Breanne Cuddington holds a fellowship from the Canadian Breast Cancer Foundation. This work was sponsored by operating grants from the Cancer Research Society and the Canadian Cancer Society Research Institute (formerly the Canadian Breast Cancer Research Alliance). We thank Ann Tollefson (Saint Louis University School of Medicine) for LCRT cells and Dr. Kathleen Delaney and Marion Corrick for technical assistance with cotton rat housing and sedation.

<ol>
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The authors acknowledge there are no financial conflicts of interest related to this research.

Cotton rats are highly excitable and have a strong flight response. Therefore, special care should be taken to minimize any undue stress on the animal. The cage setup described will allow for safe and easy capture of the animals, with the placement of the enrichment tube being of the utmost importance. When setting up cages, ensure that the enrichment tubes meet the size and shape requirements, and are placed in proper orientation in the cage. It is also important to ensure that any technicians who might be aiding in ani...

וירוסי Oncolytic (OV) לשכפל באופן סלקטיבי בתאי גידול על ידי ניצול הבדלים ביוכימיים בין תאים נורמלים וסרטניים 1,2. ישנם שני סוגים של OVS: אלה שאינם דורשים מוטציה להשיג oncolysis סלקטיבית, המכונים וירוסי wild-type כמתרחשים באופן טבעי ואלה אשר חייבים להיות מהונדסים כדי להשיג oncolysis סלקטיבית. האוסף של מוטציות בתוך סוג גידול נתון קובע את אופי יתרון הצמיחה סלקטיבית על פני תאים נורמלים לOV 2. הבטיחות והתועלת של OVS באמצעות הודגמה בניסויים קליניים 3-7. למרות התקדמות בתחום virotherapy oncolytic קיימים פערים בין תוצאות פרה-קליניות וקליניות, המצביע על כך יש צורך במודלים טובים יותר להעריך את היעילות אנטי-הסרטנית של OVS. סוג הווירוס ההרפס שור 1 (BHV-1) הוא חבר של משפחת הרפס, ותת-משפחת Alphaherpesviridae. BHV-1 initiates השור מורכב מחלה בדרכי הנשימה בבקר, מפגין במגוון רחב של תסמינים דומים 8,9 קר רע. BHV-1 נקשר קולטנים התקשרות וכניסה בשימוש על ידי HSV-1, כגון סולפט וnectin-1 10 heparan. עם זאת, הוא נקשר CD155 במקום של nectin-2 10. יש BHV-1 טווח מארח צר מאוד כך שהוא אינו יכול להיכנס ביעילות ובליזום שכפול בתאים עכבריים רגילים והפכו את 3,4,10. זה הופך את השימוש במודלים עכבריים קונבנציונליים בעייתי. קיבולת oncolytic של BHV-1 כבר הוכיחה במבחנה 11,12. BHV-1 הוכח ליזום שכפול ובלהרוג תאים סרטניים אנושיים ממגוון רחב של מקורות היסטולוגית, כוללים תאי סרטן השד וסרטן שד ייזום תאים 11,12. עם זאת, היכולת אנטי-הסרטנית של BHV-1 חייבת להיות מוערכת in vivo בהקשר של מארח עם מערכת חיסון תקין. האדם Adenovirus (Ad), שליש 57 זנים אל מזוהים, בדרך כלל גורם למחלה בדרכי הנשימה בבני אדם. וקטורי Ad Oncolytic נבדקו ליעילות אנטי-הסרטנית שלהם עם כמה קידום לניסויים קליניים 13-15. למרות נתונים פרה-קליניים מבטיחים, תוצאות קליניות נפלו קצרות של ציפיות. מודלים xenograft גידול אנושי משמשים בדרך כלל כדי לחקור את היעילות אנטי-הסרטנית של וקטורי מודעות, למרות שהם מפגינים נחלשו מערכת חיסונית לווירוס 16,17. יתר על כן, מודלים עכבריים syngeneic הם אוֹסְרָנִי לזיהום מודעות, מה שהופכים את ההערכה של תגובות חיסוני מארח תוך שימוש במודלים אלה מעשיים 17,18. המערכת החיסונית המארח זוהתה כמנגנון המשפיע ביותר על ידי שOVS להוציא מוות של תאים סרטניים 19. תגובות אנטי-סרטניות בין tolerized ואנטיגן גידולים הקשורים אינו tolerized דגמים (TAA) שונים ויכולות להשפיע על ההצלחה של טיפול OV מאוד. OV KM100 HSV-1 (ICP0 n212VP16 בשנת 1814 20) 20,21 עורר נסיגה של גידול ב -80% מעכברי נושאי גידול במודל סרטן שד אנטיגן Polyoma התיכון T העכברי 22. עם זאת, בדגמים / neu HER-2, היעילות אנטי-הסרטנית של KM100 נעה בין 20% רגרסיה מלאה בעכברי syngeneic וקיפאון בגידול מהונדס, HER2-tolerized עכברים. יחד נתונים אלה מדגישים את החשיבות של הערכה מלאה OVS באמצעות מודלים של בעלי חיים בצורה הטובה ביותר לשחזר את נוף החיסון האנושי כדי להבין מה תכונות שתקבענה את ההצלחה טיפולית. החולדה הכותנה (hispidus Sigmodon), ילידי צפון אמריקה והדרום, היא נפוצה ביותר כמודל של זיהום בנגיף syncytial נשימה (כפי שנסקר ב5). חולדות כותנה משמשות גם במחקר חיסון נגד BHV-1 כפי שהם לשחזר את הפתולוגיה הקשורים למחלות בדרכי הנשימה שור מורכבות 6,23. יתר על כן, זיהום BHV-1 של חולדות כותנההוא חיסוני, גרימה רירית מתמשכים ותגובות חיסוניים מערכתיות 6,23-25. שורות תאים כבר נגזרו fibrosarcoma וosteosarcomas של בלוטת החלב (LCRT) והעצם (CCRT וVCRT), בהתאמה 26 ספונטניים. חולדות כותנה היו בשימוש כדי להעריך את יעילות in vivo של וקטורי Ad oncolytic כפי שהם רגישים לזיהום מודעות ולהפגין פתולוגיה דומה לבני האדם 27-29. השימוש במודלי מדוכאי חיסון להערכה טרום הקלינית של OVS הם לא רק פחות מעיד על תגובה הקלינית לטיפול, אבל הם אינם לוקחים בחשבון את תפקידה של מערכת החיסון בvirotherapy oncolytic 30,31. לכן, syngeneic והמודלים של עכברים כותנה-tolerized גידול של סרטן השד וסרטן עצמות הם מודלים רלוונטיים שבו להעריך את היעילות פרה-הקלינית של OVS, כגון BHV-1 ומודעות שלא ניתן ללמוד באמצעות מודלים עכבריים קונבנציונליים.

