammary Gland

Source: Clark et. al., <a href="https://www.jove.com/v/55209/intraductal-delivery-to-the-rabbit-mammary-gland">Intraductal Delivery to the Rabbit Mammary Gland.</a> J. Vis. Exp. (2017).
In this video we describe a technique of administering a solution of interest into the mammary duct of a rabbit and confirm the delivery by high-resolution ultrasound imaging of contrast agents.

Injeção intraductal em coelhos: entrega localizada de solução de interesse na glândula mamária de coelho

Source: Clark et. al., Intraductal Delivery to the Rabbit Mammary Gland. J. Vis. Exp. (2017).
In this video we describe a technique of administering a ...