Technologies

Research Tools : Animal models

Technologies

Invertebrate Models for Studying Traumatic Brain Injury

UW–Madison researchers have developed a method for inflicting closed head TBI in invertebrates. The method can be used to screen candidate therapeutic agents for treating TBI.

Specifically, the researchers created a device that mechanically induces TBI in organisms such as Drosophila melanogaster (fruit flies), which respond to impact trauma in many of the same ways as humans. Subjecting these organisms to a controlled impact and dosing with test agents could help identify the biological pathways associated with the consequences of TBI as well as enable screens for therapeutic compounds.

The device consists of a spring attached to a vial containing up to 100 flies. The spring is pulled back and then released, striking the vial against a rubber pad to inflict brain trauma as the flies collide with the bottom and walls of the vial. The device can be adjusted to inflict injury at differing severities.
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Mutation in the Rat Adenomatous Polyposis Coli Gene within the Human Mutation Hotspot Region

UW-Madison researchers have developed the Pirc rat, the first and only currently available knockout rat that develops easily visible tumors arising from mutations in the APC gene. They used N-ethyl-N-nitrosourea mutagenesis to create a truncating mutation in the rat APC gene at a site corresponding to the mutation hotspot region of the human ortholog. This germline mutation was confirmed by sequencing and has been transmitted to approximately half the offspring of the founder male.

Unlike existing rodent models, these rats develop multiple adenomatous polyps in the colon, more closely imitating human colorectal cancer. They may be used to screen potential carcinogens or compounds that could prevent carcinogenesis.
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Assay to Detect Viral Uncoating

UW-Madison researchers have developed a cell-free system for studying the viral uncoating mechanism. This system uses a sub group of avian sarcoma and leucosis virus (ASLV-A) as a model retrovirus. The TVA receptor for ASLV-A is incorporated into the human cell line 293. After ASLV-A enters the 293 cells and accumulates in the cell’s endosomes, the cells are lysed. The virus-containing endosomes are isolated under conditions of high pH, which block entry of the virus into the cytosol, providing a cell-free system for studying uncoating. Conditions can then be altered to determine the effect of various agents on viral uncoating and other post viral envelope-endosome fusion events. Experiments have shown that the viral nucleic acid released from the endosomes is competent for reverse transcription, and that viral DNA synthesis is dependent on cellular factors contained in the S100 or S10 fraction of the 293 cell line.
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Endothelial Cells Derived from Human Embryonic Stem Cells

UW-Madison researchers have developed a simple and efficient method of inducing human embryonic stem cells to differentiate into a relatively homogenous population of endothelial cells. The method involves culturing ES cells in a commercially available medium that supports the growth of endothelial cells. The resulting ES-derived endothelial cells have the general morphological characteristics and cell surface markers of endothelial cells. They are capable of inducing and participating in blood vessel formation when transplanted into tissue in vivo.
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Transgenic Mouse Expressing Green Fluorescent Protein in Glial Cells

UW-Madison researchers have developed a transgenic mouse containing a fluorescent reporter gene linked to the glial fibrillary acidic protein promoter. The mouse was engineered by inserting a genetic construct containing a GFAP promoter, a humanized, mutant green fluorescent reporter (GFP) gene and a polyadenylation signal sequence, into the pronucleus of a zygote.

The mouse can be used in a non-invasive assay of neurotoxicity. In the assay, a mouse is exposed to a suspected neurotoxic agent. The presence of green fluorescence signal in glial cells, such as astrocytes, Mueller cells or Schwann cells, indicates the GFAP promoter has been up-regulated in response to chemical or physical injury to cells, confirming the neurotoxicity of the substance.
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Laboratory Mouse Procedural Techniques Training DVD and Manual

Trainers at the RARC have developed a training DVD that can be used as a resource or refresher for procedural techniques on mice. This DVD, called “Laboratory Mouse – Procedural Techniques,” provides content similar to that of the in-person training course offered through the RARC.

Procedures and techniques demonstrated in the DVD include handling, restraint, oral gavage, identification and common bleeding and injection techniques. All procedures and techniques shown in the DVD are approved by the University of Wisconsin’s Animal Care and Use Committee (ACUC).
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Laboratory Rat Procedural Techniques Training DVD and Manual

Trainers at the RARC have developed a training DVD that can be used as a resource or refresher for procedural techniques on rats. This DVD, called “Laboratory Rat – Procedural Techniques,” provides content similar to that of the in-person training course offered through the RARC.