<table><tbody><tr><td>Dulbecco&amp;rsquo;s modified Eagle&amp;rsquo;s medium&amp;nbsp;</td><td>Gibco</td><td>11965-092</td><td>May use any brand&amp;nbsp;</td></tr><tr><td>1X Phosphate Buffered Saline&amp;nbsp;</td><td></td><td></td><td>Can prepare in lab, filter to sterilize</td></tr><tr><td>200 mM L-glutamine</td><td>Gibco</td><td>25030164</td><td>May use any brand</td></tr><tr><td>100x Antibiotic-Antimycotic&amp;nbsp;</td><td>Gibco</td><td>15240-062</td><td>May use any brand</td></tr><tr><td>Fetal bovine serum</td><td>Quality Biological Inc.</td><td>110-001-101HI</td><td>May use any brand</td></tr><tr><td>T-150cm&lt;sup&gt;2&lt;/sup&gt; tissue culture flask</td><td>Fisher Scientific</td><td>14-826-80</td><td>May use any brand</td></tr><tr><td>1X TypLE Express</td><td>Life Technologies</td><td>12604-013</td><td></td></tr><tr><td>12-well cell culture plate, flat bottom</td><td>Fisher Scientific</td><td>08-772-29</td><td>May use any brand, must be tissue culture treated</td></tr><tr><td>alamarBlue</td><td>Life Technologies</td><td>DAL1025</td><td>May use an alternative reagent for determination of cell viability</td></tr><tr><td>8640 Teklad 22/5 Rodent diet</td><td>Harlan&amp;nbsp;</td><td>8640</td><td></td></tr><tr><td>1/8&amp;rdquo; corncob rodent bedding</td><td>Harlan</td><td>7092</td><td></td></tr><tr><td>Nestlets</td><td>Ancare</td><td>-</td><td>Made of pulped virgin cotton fiber, dust-free and autoclavable</td></tr><tr><td>50 mL Conical tubes</td><td>Fisher Scientific</td><td>14-432-22</td><td>May use any brand, must be sterile</td></tr><tr><td>Isoflurane USP, 99.9 %, inhalation anesthetic</td><td>Pharmaceutical Partners of Canada Inc.</td><td>M60302</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td>Can prepare in lab</td></tr><tr><td>10 % Neutral Buffered Formalin</td><td>Sigma-Aldrich</td><td>HT501128</td><td>May use any brand</td></tr><tr><td>NAPCO NapFlow 1200 Class II A/B3 Biosafety Microbiological Safety Cabinet (cell culture hood)</td><td>NAPCO</td><td>Model used not currently available</td><td>May use any brand</td></tr><tr><td>Thermo Fisher Scientific Precision Heated Water Bath</td><td>Fisher Scientific</td><td>Model used not currently available&amp;nbsp;</td><td>May use any brand</td></tr><tr><td colspan="4">[header]</td></tr><tr><td>Reichert Bright-line Hemacytometer</td><td>Sigma-Aldrich</td><td>Z359629</td><td>May use any brand</td></tr><tr><td>Typhoon Trio BioAnalyzer&amp;nbsp;</td><td>GE Healthcare Life Sciences</td><td>Model used not currently available&amp;nbsp;</td><td>May use any fluorescence plate reader</td></tr><tr><td>Tecan Safire&lt;sup&gt;2&lt;/sup&gt; Multi-detection Microplate Reader</td><td>Tecan</td><td>Model used not currently available&amp;nbsp;</td><td>May use any fluorescence plate reader</td></tr><tr><td>Allegra 6R benchtop centrifuge</td><td>Beckman Coulter</td><td>366816</td><td>May use any brand</td></tr><tr><td>Table Top Anaesthesia machine</td><td>VetEquip</td><td>Model used not currently available&amp;nbsp;</td><td>May use any brand, must be portable</td></tr><tr><td>Wahl Peanut Mini Clippers</td><td>Wahl</td><td></td><td>May use any brand of small clippers</td></tr><tr><td>Insulin syringes 29 G x 1/2&#039;, 0.3 mL</td><td>BD</td><td>329464</td><td>May use any brand. Insulin syringes are recommended as they make injections easier through the rat&amp;rsquo;s tough skin.&amp;nbsp;</td></tr><tr><td>Cotton swabs</td><td>MedPro</td><td>018-425</td><td>May use any brand</td></tr><tr><td>Sharp-Pointed Dissecting Scissors</td><td>Fisher Scientific</td><td>8940</td><td>May use any brand</td></tr><tr><td>Dissecting Tissue Forceps</td><td>Fisher Scientific</td><td>13-812-41</td><td>May use any brand</td></tr></tbody></table>

