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.

Le procedure che utilizzano soggetti animali sono stati approvati dal Comitato di Cura e uso istituzionale degli animali della University of Texas di Austin. 1. Preparazione preoperatoria <ol><li> Registrare il peso corporeo di ogni coniglio. Come per tutti gli studi preclinici, monitorare i pesi degli animali regolarmente per valutare la potenziale tossicità. </li><li> Prima di anestetizzare il coniglio, riempire una provetta da 50 ml con gel per ultrasuoni disponibili in commercio e far girare a 500 xg per 30 s; ci dovrebbero essere visibili bolle nel gel al termine della centrifugazione. </li><li> Somministrare glicopirrolato per via sottocutanea alla dose di 0,1 mg / kg e acepromazine intramuscolare alla dose di 0,75 mg / kg. Attendere 15-20 minuti per il sedativo abbia effetto, mentre il monitoraggio dei segni vitali e il comportamento. NOTA: Glicopirrolato è un agente anti-colinergico che impedisce bradicardia e riduce respiratorie e GI secrezioni. Acepromazine è un sedativo che funge da premedication per l&#39;anestesia. </li><li> Somministrare 35 mg / kg di ketamina e 5 mg / kg di xylazina sottocutanea come anestetico. Tuttavia, come l&#39;operatore diventa più esperto e può compiere la consegna intraduttale più rapidamente, diminuire i dosaggi farmaceutiche a 15 mg / kg di ketamina e 3 mg / kg di xylazina sottocutanea; questo ridurrà il tempo di anestesia e il tempo richiesto per l&#39;animale per recuperare da anestesia. </li><li> Controllare e documentare la frequenza cardiaca, SPO 2, temperatura, frequenza respiratoria, e il colore membrana mucosa ogni 15 min. Applicare il lubrificante occhio in entrambi gli occhi. NOTA: Solo il personale che hanno subito la formazione adeguata e sono stati approvati dalla IACUC della loro istituzione dovrebbe amministrare o monitorare l&#39;anestesia. L&#39;uso di un monitor anestesia veterinario può aiutare ad acquisire segni vitali ed è consigliato. Il monitor anestesia veterinaria, tuttavia, non sostituisce la necessità di controllare manualmente l&#39;animale ogni 15 min. Vedere le istruzioni del produttore Fo l&#39;uso corretto di un monitor veterinaria. </li><li> Verificare l&#39;insorgenza di anestesia da un pinch punta dolce; il coniglio deve essere non reattivi prima di continuare. </li><li> radersi con cura l&#39;addome caudale del coniglio nella zona attorno al terzo e quarto coppie di mammelle inguinali. </li><li> Con la maggior parte dei capelli rimossi, applicare una crema depilatoria over-the-counter all&#39;area rasata. Rimuovere il min crema 10 dopo l&#39;applicazione usando tovaglioli di carta umidi bagnate con acqua tiepida. NOTA: la pelle di coniglio è molto fragile / sensibile. La crema depilatoria deve essere utilizzato per non più di 10 min. In realtà, è più sicuro provare un &quot;punto di prova&quot; a 5 min e lasciare solo il più a lungo se posto test indica un tempo più lungo è necessario periodo. </li><li> Pulire l&#39;area con tamponi di garza imbevuta di alcol per pulire il sito di iniezione. </li><li> Posizionare il coniglio sul suo dorso in un recipiente a forma di v foderato con un ricircolo coperta acqua calda e un tampone assorbente. </li></ol> 2. Preparation del mezzo di contrasto <ol><li> Ricostituire il reagente non mirati contrasto in base alle istruzioni del produttore. Pipetta su e giù delicatamente per miscelare. NOTA: L&#39;agente di contrasto utilizzato in questo protocollo è stabile a temperatura ambiente per 4 - 6 h dopo la ricostituzione. oscillare delicatamente il flaconcino tra ogni recupero. NOTA: Il volume di soluzione necessario dipende dal numero di condotti da iniettare. Il coniglio ha 4 aperture duttale per tettarella, e 0,2 ml di soluzione è sufficiente a riempire un albero duttale di un adulto Nuova Zelanda White Rabbit (Oryctolaguscuniculus). Così, un volume totale di 0,8 ml può essere