JoVE Logo

Sign In

National Institute of Environmental Health Sciences

10 ARTICLES PUBLISHED IN JoVE

image

Biology

Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal
Xiaofeng Zheng 1, Guang Hu 1
1Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences

We describe a fluorescence reporter assay to quickly identify and characterize genes that regulate mouse embryonic stem cell maintenance and self-renewal.

image

Developmental Biology

Quantifying Branching Density in Rat Mammary Gland Whole-mounts Using the Sholl Analysis Method
Jason P. Stanko 1, Suzanne E. Fenton 1
1National Toxicology Program Laboratory, Division of the National Toxicology Program, National Institute of Environmental Health Sciences

Mammary gland development in the rodent has typically been evaluated using descriptive assessments or by measuring basic physical attributes. Branching density is an indicator of mammary development that is difficult to quantify objectively. This protocol describes a reliable method for the quantitative assessment of mammary gland branching characteristics.

image

Cancer Research

Sectioning Mammary Gland Whole Mounts for Lesion Identification
Deirdre K Tucker 1,2, Julie F Foley 3, Schantel A Bouknight 4, Suzanne E Fenton 2
1Curriculum in Toxicology, University of North Carolina at Chapel Hill, 2National Toxicology Program Laboratory (NTPL), DNTP, National Institute of Environmental Health Sciences, 3Cellular and Molecular Pathology Branch, DNTP, National Institute of Environmental Health Sciences, 4Charles River Laboratories Inc.

We developed a method to successfully remove, process, section, and stain, for histopathological evaluation, mammary tissue that had originally been fixed on slides as whole mounts. This method may promote the collection and evaluation of mammary gland whole mounts in reproductive and developmental test guideline studies.

image

Developmental Biology

A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse
Mary C. Peavey 1,4, Corey L. Reynolds 2, Maria M. Szwarc 2, William E. Gibbons 1, Cecilia T. Valdes 1, Francesco J. DeMayo *3, John P. Lydon *4
1Devision of Repreoductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Baylor College of Medicine, 2Mouse Phenotyping Core, Baylor College of Medicine, 3Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, 4Department of Molecular and Cellular Biology, Baylor College of Medicine

Mice are widely used to study gestational biology. However, pregnancy termination is required for such studies which precludes longitudinal investigations and necessitates the use of large numbers of animals. Therefore, we describe a non-invasive technique of high-frequency ultrasonography for early detection and monitoring of post-implantation events in the pregnant mouse.

image

Biology

Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates
Gregory J. Scott 1, Artiom Gruzdev 1, Thomas B. Hagler 1, Manas K. Ray 1
1Knock Out Mouse Core, NIEHS/NIH

Here we present a protocol to increase chimera production without the use of new equipment. A simple orientation change of the embryo for injection can increase the number of embryos produced, and potentially reduce the timeline to germline transmission.

image

Biology

Co-immunoprecipitation Assay Using Endogenous Nuclear Proteins from Cells Cultured Under Hypoxic Conditions
Xiaofeng Zheng 1,2, Calvin Qing Wei Ho 1, Xiaowei Zheng 3, Kian Leong Lee 4, Katarina Gradin 5, Teresa S. Pereira 3, Per-Olof Berggren 1,2,3, Yusuf Ali 1,2
1Lee Kong Chian School of Medicine, Nanyang Technological University, 2Singapore Eye Research Institute (SERI), Singapore General Hospital, 3The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, 4Cancer and Stem Cell Biology Program, Duke-NUS Medical School, 5Department of Cell and Molecular Biology, Karolinska Institutet

Here we describe a co-immunoprecipitation protocol to study protein-protein interactions between endogenous nuclear proteins under hypoxic conditions. This method is suitable for demonstration of the interactions between transcription factors and transcriptional co-regulators at hypoxia.

image

JoVE Journal

Laser-assisted Lentiviral Gene Delivery to Mouse Fertilized Eggs
Negin P. Martin *1, Page Myers *2, Eugenia Goulding 1, Shih-Heng Chen 1, Mitzie Walker 1, Thomas M. Porter 1, Lucas Van Gorder 1, Amanda Mathew 1, Artiom Gruzdev 3, Erica Scappini 4, Charles Romeo 1
1Neurobiology Laboratory, National Institute of Environmental Health Sciences, 2Comparative Medicine Branch, National Institute of Environmental Health Sciences, 3Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, 4Signal Transduction Laboratory, National Institute of Environmental Health Sciences

Mouse fertilized eggs and early stage embryos are protected by the zona pellucida, a glycoprotein matrix that forms a barrier against gene delivery. This article describes a protocol for perforating the zona with a laser to transduce embryonic cells with lentiviral vectors and to create transgenic mice.

image

Genetics

Use of Frozen Tissue in the Comet Assay for the Evaluation of DNA Damage
Cheryl A. Hobbs *1, Leslie Recio 1, John Winters 1, Kristine L. Witt 2
1Toxicology Program, ILS, Inc., 2Division of the National Toxicology Program, National Institute of Environmental Health Sciences

This protocol describes several procedures for preparing high quality frozen tissue samples at the time of necropsy for use in the comet assay to assess DNA damage: 1) minced tissue, 2) scraped epithelial cells from the gastrointestinal tract, and 3) cubed tissue samples, requiring homogenization using a tissue mincing device.

image

Biochemistry

Removal and Replacement of Endogenous Ligands from Lipid-Bound Proteins and Allergens
Alexander C. Y. Foo 1, Peter M. Thompson 1, Geoffrey A. Mueller 1
1Nuclear Magnetic Resonance Group, National Institute of Environmental Health Sciences

This protocol describes the removal of endogenous lipids from allergens, and their replacement with user-specified ligands through reverse-phase HPLC coupled with thermal annealing. 31P-NMR and circular dichroism allow for the rapid confirmation of ligand removal/loading, and the recovery of native allergen structure.

image

Behavior

Behavioral Training Procedures for Head-fixed Virtual Reality in Mice
Jared Karl Glorius 1, Leslie Wilson 1, Ayland Letsinger 2, Parth Shirolkar 1,3, Jesse Cushman 1
1Neurobehavioral Core, National Institute of Environmental Health Sciences, 2Ion Channel Physiology Group, National Institute of Environmental Health Sciences, 3University of North Carolina at Chapel Hill

The article describes the experimental procedures for the commonly used linear track virtual reality (VR) paradigm in mice as well as determining the feasibility of running complex VR tasks by testing a Y-shaped signal discrimination task.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved