Project description:Retinal gene expression was assessed from postnatal day (P) 8 old low-density lipoprotein receptor-related protein 5 (Lrp5) knockout mice with a Illumina mouse-WG6 expression BeadChip.
Project description:Retinal gene expression was assessed from postnatal day (P) 8 old low-density lipoprotein receptor-related protein 5 (Lrp5) knockout mice with a Illumina mouse-WG6 expression BeadChip. RNA was extracted from the retinas of P8 Lrp5 knockout and wild type mice (n=3 each group). Total RNA (1ug each) were reverse transcribed, followed by a single in vitro transcription amplification to incorporate biotin-labeled nucleotide, and subsequent hybridization and staining with strepatavidin-Cy3 according to the manufacturer’s instructions.
Project description:The canonical Wnt signaling pathway has been demonstrated as a critical role in the self-renewal, proliferation and differentiation of Hematopoietic Stem Cells (HSCs), but the functions are indeterminacy in adult HSCs since the different experimental systems using gain- or loss- functions mice models. Low-density lipoprotein receptor-related proteins 5 and 6 (LRP5 and LRP6) are important co-receptors in the canonical Wnt/β-catenin pathway. In this study, the knockout mice models were established for adult HSC investagation via conditional ablation of LRP5 and LRP6 in adult hematopoiesis by Vav-Cre Loxp system. We found the HSC numbers were diminished due to the decreased proliferation and cell cycle. To investigate the molecular mechanisms, RNA-seq was performed using HSC cells isolated from WT and dKO mice.
Project description:To evaluate gene expression changes in response to low-density-lipoprotein related protein 1 (LRP1) depletion in endothelial cells, we performed mRNA-sequencing analysis with microvascular endothelial cells isolated from the liver LRP1 endothelial knockout mice and their littermate control wild type mice.
Project description:To evaluate gene expression changes in response to low-density-lipoprotein related protein 1 (LRP1) depletion in endothelial cells, we performed mRNA-sequencing analysis with microvascular endothelial cells isolated from the lung, heart and liver LRP1 endothelial knockout mice and their littermate control wild type mice.
Project description:Very low-density lipoprotein (VLDL) receptor (VLDLR), a member of the low-density lipoprotein receptor family, is responsible for VLDL uptake in peripheral tissues. We reported that significant increase in hepatic VLDLR levels following protein restriction in male mice does not contribute to hepatic fat accumulation, indicating that VLDLR may have hitherto unknown functions. Here, we used RNA-sequencing analysis of liver samples to analysis the effects of protein restriction on hepatic VLDLR in male and female mice.
Project description:Vision is essential for vertebrates including humans. Sustained vision is accomplished by retinoid metabolism, the ‘visual cycle’, where all-trans retinol (atROL) is incorporated into the retinal pigment epithelium (RPE) from photoreceptors presumably through decade-long missing receptor(s). Here, we show that the LDL-related receptor-5 (Lrp5) protein is linked to the retinol binding protein 1a (Rbp1a), the transporter of atROL in the visual cycle, by generating and analyzing the digenic eyes shut homolog+/-; lrp5+/- zebrafish, the same form of gene defect detected in a human case of inherited retinal degeneration. Global gene expression analysis followed by genetic study clarified that rbp1a played a role downstream of lrp5. Rbp1a protein was colocalized with Lrp5 protein at microvilli of RPE cells. Furthermore, Rbp1a directly bound to the C-terminal intracellular region of Lrp5 in vitro. Collectively, these results strongly suggest that Lrp5 is a potent candidate of the receptor of atROL in the visual cycle.
Project description:Emerging in vivo and vitro data suggest that white tea extract (WTE) is capable of favourably modulating metabolic syndrome, especially by ameliorating abnormal lipid metabolism. Microarray-based gene expression profiling was performed in HepG2 cells to analyze the effects of WTE from a systematic perspective. Gene Ontology and pathway analysis revealed that WTE significantly affected pathways related to lipid metabolism. WTE significantly downregulated apolipoprotein B (APOB) and microsomal triglyceride transfer protein (MTTP) expression and thereby reduced the production of very-low-density lipoprotein. In the meanwhile, WTE stimulated low-density lipoprotein-cholesterol (LDL-c) uptake through targeting low-density lipoprotein receptor (LDLR), as a consequence of the activation of sterol regulatory element-binding protein 2 (SREBP2) and peroxisome proliferator-activated receptor δ (PPARδ). Furthermore, WTE significantly downregulated triglycerides synthetic genes and reduced intracellular triglycerides accumulation. Besides, we demonstrated that the tea catechins epigallocatechin-3-gallate (EGCG) and epicatechin-3-gallate (ECG) are abundant in WTE and contribute to the regulation of cholesterol metabolism related genes, including LDLR, MTTP and APOB. Our findings suggest white tea plays important roles in ameliorating abnormal lipid metabolism in vitro.
Project description:Matrix metalloproteinase-12 (MMP12) is a proinflammatory protein secreted by macrophages. We have previously demonstrated that the genetic deletion of MMP12 in a cardiometabolic mouse model (low-density lipoprotein receptor-deficient (LdlrKO) mice fed a high-fat, sucrose- and cholesterol-enriched diet) ameliorated obesity-induced low-grade inflammation, white adipose tissue dysfunction, and atherosclerosis. Here, we hypothesized that the deletion of MMP12 might also positively affect whole-body energy metabolism and/or brown adipose tissue (BAT) function. Ldlr/Mmp12 double knockout (DKO) mice housed at room temperature (22°C) showed increased energy expenditure and decreased BAT size and triglyceride (TG) content. Untargeted proteomic analyses revealed the upregulation of several proteins and pathways related to mitochondrial function, glucose metabolism, and fatty acid oxidation in the BAT of DKO mice, whereas abundance of proteins and pathways associated with inflammation were reduced. In addition, DKO mice exhibited reduced macrophage infiltration in BAT with the infiltrating macrophages showing lower expression of lipid-associated marker genes. As we detected MMP12 expression in the mitochondrial fraction of BAT, its absence in DKO BAT was consistent with reduced oxygen consumption, compactness, and sphericity of the mitochondria. Following an acute cold exposure, DKO mice had decreased BAT weights and circulating lipid concentrations, especially very low-density lipoprotein-TG and LDL-cholesterol, and increased expression of thermogenic genes. We conclude that MMP12 may play a detrimental role in whole-body energy homeostasis and thermogenesis, as it triggers macrophage infiltration, inflammation, and mitochondria dysfunction in BAT.