consegnato ad una ghiandola mammaria. </li></ol> 3. La consegna intraduttale <ol><li> Individuare i capezzoli appropriate da iniettare; i 3 ° e 4 ° coppie di mammelle inguinali sono raccomandati, poiché essi vengono facilmente visualizzate quando l&#39;animale è posizionato sul suo dorso. </li><li> Carico 0,2 mL di sterile salina allo 0,9% inuna siringa da tubercolina da 1 ml Luer-serratura con un ago 22 G. Smaltire l&#39;ago G 22 una volta che la soluzione salina è nella siringa e sostituirlo con un ago calibro 25 sterile. Pulire delicatamente l&#39;area con 85% alcol isopropilico ed un tampone di garza. </li><li> Con la smussatura dell&#39;ago e la siringa parallelo al corpo dell&#39;animale, inserire la smussatura dell&#39;ago nel lato del capezzolo e iniettare lentamente 0,1-0,2 mL di soluzione fisiologica; questo permetterà per una migliore visualizzazione delle aperture duttale. </li><li> Carico 0,2 ml di soluzione per l&#39;iniezione in una siringa da tubercolina 1 ml Luer-lock. </li><li> Mantenere la tettarella delicatamente con il pollice e l&#39;indice e sollevarla leggermente per posizionarlo per l&#39;iniezione intraduttale; una lente di ingrandimento oculare indossabile può essere di aiuto nel visualizzare le aperture duttale. </li><li> Pur mantenendo la posizione sollevata del capezzolo, cannulate accuratamente il condotto di interesse utilizzando un ago smussato punta 25 G. </li><li> Dopo l&#39;incannulamento, torcere delicatamente la siringa luer-lock sul mozzo della smussato-tip ago infusione fino a quando non si blocca in posizione. </li><li> Sollevare la tettarella e iniettare la soluzione lentamente per ridurre al minimo i potenziali danni causati dal rapido movimento fluido all&#39;interno della condotta; in nessun momento ci dovrebbe essere la resistenza quando si inietta la soluzione. </li></ol> 4. Ultrasound Imaging <ol><li> Applicare una quantità di gel ultrasuoni centrifugato alla pelle della zona di interesse. Verificare che non vi siano bolle nel gel, in quanto questi saranno compromettere la qualità dell&#39;immagine. </li><li> Impostare la profondità di imaging a 6 mm. Posizionare il trasduttore 21 MHz in contatto con il gel e la scansione dell&#39;area di interesse in B-mode. Osservare il mezzo di contrasto nella regione scansionata tra cui l&#39;apertura duttale e nel condotto. NOTA: Queste impostazioni sono stati sviluppati per l&#39;utilizzo con una specifica macchina ad ultrasuoni fotoacustica. Fare riferimento alla tabella dei materiali per maggiori dettagli. Può essere necessario regolare la potenza di trasmissione e la profondità di imaging di altri sistemi di imaging per ottimizzare mammariavisualizzazione della ghiandola. </li><li> Rimuovere il gel per ultrasuoni dalla pelle dell&#39;animale con un tampone di garza. </li></ol> 5. Cura postoperatoria <ol><li> Osservare il sito di iniezione: non ci dovrebbero essere i segni di un trauma alla regione del capezzolo o al tessuto circostante, e gonfiore nella zona circostante la tettarella probabilmente indica una mammaria iniezione cuscinetto adiposo, piuttosto che una iniezione di intraduttale successo. </li><li> Posizionare il coniglio in una posizione sternale. Se necessario, dare 0,2 mg / kg per via endovenosa di yohimbina alla vena marginale dell&#39;orecchio; ciò invertire l&#39;effetto della xilazina e permettere all&#39;animale di recuperare più rapidamente da anestesia. </li><li> Monitorare il coniglio ogni 15 minuti per tutto il periodo di recupero fino a quando l&#39;animale è vigile, reattivo, e mantiene una posizione sternale. NOTA: Questa procedura non dovrebbe causare danni ai tessuti o gonfiore. Se rossore o gonfiore si osservano, somministrare una dose di meloxicam 0.1-0.2 mg / kg PO una volta che l&#39;animale è vigile e in gradodi assumere farmaci per via orale. Contattare personale veterinario dell&#39;istituto per ulteriori indicazioni. </li></ol>

