scispace - formally typeset
Search or ask a question
Journal ArticleDOI

Retro-orbital injections in mice.

01 May 2011-Lab Animal (NIH Public Access)-Vol. 40, Iss: 5, pp 155-160
TL;DR: The authors report that they have routinely used retro-orbital injection of the venous sinus in the adult mouse to administer volumes up to 150 μl without incident and this technique is a useful alternative to tail vein injection for the administration of non-tumorigenic compounds.
Abstract: Intravenous vascular access is technically challenging in the adult mouse and even more challenging in neonatal mice. The authors describe the technique of retro-orbital injection of the venous sinus in the adult and neonatal mouse. This technique is a useful alternative to tail vein injection for the administration of non-tumorigenic compounds. The authors report that they have routinely used this technique in the adult mouse to administer volumes up to 150 μl without incident. Administration of retro-orbital injections is more challenging in neonatal mice but can reliably deliver volumes up to 10 μl.

Content maybe subject to copyright    Report

Citations
More filters
Journal ArticleDOI
16 Aug 2019-Science
TL;DR: A GEVI is engineered, called Voltron, that uses bright and photostable synthetic dyes instead of protein-based fluorophores, thereby extending the number of neurons imaged simultaneously in vivo by a factor of 10 and enabling imaging for significantly longer durations relative to existing GEVIs.
Abstract: Genetically encoded voltage indicators (GEVIs) enable monitoring of neuronal activity at high spatial and temporal resolution. However, the utility of existing GEVIs has been limited by the brightness and photostability of fluorescent proteins and rhodopsins. We engineered a GEVI, called Voltron, that uses bright and photostable synthetic dyes instead of protein-based fluorophores, thereby extending the number of neurons imaged simultaneously in vivo by a factor of 10 and enabling imaging for significantly longer durations relative to existing GEVIs. We used Voltron for in vivo voltage imaging in mice, zebrafish, and fruit flies. In the mouse cortex, Voltron allowed single-trial recording of spikes and subthreshold voltage signals from dozens of neurons simultaneously over a 15-minute period of continuous imaging. In larval zebrafish, Voltron enabled the precise correlation of spike timing with behavior.

313 citations

Journal ArticleDOI
TL;DR: A novel pathophysiological role is established for CMNLRP3 inflammasome signaling, with a mechanistic link to the pathogenesis of AF, and the inhibition of NLRP3 is established as a potential novel AF therapy approach.
Abstract: Background —Atrial fibrillation (AF) is frequently associated with enhanced inflammatory response. The "NACHT, LRR and PYD domain containing protein 3" (NLRP3)-inflammasome mediates caspase-1 activation and interleukin-1β release in immune cells, but is not known to play a role in cardiomyocytes (CMs). Here, we assessed the role of CM NLRP3-inflammasome in AF. Methods —NLRP3-inflammasome activation was assessed by immunoblot in atrial whole-tissue lysates and CMs from patients with paroxysmal (pAF) or long-standing persistent (chronic) AF (cAF). To determine whether CM-specific activation of NLPR3 is sufficient to promote AF, a CM-specific knock-in mouse model expressing constitutively active NLRP3 (CM-KI) was established. In vivo electrophysiology was used to assess atrial arrhythmia vulnerability. To evaluate the mechanism of AF, electrical activation pattern, Ca 2+ spark frequency (CaSF), atrial effective refractory period (AERP), and morphology of atria were evaluated in CM-KI mice and WT littermates. Results —NLRP3-inflammasome activity was increased in atrial CMs of pAF and cAF patients. CM-KI mice developed spontaneous premature atrial contractions and inducible AF, which was attenuated by a specific NLRP3-inflammasome inhibitor, MCC950. CM-KI mice exhibited ectopic activity, abnormal sarcoplasmic-reticulum Ca 2+ -release, AERP shortening and atrial hypertrophy. Adeno-associated virus subtype-9 mediated CM-specific knockdown of Nlrp3 suppressed AF development in CM-KI mice. Finally, genetic inhibition of Nlrp3 prevented AF development in CREM transgenic mice, a well-characterized mouse model of spontaneous AF. Conclusions —Our study establishes a novel pathophysiological role for CM NLRP3-inflammasome signaling with a mechanistic link to the pathogenesis of AF, and establishes inhibition of NLRP3 as a potential novel AF-therapy approach.