Procedures and techniques demonstrated in the DVD include handling, restraint, oral gavage, identification and common bleeding and injection techniques. All procedures and techniques shown in the DVD are approved by the University of Wisconsin’s Animal Care and Use Committee (ACUC).
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Transgenic Mouse for Testing Chemotherapeutic Agents

UW-Madison researchers have developed transgenic mice that are conditional knockouts for mdm2. In these mice, exons 7-9 of the mdm2 gene are flanked with a particular sequence. When exposed to a specific recombinase, these exons should be deleted, rendering the gene non-functional.
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Transgenic Mice Containing an Antioxidant Response Element-Driven Reporter Gene

UW-Madison researchers have developed a line of transgenic mice that contain a 51 base pair region of human ARE paired with the human placental alkaline phosphatase (hPAP) gene as a reporter. The researchers chose hPAP as a reporter gene because it is insensitive to heat and its activity is relatively easy to quantify, both of which should facilitate high throughput screening.
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Mouse Expressed Sequence Tags Related to Liver, Skin, Kidney, Thymus, Lung and Palate Toxicology

UW-Madison researchers have developed tools for monitoring changes in gene expression associated with exposure to various tissue-specific toxicants in the mouse model. Their approach involves determining the frequency of expressed sequence tags (ESTs), which are 200 to 500 base pair sequences of genomic DNA generated from cDNA.

The researchers first constructed cDNA libraries from mRNA extracted from various tissues of control mice and the livers of mice exposed to certain toxicants; sequenced the ends of the cDNA clones to create ESTs; and then organized the clones and counted them. Comparing EST libraries of control and toxin-exposed livers allows researchers to determine how toxicant exposure affects gene expression.

The EST frequency approach also generates a reagent for use in cDNA microarrays, allowing more rapid comparison of expression patterns between samples. Thus, this technology includes three reagents: EST clone sets from various mouse tissues, an organized version of this information and arrays of cDNAs in either liquid culture or as spots in microarrays.
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Transgenic Rats Expressing Human Placental Alkaline Phosphatase

UW-Madison researchers have now developed transgenic rats that ubiquitously express the marker gene human placental alkaline phosphatase (hPAP). They used an 800-base pair fragment of a promoter region from ROSA26 cells in a genetic construct with hPAP.

Several lines of transgenic rats were created using the microinjection technique. The ROSA26 promoter directs ubiquitous expression of hPAP during embryonic and postnatal development in the rats.
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Transgenic Mice Expressing Human Alkaline Phosphatase

UW-Madison researchers have now developed transgenic mice that ubiquitously express the marker gene human placental alkaline phosphatase (hPAP). They used an 800-base pair fragment of a promoter region from ROSA26 cells in a genetic construct with hPAP.

Several lines of transgenic mice from both the FVB/N and C57BL/6 mouse strains were created using the microinjection technique. The ROSA26 promoter directs ubiquitous expression of hPAP during embryonic and postnatal development in the mice.
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Transgenic Mice That Ubiquitously Express Enhanced Green Fluorescent Protein

UW-Madison researchers have developed transgenic mice that ubiquitously express the marker gene enhanced green fluorescent protein (EGFP). They used an 800-base pair fragment of a promoter region from ROSA26 cells in a genetic construct with EGFP.

Several lines of transgenic mice were created using the microinjection technique. The ROSA26 promoter directs ubiquitous expression of EGFP during embryonic and postnatal development in the mice.
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A Transgenic Mouse Model for Human Liver Disease

UW–Madison researchers have developed an excellent transgenic mouse model for human liver disease. They used a genetic construct comprising the albumin gene regulatory element linked to the herpes simplex thymidine kinase gene to create the mouse. When an inducing agent, such as the drug gancyclovir, is administered to the mouse, it develops liver disease.
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Targeted Disruption of the Murine NADPH Cytochrome P-450 Oxidoreductase Gene

The UW-Madison researchers have created a mouse line in which one or both alleles of the CYPOR gene have been disrupted. The knockout of both alleles is lethal, while heterozygotes show only 60 to 75 percent of the enzyme activity of controls and therefore display increased sensitivity to factors affecting cytochrome P450- and other CYPOR-dependent metabolic pathways. These animal models can be used to directly assess the potential risks of new, xenobiotic compounds. 
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Collection of Drosophila Mutants with Neurodegenerative Effects

