Gene expression in the embryonic Lrrc10 knockout heart
Ontology highlight
ABSTRACT: LRRC10 is a heart-specific gene required for proper cardiac function. The effects of Lrrc10 deletion on gene expression in the embryonic mouse heart was investigated. Pregnant heterozygous Lrrc10 knockout mice (Lrrc10+/-) mated to male heterozygous knockouts (Lrrc10+/-), housed in 12 hour light:12 dark, ad lib feeding and drinking conditions, were sacrificed at embryonic day 15.5 (E15.5) and embryonic hearts were dissected for cardiac gene expression analysis. A two color, reference design experiment in which heart RNA from 2 Lrrc10 homozygous knockout embryos (Lrrc10-/-) were pooled and labeled with Cy5 and hybridized according to Agilent protocols against a reference pool of RNA madeup from respective tissue taken from littermate E15.5 wildtype (Lrrc10+/+) hearts and labeled with Cy3.
Project description:LRRC10 is a heart-specific gene required for proper cardiac function. The effects of Lrrc10 deletion on gene expression in the adult mouse heart was investigated. Lrrc10 knockout mice or wildtype controls, housed in 12 hour light:12 dark, ad lib feeding and drinking conditions were sacrificed at two months of age for cardiac gene expression analysis. A two color, reference design experiment in which heart RNA from 2 Lrrc10 knockout mice was pooled and labeled with Cy5 and hybridized according to Agilent protocols against a reference pool of RNA madeup from respective tissue taken from 2 month wildtype mice which was labeled with Cy3.
Project description:The diurnal variation in acetaminophen (APAP) hepatotoxicity (“chronotoxicity”) is thought to be due to oscillations in xenobiotic metabolism that are influenced by the circadian phases of feeding or fasting. Because of APAP’s relevance to human poisoning, we set out to determine the relative contributions of the central clock in the SCN and the autonomous clock in the hepatocyte in modulating the chronotoxicity of APAP. Using a conditional null allele of Mop3 (ArntL, Bmal1) we are able to delete the clock from hepatocytes while keeping the central and other peripheral clocks intact (eg, those controlling food intake). Our data from this hepatocyte-null mouse model suggests that, while the central circadian clock modulates some detoxification pathways indirectly by driving activity patterns and feeding rhythms, the autonomous hepatocyte circadian clock controls major aspects of APAP bioactivation independent of feeding rhythms, possibly through transcriptional regulation of cytochrome p450-oxidoreductase (Por). 10-20 week old Mop3fxfx mice positive or negative for Cre-recombinase driven by the albumin promoter, housed in 12 hour light:12 dark, ad lib feeding and drinking conditions were sacrificed every four hours over two separte days beginning at ZT0. A two color, reference design experiment in which kidney RNA from at least 3 mice per timepoint were pooled and labeled with Cy3 and hybridized according to Agilent protocols against a reference pool of RNA madeup from respective tissue taken from 10 week Mop3fxfx and Mop3fxfxCreAlb mice which was labeled with Cy5.
Project description:The diurnal variation in acetaminophen (APAP) hepatotoxicity (“chronotoxicity”) is thought to be due to oscillations in xenobiotic metabolism that are influenced by the circadian phases of feeding or fasting. Because of APAP’s relevance to human poisoning, we set out to determine the relative contributions of the central clock in the SCN and the autonomous clock in the hepatocyte in modulating the chronotoxicity of APAP. Using a conditional null allele of Mop3 (ArntL, Bmal1) we are able to delete the clock from hepatocytes while keeping the central and other peripheral clocks intact (eg, those controlling food intake). Our data from this hepatocyte-null mouse model suggests that, while the central circadian clock modulates some detoxification pathways indirectly by driving activity patterns and feeding rhythms, the autonomous hepatocyte circadian clock controls major aspects of APAP bioactivation independent of feeding rhythms. 10-20 week old Mop3fxfx mice positive or negative for Cre-recombinase driven by the albumin promoter, housed in 12 hour light:12 dark, ad lib feeding and drinking conditions were sacrificed every four hours over two separte days beginning at ZT0. A two color, reference design experiment in which liver RNA from at least 3 mice per timepoint were pooled and labeled with Cy3 and hybridized according to Agilent protocols against a reference pool of RNA madeup from respective tissue taken from 10 week Mop3fxfx and Mop3fxfxCreAlb mice which was labeled with Cy5.
