Project description:Previous studies in our laboratory have shown that low folate diet (control diet with 2mg folate/kg, low folate diet with 0.3mg folate/kg) can induce intestinal tumors in BALB/c mice. In addition, we reported that C57Bl/6J mice did not form tumors under the same conditions. We used microarrays to identify the genetic differences between BALB/c and C57Bl/6J mice that promote tumorigenesis in BALB/c mice.
Project description:Soleus muscle CAGE profiles of mice (C57BL/6J) under normal conditions, torpid by fasting, under high ambient temperature (32°C), torpor deprived were generated by deep sequencing, in two runs and 2-8 replicates, using Illumina NextSeq.
Project description:Natural variation in protein expression is common in all organisms and contribute to phenotypic differences among individuals. While variation in gene expression at the transcript level has been extensively investigated, the genetic mechanisms underlying variation in protein expression have lagged considerably behind. Here we investigate genetic architecture of protein expression by profiling a deep mouse brain proteome of two inbred strains, C57BL/6J (B6) and DBA/2J (D2), and their reciprocal F1 hybrids using two-dimensional liquid chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) technology. By comparing protein expression levels in the four mouse strains, we observed 329 statistically significant differentially expressed proteins between the two parental strains and identified four common inheritance patterns, including dominant, additive, over- and under-dominant expression. We further applied the proteogenomic approach to detect variant peptides and define protein allele-specific expression (pASE).
Project description:Study designed to identify genes that are induced in both BAT and skeletal muscle during acute adaptive thermogenesis in mouse using gene expression microarray. Animal studies were performed under approved UCLA animal research protocols and according to guidelines established in the Guide for Care and Use of Laboratory Animals. C57BL/6J mice were maintained in 12-h light/dark conditions and fed a regular chow diet (Purina 50010, Lab Diet, USA). Cold exposure was performed as described. After euthanasia, tissues were collected and frozen until use. Total RNA was isolated from mouse tissues by extraction with TRIzol from either brown adipose tissue (BAT) or skeletal muscle (quadriceps) after exposure to cold (4 degrees C for 4 hours). The mice were of the C57BL/6J strain and were maintained in 12-h light/dark conditions and fed a laboratory chow diet that consisted of 4.5% fat, 50% carbohydrate by weight (Lab Diet, Purina 50010).
Project description:Co-immunoprecipitation with either rabbit normal IgG or anti-Nogo-B antibody followed by mass spectrometry (co-IP-MS) in the liver and pancreas of wild-type C57BL/6J mice.
Project description:Endoplasmic reticulum (ER) stress occurs when misfolded proteins accumulate in the ER. The cellular response to ER stress involves complex transcriptional and translational changes, important to the survival of the cell. ER stress is a primary cause and a modifier of many human diseases. A first step to understanding how the ER stress response impacts human disease is to determine how the transcriptional response to ER stress varies among individuals. The genetic diversity of the eight mouse Collaborative Cross (CC) founder strains allowed us to determine how genetic variation impacts the ER stress transcriptional response. We used tunicamycin, a drug commonly used to induce ER stress, to elicit an ER stress response in mouse embryonic fibroblasts (MEFs) derived from the CC founder strains and measured their transcriptional responses. We identified hundreds of genes that differed in response to ER stress across these genetically diverse strains. Strikingly, inflammatory response genes differed most between strains; major canonical ER stress response genes showed relatively invariant responses across strains. To uncover the genetic architecture underlying these strain differences in ER stress response, we measured the transcriptional response to ER stress in MEFs derived from a subset of F1 crosses between the CC founder strains. We found a unique layer of regulatory variation that is only detectable under ER stress conditions. Over 80% of the regulatory variation under ER stress derives from cis-regulatory differences. This is the first study to characterize the genetic variation in ER stress transcriptional response in the laboratory mouse. Our findings indicate that the ER stress transcriptional response is highly variable among strains and arises from genetic variation in individual downstream response genes, rather than major signaling transcription factors. These results have important implications for understanding how genetic variation impacts the ER stress response, an important component of many human diseases. We investigated the genetic variation in ER stress transcriptional response in mouse embryonic fibroblasts (MEFs) across eight mouse strains: A/J, C57BL/6J, 129S1Sv/ImJ, NOD/ShiLtJ, NZO/H1LtJ, CAST/EiJ, PWK/PhJ, and WSB/EiJ. MEFs from each strain were treated with a control DMSO or ER stress-inducing drug, Tunicamycin (TM). To identify the genetic architecture underlying this genetic variation, MEFs from F1 strains were also studied. MEFs from the following F1s were evaluated: C57BL/6J X CAST/EiJ, C57BL/6J X 129S1Sv/ImJ, C57BL/6J X NOD/ShiLtJ, C57BL/6J X NZO/H1LtJ, and C57BL/6J X WSB/EiJ. Again F1 MEFS were treated with either DMSO or TM. There are two or three replicates for each sample.
Project description:Eight weeks old male Adrb1tm1BkkAdrb2tm1Bkk/J , stock number 003810, were purchased from The Jackson Laboratory. These mice are homozygous null for the Adrb1 and Adrb2 genes, and are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Mice were euthanized, and the whole bone marrow was extracted using established methods.1 Whole bone marrow cells from Adrb1tm1BkkAdrb2tm1Bkk/J mice were reconstituted into lethally- irradiated (950 Rad) C57BL/6J mice using a single retro-orbital injection at a ratio of 1:4 (Adrb1tm1BkkAdrb2tm1Bkk/J to C57BL/6J). All reconstituted mice were recovered for 2-3 months prior to tissue collection for mRNA arrays (1) The FASEB Journal 2015:29(1),652.13.
Project description:Eight weeks old male Adrb1tm1BkkAdrb2tm1Bkk/J , stock number 003810, were purchased from The Jackson Laboratory. These mice are homozygous null for the Adrb1 and Adrb2 genes, and are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Mice were euthanized, and the whole bone marrow was extracted using established methods.1 Whole bone marrow cells from Adrb1tm1BkkAdrb2tm1Bkk/J mice were reconstituted into lethally- irradiated (950 Rad) C57BL/6J mice using a single retro-orbital injection at a ratio of 1:4 (Adrb1tm1BkkAdrb2tm1Bkk/J to C57BL/6J). All reconstituted mice were recovered for 2-3 months prior to tissue collection for mRNA arrays (1) The FASEB Journal 2015:29(1),652.13.