UW-Madison researchers have compiled a collection of mutant Drosophila with obvious and distinct neurodegeneration.  Each of these mutants has a defect in a single gene.  They could be used to identify novel drug targets for neurodegenerative diseases.
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Expressible cDNA Encoding Human Calcium-Activated Potassium Channels

UW-Madison researchers have isolated cDNAs encoding the human and mouse homologs of the Drosophila calcium-activated potassium channel (BK) gene, called slowpoke. These cDNAs can be incorporated into different vectors for expression in cell lines or in Xenopus oocytes.
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Rat Strain Showing Developmental Changes in Alternative Splicing of the Heart Protein Titin

UW-Madison researchers have now discovered a mutant rat strain that shows an altered developmental pattern of titin expression. In these rats, the developmental transition in titin expression is dramatically retarded, so that titin in a 30-day old mutant rat is the same size as titin in a one-day old normal rat. These mutant animals should enable critical experiments to determine the effect of titin size on cardiac performance.
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Controllable Murine Models of First Pass Metabolism

UW-Madison researchers have developed mutant mouse lines that display an open (patent) ductus venosus. They also developed a way to control the closure of the ductus venosus so that some mice have first pass clearance while others do not. 

The researchers used embryonic stem cell targeting to create alleles of the transcriptional regulators AHR and ARNT with reduced gene expression. Because AHR and ARNT play a role in vascular development in the fetus, mutations affecting these genes alter blood vessel formation. The particular mutations developed by the researchers result in an open ductus venosus.
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Tsp1 -/- Stz Mice, A Model For Diabetes

Based on this observation, UW-Madison researchers have developed a line of TSP1-negative mice that provides an improved animal model for the study of diabetic retinopathy. They induced diabetes in TSP1-/- mice by injecting them with a single dose of streptozotocin to destroy their pancreatic beta cells. The resulting TSP1 -/- mice develop diabetes-associated early vasculopathies of similar severity to those observed in a previous mouse model developed by the inventors, but after a shorter duration of diabetes.
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A Mutation in the Rat Adenomatous Polyposis Coli Gene within the Human Mutation Hotspot Region

UW-Madison researchers have developed a strain of rats that more accurately recapitulates colon cancer in humans. They used N-ethyl-N-nitrosourea mutagenesis to create a truncating mutation in the rat APC gene at a site corresponding to the mutation hotspot region of the human ortholog. This germline mutation was confirmed by sequencing and has been transmitted to approximately half the offspring of the founder male. Unlike existing mouse models, these rats develop multiple adenomatous polyps in the colon, more faithfully imitating human colorectal cancer.
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Transgenic Zebrafish for Studying Neutrophil Chemotaxis and Inflammation

UW-Madison researchers have developed an animal model for studying neutrophil chemotaxis. They generated transgenic zebrafish that express enhanced green fluorescent protein (EGFP) through the myeloperoxide promoter. Myeloperoxide is a protein that neutrophils produce to kill bacteria and other pathogens. The EGFP labels the zebrafish neutrophils, allowing their location and movement to be detected using a microscope.
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Model for Stearoyl-CoA Desaturase Activity

UW-Madison researchers have developed a transgenic mouse model for the human SCD5 isoform. This animal model is useful for screening the activity and substrate specificity of human SCD5, and for identifying compounds that may modulate SCD5.
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Mouse Model of Glaucoma

A UW-Madison researcher has developed a mouse model that can be used for high throughput screening of neuroprotective glaucoma drugs. The retinal ganglion cells in these mice can easily be identified and assessed using a qualitative scoring system or image processing software.

To create the mouse model, the inventor started with DBA/2J mice, which develop secondary glaucoma. He then backcrossed an allele, known as R3, into these mice. R3 encodes a reporter protein that the mice express in their retinal ganglion cells.

To identify ganglion cells, the mice are euthanized, and their eyes are removed and stained for R3 protein activity. The retinas are then whole mounted onto glass slides and digitized. The ganglion cells appear as easily identifiable dark blue spots.