Project description:Transcriptional profiling of mouse whole liver comparing control WT B6 mice with B6 growth hormone-deficient little, B6 androgen receptor-null Tfm mice, and STAT5b KOs normalized to WT on B6 and BALB/c backgrounds. All animals were 10-week-old males initiated with DEN. Oberley et al. Molecular carcinogenesis 2014 May 17. doi: 10.1002/mc.22165. Three-condition experiment, WT vs. little and Tfm livers. Biological replicates: 3 control replicates, 3 mutant replicates. Little and Tfm normalized to WTa, B6 STAT5b KO
Project description:Dextran sodium sulfate (DSS) causes inflammation in the gut similar to ulcerative colitis in humans. Patients with ulcerative colitis have increased risk of developing colon cancer. We sought to determine which genes are altered in the normal colonic epithelium, and which changes depend on the Pirc mutation. 97 day old (ACIxF344)F1 wild type and Pirc male rats either untreated or given 4% DSS in the drinking water from 40-47 and 54-61 days of age, housed in 12 hour light:12 dark, ad lib feeding. Normal colonic tissue was collected from the distal colon at 97 days of age.
Project description:Pathogenic variants in ACTN2, coding for alpha-actinin 2, are known to be rare causes of Hyper-trophic Cardiomyopathy. However, little is known about the underlying disease mechanisms. Adult heterozygous mice carrying the Actn2 M228T variant were phenotyped by echocardiog-raphy. For homozygous mice, viable E15.5 embryonic hearts were analysed by High Resolution Episcopic Microscopy and wholemount staining, complemented by unbiased proteomics, qPCR and Western blotting. Heterozygous Actn2 M228T mice have no overt phenotype. Only mature males show molecular parameters indicative of cardiomyopathy. By contrast, the variant is em-bryonically lethal in the homozygous setting and E15.5 hearts show multiple morphological ab-normalities. Molecular analyses, including unbiased proteomics, identified quantitative abnormal-ities in sarcomeric parameters, cell cycle defects and mitochondrial dysfunction. The mutant al-pha-actinin protein is found to be destabilised, associated with increased activity of the ubiqui-tin-proteosomal system. This missense variant in alpha-actinin renders the protein less stable. In response, the ubiquitin-proteosomal system is activated; a mechanism which has been implicated in cardiomyopathies previously. In parallel, lack of functional alpha-actinin is thought to cause energetic defects through mitochondrial dysfunction. This seems, together with cell cycle defects, the likely cause of death of the embryos. The defects also have wide-ranging morphological con-sequences.
Project description:Cardiac remodelling is a key process in the development of heart failure. Reactivation of foetal cardiac genes is often associated with cardiac remodelling, which might be the result of the activation of signalling pathways involved in heart failure. Here we studied the role of Pontin (Ruvbl1), which is highly expressed in embryonic hearts, in mediating adverse remodelling in adult mouse hearts.
Project description:We have recently shown a remarkable regenerative capacity of the prenatal heart using a genetic model of mosaic mitochondrial dysfunction in mice. This model is based on inactivation of the X-linked gene encoding holocytochrome c synthase (Hccs) specifically in the developing heart. Loss of HCCS activity results in respiratory chain dysfunction, disturbed cardiomyocyte differentiation and reduced cell cycle activity. The Hccs gene is subjected to X chromosome inactivation, such that in females heterozygous for the heart conditional Hccs knockout approximately 50% of cardiac cells keep the defective X chromosome active and develop mitochondrial dysfunction while the other 50% remain healthy. During heart development, however, the contribution of HCCS deficient cells to the cardiac tissue decreases from 50% at midgestation to 10% at birth. This regeneration of the prenatal heart is mediated by increased proliferation of the healthy cardiac cell population, which compensate for the defective cells and allow the formation of a fully functional heart at birth. Here we performed microarray expression ananlyses on 13.5 dpc control and heterozygous Hccs knockout hearts to identify molecular mechanisms that drive embryonic heart regeneration.