handling of the cotton rat in studies for the pre-clinical evaluation of oncolytic viruses

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact. For this strategy to be successful, recent studies have shown that involvement of the host immune system is essential. Therefore, oncolytic virotherapy should be evaluated within the context of an immunocompetent model. Furthermore, the study of antitumor therapies in tolerized animal models may better recapitulate results seen in clinical trials. Cotton rats, commonly used to study respiratory viruses, are an attractive model to study oncolytic virotherapy as syngeneic models of mammary carcinoma and osteosarcoma are well established. However, there is a lack of published information on the proper handling procedure for these highly excitable rodents. The handling and capture approach outlined minimizes animal stress to facilitate experimentation. This technique hinges upon the ability of the researcher to keep calm during handling and perform procedures in a timely fashion. Finally, we describe how to prepare cotton rat mammary tumor cells for consistent subcutaneous tumor formation, and how to perform intratumoral and intraperitoneal injections. These methods can be applied to a wide range of studies furthering the development of the cotton rat as a relevant pre-clinical model to study antitumor therapy.

בשל האופי מאוד הרגיש של חולדות כותנה, היכרות עם הנהלים וניצול מותאמים כדי להפחית את הלחץ של בעלי החיים יקל בשימוש בם כמודל חיה פרה-קליני. שימוש בטכניקות טיפול הולמים גם למזער את הסיכון לחוקר. בעת שימוש בחולדות כותנה זה הכרחי כדי ל?...