In addition, the inventor has developed a solution assay to quantitatively monitor the expression of the R3 gene. Because the silencing of this gene is part of the early stages of ganglion cell death, this assay makes it possible to evaluate whether certain drugs can potentially treat glaucoma before significant vision loss occurs.
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Mouse Strains and Cells Useful to Investigate Tissue-Specific Roles of Stearoyl-CoA Desaturase Isoforms

A UW–Madison researcher has developed several mouse strains and cells useful in the study of SCDs.  To understand the tissue-specific role of SCD1 and other SCD isoforms in disorders such as obesity, diabetes, cancer, inflammation, atherosclerosis and metabolism, the researcher generated mice with tissue-specific knockouts of SCDs.  He also developed transgenic mice in which SCDs can be overexpressed in specific tissues.
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Mouse Model for Mania

A UW–Madison researcher has developed a novel line of mice which exhibit at least four characteristics of mania: hyperactivity, elevated aggression, increased risk taking and decreased sleep.

The line of mice originally was derived from outbred hsd:ICR mice as part of a study of high wheel-running. The researcher now has found that the mice exhibit mania behaviors like decreased daytime sleep and cage hyperactivity. Furthermore, some of these behaviors were tempered following treatment with the common anti-mania drugs valproate and olanzapine, as well as lithium.
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Feline Model of Glaucoma

Researchers have identified a strain of cats with congenital glaucoma. They have captured the disease in a viable, outbred breeding colony with a highly informative pedigree. Results of molecular genetic and rigorous clinical and histopathologic studies confirm that the disease represents a homologue to a form of glaucoma in humans due to a mutation in LTBP2.
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Cryopreserved Head and Neck Cancer Tumorgrafts and Related Cell Strains

UW–Madison researchers have created a series of tumorgrafts from head and neck cancers, as well as cell lines from a subset of these tumorgrafts. Tumorgrafts are established by grafting human tumor samples directly onto mice and then passaging them onto other mice without ever placing them in tissue culture. They are expected to be more representative of actual human tumors than tumor xenograft mouse models.
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Wisconsin Miniature SwineTM for Biomedical Research

UW–Madison researchers have developed a novel line called “Wisconsin Miniature Swine,” or WMS, that is well suited to biomedical research and genetic modeling of human diseases. The animals’ body weight, size and composition are similar to humans and can be manipulated easily. For example, on an unrestricted feeding regimen, WMS become obese and appear to develop the hallmarks of metabolic syndrome.
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Wisconsin Miniature SwineTM for Cardiovascular Research

UW–Madison researchers have developed a novel line called Wisconsin Miniature Swine – Familial Hypercholesterolemia (WMS-FH). The animals are hypercholesterolemic and develop atherosclerotic vascular disease that is remarkably similar to that of humans.

The body weight, size and composition of WMS-FH are similar to humans and can be manipulated easily. They are ideally suited for studying the mechanisms of cardiovascular disease, and for developing/validating diagnostic and therapeutic technologies in the field of cardiology.
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Animal Model of Brittle Bone Disease

A UW–Madison researcher and collaborator have developed a mouse strain that is conditionally null for BMP1 and mTLL1. The model exhibits the hallmark features of OI and provides a new means to screen drugs and study mechanism.

The mice exhibit dramatically weakened and brittle bone with spontaneous fractures – defining features of OI. Additional skeletal features include osteomalacia, thinned/porous cortical bone, reduced processing of procollagen and dentin matrix protein 1, remarkably high bone turnover and defective osteocyte maturation.

WARF also owns and can license one of the parental lines (a mouse strain with a floxed TLL1 allele).
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Rat Model of Schizophrenia for Research and Preclinical Study

UW–Madison researchers have used a CRISPR/Cas9 approach to create several animal lines with deletions in exons 2/3 of Disc1, which has been identified as a significant candidate susceptibility gene in schizophrenia. Therefore the new model provides an exciting opportunity to explore the impact of this well-known de novo genetic variant on the structural and developmental features of the disease.
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Method to Screen for Novel Antibiotics

UW-Madison researchers have developed an indicator strain of Bacillus subtilis, which can detect potential antibiotic compounds that inhibit cell wall biosynthesis. This reporter strain includes the vancomycin-inducible VanA operon, which is responsible for a major form of inducible resistance to glycopeptide antibiotics, operably linked to a reporter gene.

To screen compounds for efficacy as antibiotics, a test compound is exposed to the strain. The effects of the compound on the indicator strain’s growth, as well as the presence or absence of the reporter gene product are observed. If the reporter gene product is present and growth of the indicator bacterial strain is inhibited or reduced, then the compound likely inhibits bacterial wall synthesis and is a potential, new antibiotic compound.
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