305 citations

Journal ArticleDOI
TL;DR: Having developed AAV capsids that target sites throughout the body, the authors describe how to produce and systemically administer these AAVs to rodents to label and/or genetically manipulate cells in the nervous system and visceral organs.
Abstract: We recently developed adeno-associated virus (AAV) capsids to facilitate efficient and noninvasive gene transfer to the central and peripheral nervous systems. However, a detailed protocol for generating and systemically delivering novel AAV variants was not previously available. In this protocol, we describe how to produce and intravenously administer AAVs to adult mice to specifically label and/or genetically manipulate cells in the nervous system and organs, including the heart. The procedure comprises three separate stages: AAV production, intravenous delivery, and evaluation of transgene expression. The protocol spans 8 d, excluding the time required to assess gene expression, and can be readily adopted by researchers with basic molecular biology, cell culture, and animal work experience. We provide guidelines for experimental design and choice of the capsid, cargo, and viral dose appropriate for the experimental aims. The procedures outlined here are adaptable to diverse biomedical applications, from anatomical and functional mapping to gene expression, silencing, and editing.

200 citations

Journal ArticleDOI
TL;DR: Results showed that mitoTALENs, when expressed in affected tissues, could revert disease-related phenotypes in mice and reduce mutation burden and correct biochemical defects in a mouse model of mitochondrial disease.
Abstract: Mutations in the mitochondrial DNA (mtDNA) are responsible for several metabolic disorders, commonly involving muscle and the central nervous system1 Because of the critical role of mtDNA in oxidative phosphorylation, the majority of pathogenic mtDNA mutations are heteroplasmic, co-existing with wild-type molecules1 Using a mouse model with a heteroplasmic mtDNA mutation2, we tested whether mitochondrial-targeted TALENs (mitoTALENs)3,4 could reduce the mutant mtDNA load in muscle and heart AAV9-mitoTALEN was administered via intramuscular, intravenous, and intraperitoneal injections Muscle and heart were efficiently transduced and showed a robust reduction in mutant mtDNA, which was stable over time The molecular defect, namely a decrease in transfer RNAAla levels, was restored by the treatment These results showed that mitoTALENs, when expressed in affected tissues, could revert disease-related phenotypes in mice

171 citations

Journal ArticleDOI
05 Nov 2019-Mbio
TL;DR: It is demonstrated that epimutation can both be induced and reverted after in vivo passage through a mouse; rates of both induction and reversion are higher after brain infection than after infection of other organs (liver, spleen, kidneys, or lungs).
Abstract: The environmentally ubiquitous fungus Mucor circinelloides is a primary cause of the emerging disease mucormycosis. Mucor infection is notable for causing high morbidity and mortality, especially in immunosuppressed patients, while being inherently resistant to the majority of clinically available antifungal drugs. A new, RNA interference (RNAi)-dependent, and reversible epigenetic mechanism of antifungal resistance-epimutation-was recently discovered in M. circinelloides However, the effects of epimutation in a host-pathogen setting were unknown. We employed a systemic, intravenous murine model of Mucor infection to elucidate the potential impact of epimutation in vivo Infection with an epimutant strain resistant to the antifungal agents FK506 and rapamycin revealed that the epimutant-induced drug resistance was stable in vivo in a variety of different organs and tissues. Reversion of the epimutant-induced drug resistance was observed to be more rapid in isolates from the brain than in isolates recovered from the liver, spleen, kidney, or lungs. Importantly, infection with a wild-type strain of Mucor led to increased rates of epimutation after strains were recovered from organs and exposed to FK506 stress in vitro. Once again, this effect was more pronounced in strains recovered from the brain than from other organs. In summary, we report the rapid induction and reversion of RNAi-dependent drug resistance after in vivo passage through a murine model, with pronounced impact in strains recovered from brain. Defining the role played by epimutation in drug resistance and infection advances our understanding of Mucor and other fungal pathogens and may have implications for antifungal therapy.IMPORTANCE The emerging fungal pathogen Mucor circinelloides causes a severe infection, mucormycosis, which leads to considerable morbidity and mortality. Treatment of Mucor infection is challenging because Mucor is inherently resistant to nearly all clinical antifungal agents. An RNAi-dependent and reversible mechanism of antifungal resistance, epimutation, was recently reported for Mucor Epimutation has not been studied in vivo, and it was unclear whether it would contribute to antifungal resistance observed clinically. We demonstrate that epimutation can both be induced and reverted after in vivo passage through a mouse; rates of both induction and reversion are higher after brain infection than after infection of other organs (liver, spleen, kidneys, or lungs). Elucidating the roles played by epimutation in drug resistance and infection will improve our understanding of Mucor and other fungal pathogens and may have implications for antifungal treatment.

135 citations

References
More filters
Book
01 Feb 1996

13,908 citations

01 Jan 2007
TL;DR: The Mouse in biomedical research , The Mouse inomedical research , مرکز فناوری اطلاعات و £1,000,000; اوشاوρزی; کسورز رسانی ;
Abstract: The Mouse in biomedical research , The Mouse in biomedical research , مرکز فناوری اطلاعات و اطلاع رسانی کشاورزی

536 citations

Journal ArticleDOI
TL;DR: It is found that Mst1 and Mst2, the two mouse homologs of the Drosophila Hpo, control the sizes of some, but not all organs, in mice, and MSt1 andMst2 act as tumor suppressors by restricting cell proliferation and survival.
Abstract: Control of organ size by cell proliferation and survival is a fundamental developmental process, and its deregulation leads to cancer. However, the molecular mechanism underlying organ size control remains elusive in vertebrates. In Drosophila, the Hippo (Hpo) signaling pathway controls organ size by both restricting cell growth and proliferation and promoting cell death. Here we investigated whether mammals also require the Hpo pathway to control organ size and adult tissue homeostasis. We found that Mst1 and Mst2, the two mouse homologs of the Drosophila Hpo, control the sizes of some, but not all organs, in mice, and Mst1 and Mst2 act as tumor suppressors by restricting cell proliferation and survival. We show that Mst1 and Mst2 play redundant roles, and removal of both resulted in early lethality in mouse embryos. Importantly, tumors developed in the liver with a substantial increase of the stem/progenitor cells by 6 months after removing Mst1 and Mst2 postnatally. We show that Mst1 and Mst2 were required in vivo to control Yap phosphorylation and activity. Interestingly, apoptosis induced by TNFα was blocked in the Mst1 and Mst2 double-mutant cells both in vivo and in vitro. As TNFα is a pleiotropic inflammatory cytokine affecting most organs by regulating cell proliferation and cell death, resistance to TNFα-induced cell death may also contribute significantly to tumor formation in the absence of Mst1 and Mst2.

508 citations

Journal ArticleDOI
26 Feb 2010-Immunity
TL;DR: The results reveal insight into the dynamic regulation of T cell:B cell interactions and identify SLAM family members as critical components of sustained T cell-B cell adhesion required for productive humoral immunity.

346 citations

Book
27 Dec 2000
TL;DR: This chapter discusses the development of Phenotyping Terminology and Tests, and some of the techniques used to develop and evaluate these techniques.
Abstract: Important Biological Features Introduction Stocks and Strains Nomenclature Behavior Anatomic and Physiologic Features Normative Values Hematology Immunodeficient Mice Wild Mice References Husbandry Housing Barriers and Containment Personal Protective Equipment (PPE) Special Considerations for Immunodeficient Mice Environment Sanitation and Pest Control Nutrition Water Breeding Identification and Record Keeping Transportation References Management Regulatory Agencies and Compliance Institutional Animal Care and Use Committee (IACUC) Occupational Health and Zoonotic Diseases References Clinical Medicine Basic Veterinary Supplies Physical Examination of the Mouse Common Spontaneous and Noninfectious Diseases Common Infectious Diseases Treatment and Supportive Care of Sick Mice Clinical Endpoints Treating Disease on a Colony Basis References Preventive Medicine Receiving Testing of Biological Materials Health Surveillance and Monitoring Disease Prevention through Sanitation References Experimental Methodology Restraint Sampling Methods Compound Administration Phenotyping Anesthesia and Analgesia Euthanasia Necropsy References Resources and Additional Information Organizations Publications Electronic Resources Appendix A: Glossary of Phenotyping Terminology and Tests Appendix B: Suggested Cassette Numbering System and Some Trimming Suggestions Index

316 citations