Watch this Scientific Journal Video about Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses at JoVE.com

Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses

Cotton rats are extremely excitable and have a strong flight-or-fight response. A handling method optimized to reduce the stress of the animals is described which will make cotton rats more accessible as a preclinical model.

טיפול של הכותנה העכברוש במחקרים להערכה טרום-הקלינית של וירוסי Oncolytic

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact.  For this strategy to be ...

handling-cotton-rat-studies-for-pre-clinical-evaluation-oncolytic

Research

JoVE Journal

Medicine

9.3K Views.  McMaster University. Cotton rats are extremely excitable and have a strong flight-or-fight response. A handling method optimized to reduce the stress of the animals is described which will make cotton rats more accessible as a preclinical model. 

Video: Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses

Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy

High-throughput genome-wide RNAi (RNA interference) screening technology has been widely used for discovering host factors that impact&#160;virus replication. Here we present the application of this technology to uncovering host targets that specifically modulate the replication of Maraba virus, an oncolytic rhabdovirus, and vaccinia virus with the goal of enhancing therapy. While the protocol has been tested for use with oncolytic Maraba virus and oncolytic vaccinia virus, this approach is applicable to other oncolytic viruses and can also be utilized for identifying host targets that modulate virus replication in mammalian cells in general. This protocol describes the development and validation of an assay for high-throughput RNAi screening in mammalian cells, the key considerations and preparation steps important for conducting a primary high-throughput RNAi screen, and a step-by-step guide for conducting a primary high-throughput RNAi screen; in addition, it broadly outlines the methods for conducting secondary screen validation and tertiary validation studies. The benefit of high-throughput RNAi screening is that it allows one to catalogue, in an extensive and unbiased fashion, host factors that modulate any aspect of virus replication for which one can develop an in vitro assay such as infectivity, burst size, and cytotoxicity. It has the power to uncover biotherapeutic targets unforeseen based on current knowledge.

Here we describe a protocol for employing high-throughput RNAi screening to uncover host targets that can be manipulated to enhance oncolytic virus therapy, specifically rhabodvirus and vaccinia virus therapy, but it can be readily adapted to other oncolytic virus platforms or for discovering host genes that modulate virus replication generally.

ברמת הגנום RNAi ההקרנה כדי לזהות גורמים מארח לווסת את הטיפול Oncolytic וירוס

High-throughput genome-wide RNAi (RNA interference) screening technology has been widely used for discovering host factors that impact&#160;virus ...

Cancer Research

A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different forms of learning. Specifically, viral-mediated delivery is used to express a genetically modified inhibitory G-protein coupled receptor (the Designer Receptor) into a discrete brain region in the rodent. Three weeks later, when designer receptor expression levels are high, a pharmacological agent (the Designer Drug) is administered systemically 30 min prior to a specific behavioral session. The drug has affinity for the designer receptor and thus results in inhibition of neurons that express the designer receptor, but is otherwise biologically inert. The brain region remains silenced for 2-5 hr (depending on the dose and route of administration). Upon completion of the behavioral paradigm, brain tissue is assessed for correct placement and receptor expression. This approach is particularly useful for determining the contribution of individual brain regions to specific components of behavior and can be used across any number of behavioral paradigms.

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

שיטה להשתקה מרחוק עצבית פעילות במכרסמים בשלבים בדידים של למידה

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and ...

Behavior

Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis. In recent years, oncolytic virotherapies have proven themselves to be potentially powerful tools to fight malignancy. Due to the unique dual blood supply in the liver, it is possible to apply therapies locally to orthotopic liver tumors, which are predominantly fed by arterial blood flow. We have previously demonstrated that hepatic arterial delivery of oncolytic viruses results in safe and efficient transduction efficiency of multifocal HCC lesions, resulting in significant prolongation of survival in immune competent rats. This procedure closely mimics the application of transarterial embolization in patients, which is the standard palliative care provided to many HCC patients. The ability to administer tumor therapies through the hepatic artery in rats allows for a highly sophisticated preclinical model for evaluating novel viral vectors under development. Here we describe the detailed protocol for microdissection of the hepatic artery for infusion of oncolytic virus vectors to treat orthotopic HCC.