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The authors have nothing to disclose.

Questo modo di somministrazione intraduttale alla ghiandola mammaria coniglio può essere utilizzato per i reagenti di contrasto ecografici e molte altre soluzioni acquose, compresi coloranti vitali e terapeutica. Studi precedenti hanno dimostrato la consegna intraduttale di ormoni 17, 18, 19. Nei modelli di roditori, la consegna intraduttale di acidi nucleici 8, chemioterapici 6,<...

<html> La consegna intraduttale di agenti terapeutici è stata studiata in modelli di roditori e in fase iniziale sperimentazione umana 3, 4, 5, 6, 11, 12. Un recente studio di fase I ha dimostrato la sicurezza e la fattibilità di carboplatino intraduttale o intraduttale doxorubicina liposomiale pegilata in donne in attesa di mastectomia per il trattamento del cancro invasivo 2. Precedenti protocolli per la consegna intraduttale sono stati sviluppati per topo e ratto mammarie ghiandole 6, 7, 8, 9. Per scopi di ricerca, iniezioni di cellule tumorali intraduttale e la consegna lentivirale vettore di oncogeni sono stati anche eseguiti in modelli di roditoriref &quot;&gt; 13, 14, 15, 16. Tuttavia, un ideale modello in vivo del processo di consegna intraduttale dovrebbe consentire lo sviluppo di nuove classi di composti terapeutici e facilitare la valutazione preclinica. differenze anatomiche tra i roditori e gli esseri umani hanno complicato la traduzione di questi studi. Diversamente topi, in cui ciascun condotto termina in un capezzolo separata, il seno umano è costituito da 5 a 9 sistemi duttali indipendenti, ognuno con un&#39;apertura separata termina al capezzolo. Coniglio ghiandole mammarie porto quattro sistemi duttale indipendenti, ciascuna separatamente, accessibile attraverso uno dei quattro orifizi in una sola tettarella. Un modello di coniglio più si avvicina al anatomia umana e consente lo studio di drug delivery intraduttale in un contesto più rilevante. Qui, usiamo due tecniche per valutare la consegna intraduttale. La co-somministrazione di uncolorante vitale permette la visualizzazione attraverso la pelle e fornisce una conferma semplice e rapida del metodo. Per alcune applicazioni, la mappatura risoluzione maggiore dei condotti può essere preferito. Presentiamo qui un protocollo per l&#39;imaging ad ultrasuoni dei condotti attraverso la consegna intraduttale di un reagente non mirato contrasto.

<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

Trattamenti intraduttale localizzati per il cancro al seno offrono potenziali vantaggi, compresa la lettura efficiente al tumore e ridotta tossicità sistemica e effetti avversi 1, 2, 3, 4, 5, 6, 7. Tuttavia, rimangono molte sfide prima di questi trattamenti possono essere applicati più ampiamente. Lo sviluppo e la validazione di terapie intraduttale in un modello animale adeguato facilitano lo sviluppo di strategie terapeutiche per i pazienti intraduttale. Mentre la ghiandola mammaria topo è stato ampiamente utilizzato come sistema modello di sviluppo mammario e tumorigenesi, l&#39;anatomia è distinta dalla ghiandola umana. Un modello animale più grande, come il coniglio, può servire come un modello migliore per la struttura della ghiandola mammaria e sviluppo terapeutico intraduttale. in contrast a topi, in cui dieci alberi dotto sono spazialmente distribuiti lungo l&#39;asse del corpo, ciascuna terminante in un capezzolo a parte, la ghiandola mammaria di coniglio assomiglia più da vicino la ghiandola umana, con più sistemi duttale sovrapposti che escono attraverso le aperture separate in una tettarella. Qui, vi presentiamo i metodi minimamente invasive per la fornitura di reagenti direttamente nel condotto mammaria coniglio e per la visualizzazione della stessa consegna con ecografia ad alta risoluzione.

Qui mostriamo che la consegna intraduttale di reagenti contrasto ai dotti mammari di un coniglio può essere raggiunta senza traumi al tessuto (Figura 2). Nei conigli, quattro sistemi duttale separati convergono in una sola tettarella e, quindi, possono essere consultati e ripreso individualmente con questo metodo. Aperture individuali duttale sono facilmente visualizzati; notare la marcatura di una seconda apertura duttale adiacente al condotto cannulata in Figu...

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.

Consegna intraduttale al Coniglio ghiandola mammaria

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 Views.  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. 

Video: 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.

Trasferimento embrionale minimamente invasivo e vetrificazione embrionale nella fase embrionale ottimale nel modello Rabbit

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

Ricerca

Biologia dello sviluppo

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.

Somministrazione intraduttale e visualizzazione a raggi X della soluzione ablativa a base di etanolo per la prevenzione e il trattamento locale del cancro al seno in modelli murini

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 ...

Ricerca sul cancro

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.

Visualizzazione a raggi X del trattamento ablativo intraduttale a base di etanolo per la prevenzione del cancro al seno in modelli di ratto

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 Consegna genica nelle cellule epiteliali mammarie di topo attraverso l'iniezione intraduttale mammaria

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.

Immunizzazione intranasale e raccolta del latte negli studi sull'immunizzazione materna nei conigli bianchi della Nuova Zelanda(Oryctolagus cuniculus)

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

Immunologia e infezioni

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.

Iniezione intraduttale per Localizzato Drug Delivery per il mouse ghiandola mammaria

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.

Gestione della gravidanza e dell'allattamento per conigli trasferiti in embrioni e geneticamente modificati

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.

Iniezione intraduttale: erogazione di una miscela per iniezione nei dotti della ghiandola mammaria del topo

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

Breast Cancer

Procedures and techniques

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.

Iniezione intraduttale di coniglio: somministrazione localizzata di soluzione di interesse nella ghiandola mammaria del coniglio

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 ...