Oncolytic virotherapies are under development as novel therapeutics for the treatment of hepatocellular carcinoma (HCC). Here we describe a method for locoregional therapy of HCC via hepatic arterial administration of oncolytic virus.

Transarterial מינהל oncolytic וירוסים לטיפול Locoregional של Orthotopic HCC אצל חולדות

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis.  In recent years, oncolytic virotherapies have proven ...

Generation of Transgenic Rats using a Lentiviral Vector Approach

Transgenic animal models are fundamentally important for modern biomedical research. The incorporation of foreign genes into early mouse or rat embryos is an invaluable tool for gene function analysis in living organisms. The standard transgenesis method is based on microinjecting foreign DNA fragments into a pronucleus of a fertilized oocyte. This technique is widely used in mice but remains relatively inefficient and technically demanding in other animal species. The transgene can also be introduced into one-cell-stage embryos via lentiviral infection, providing an effective alternative to standard pronuclear injections, especially in species or strains with a more challenging embryo structure. In this approach, a suspension that contains lentiviral vectors is injected into the perivitelline space of a fertilized rat embryo, which is technically less demanding and has a higher success rate. Lentiviral vectors were shown to efficiently incorporate the transgene into the genome to determine the generation of stable transgenic lines. Despite some limitations (e.g., Biosafety Level 2 requirements, DNA fragment size limits), lentiviral transgenesis is a rapid and efficient transgenesis method. Additionally, using female rats that are mated with a fertile male strain with a different dominant fur color is presented as an alternative to generate pseudopregnant foster mothers.

This article aims to provide the methodology for lentiviral transgenesis in rat embryos using multiple injections of a virus suspension into the zygote perivitelline space. Female rats that are mated with a fertile male strain with a different dominant fur color is used to generate pseudopregnant foster mothers.

הדור של חולדות הטרנסגניים באמצעות גישה וקטורית לנטינגיי

Transgenic animal models are fundamentally important for modern biomedical research. The incorporation of foreign genes into early mouse or rat embryos is ...

Bioengineering

Methods and Tips for Intravenous Administration of Adeno-associated Virus to Rats and Evaluation of Central Nervous System Transduction

Adeno-associated virus (AAV) vectors are a key reagent in the neurosciences for clustered regularly interspaced short palindromic repeats (CRISPR), optogenetics, cre-lox targeting, etc. The purpose of this manuscript is to aid the investigator attempting expansive central nervous system (CNS) gene transfer in the rat via tail vein injection of AAV. Wide-scale expression is relevant for conditions with widespread pathology, and a rat model is significant due to its greater size and physiologic similarities to humans compared to mice. In this example application, a wide-scale neuronal transduction is used to mimic a neurodegenerative disease that affects the entire spinal cord, amyotrophic lateral sclerosis (ALS). The efficient wide-scale CNS transduction can also be used to deliver therapeutic protein factors in pre-clinical studies. After a post-injection expression interval of several weeks, the effects of the transduction are evaluated. For a green fluorescent protein (GFP) control vector, the amount of GFP in the cerebellum is estimated quickly and reliably by a basic imaging program. For motor disease phenotypes that are induced by the ALS related protein transactive response DNA-binding protein of 43 kDa (TDP-43), the deficits are scored by escape reflex and rotarod. Beyond disease modeling and gene therapy, there are diverse potential applications for the wide-scale gene targeting described here. The expanded use of this method will aid in expediting hypothesis testing in the neurosciences and neurogenetics.

Methods for a wide-scale central nervous system gene delivery in the rat are covered. In this example, the purpose is to mimic a disease that affects the entire spinal cord. The widespread transduction can be used to deliver a therapeutic protein to the CNS from a one-time, peripheral administration.

שיטות וטיפים על עירוי לוריד של Adeno-הקשורים וירוס חולדות והערכה של מערכת העצבים המרכזית התמרה חושית

Adeno-associated virus (AAV) vectors are a key reagent in the neurosciences for clustered regularly interspaced short palindromic repeats (CRISPR), ...

Neuroscience

Humanized NOG Mice for Intravaginal HIV Exposure and Treatment of HIV Infection

Humanized mice provide a sophisticated platform to study human immunodeficiency virus (HIV) virology and to test antiviral drugs. This protocol describes the establishment of a human immune system in adult NOG mice. Here, we explain all the practical steps from isolation of umbilical cord blood derived human CD34+ cells and their subsequent intravenous transplantation into the mice, to the manipulation of the model through HIV infection, combination antiretroviral therapy (cART), and blood sampling. Approximately 75,000 hCD34+ cells are injected intravenously into the mice and the level of human chimerism, also known as humanization, in the peripheral blood is estimated longitudinally for months by flow cytometry. A total of 75,000 hCD34+ cells yields 20%&#8211;50% human CD45+ cells in the peripheral blood. The mice are susceptible to intravaginal infection with HIV and blood can be sampled once weekly for analysis, and twice monthly for extended periods. This protocol describes an assay for quantification of plasma viral load using droplet digital PCR (ddPCR). We show how the mice can be effectively treated with a standard-of-care cART regimen in the diet. The delivery of cART in the form of regular mouse chow is a significant refinement of the experimental model. This model can be used for preclinical analysis of both systemic and topical pre-exposure prophylaxis compounds as well as for testing of novel treatments and HIV cure strategies.

We have developed a protocol for the generation and evaluation of a humanized and human immunodeficiency virus-infected NOG mouse model based on stem cell transplant, intravaginal human immunodeficiency virus exposure, and droplet digital PCR RNA quantification.

הומניזציה חעוגה עכברים עבור חשיפה Intravaginal HIV וטיפול בזיהום HIV

Humanized mice provide a sophisticated platform to study human immunodeficiency virus (HIV) virology and to test antiviral drugs. This protocol describes ...

Immunology and Infection

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

מודל עכברוש של זיהום מוחי EcoHIV

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...

Viral Transgene Expression in Rodent Hearts and the Assessment of Cardiac Arrhythmia Risk

Heart disease is the leading cause of morbidity and mortality worldwide. Due to the ease of handling&#160;and abundance of transgenic strains, rodents have become essential models for cardiovascular research. However, spontaneous lethal cardiac arrhythmias that often cause mortality in heart disease patients are rare in rodent models of heart disease. This is primarily due to the species differences in cardiac electrical properties between human&#160;and rodents and poses a challenge to the study of cardiac arrhythmias using rodents. This protocol describes an approach to enable efficient transgene expression in mouse and rat ventricular myocardium using echocardiography-guided intramuscular injections of recombinant virus (adenovirus and adeno-associated virus). This work also outlines a method to enable reliable assessment of cardiac susceptibility to arrhythmias using isolated, Langendorff-perfused mouse and rat hearts with both adrenergic and programmed electrical stimulations. These techniques are critical for studying heart rhythm disorders associated with adverse cardiac remodeling after injuries, such as myocardial infarction.

The present protocol describes methods for transgene expression in rat and mouse hearts by direct intramyocardial injection of the virus under echocardiography guidance. Methods for the assessment of the susceptibility of hearts to ventricular arrhythmias by the programmed electrical stimulation of isolated, Langendorff-perfused hearts are also explained here.

ביטוי טרנסגני נגיפי בלבבות מכרסמים והערכת הסיכון להפרעות קצב לב

Heart disease is the leading cause of morbidity and mortality worldwide. Due to the ease of handling&#160;and abundance of transgenic strains, rodents ...

Biology

Establishing an Active HIV Infection in a Rat Model

Source: Li, H., McLaurin, et al. <a href="https://app.jove.com/v/62137">A Rat Model of EcoHIV Brain Infection.</a> J. Vis. Exp. (2021).
This video demonstrates the procedure of injecting chimeric HIV viruses into the cortex of a rat&#39;s brain, where the virus RNA integrates into the host genome, replicates, and spreads to establish an active HIV infection in the experimental model.

הקמת זיהום HIV פעיל במודל חולדה

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
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JoVE Encyclopedia of Experiments

Neuropathology

Infectious Diseases

Targeting Liver Tumors with Oncolytic Viruses via the Hepatic Artery

Source: Altomonte, J. et al., <a href="https://app.jove.com/v/53757">Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats.</a>&#160;J. Vis. Exp. (2016)
The video demonstrates locoregional delivery of oncolytic viruses through hepatic arterial injection as a therapeutic approach for hepatocellular carcinoma. These viruses selectively replicate in tumor cells, inducing cell lysis and releasing tumor cell-associated antigens. This process triggers immunogenic cell death, inducing an anti-tumor response.