Project description:NIH3T3 cell transiently transfected with myocardin-EGFP or EGFP. Transfection: Lipofectamine 2000 according to the manufacturers instructions (Invitrogen).
Project description:Effect of the loss of both miR-1/133a clusters on the embryonic heart. Please note: dKO means it is a double KO of the two miR-1/133a clusters in the mouse. Control means we do not use just WT but also some heterozygous animals to compare to dKOs, however these controls do not have a phenotype. Transgene negative: these are littermates of the myocd transgenic animals that do not carry a transgene.
Project description:Inhibition of myostatin signaling induces strong skeletal muscle growth making it an attractive target to treat muscle wasting and sarcopenia. However, the biological function of myostatin in the heart is barely understood. We demonstrate that conditional inactivation of myostatin in the adult murine heart leads to cardiac hypertrophy, heart failure and increased lethality. To induce cardiomyocyte specific loss of myostatin a conditionally active Mstn^fl/fl allele was generated by insertion of loxP elements upstream and downstream of exons 1 and 2 of the mouse myostatin gene. The selection cassette was removed in vivo by flp-recombination. To inactivate myostatin, mice were mated to alphaMyHC-MCM mice (Sohal, DS, et al. (2001) Circulation Research 89, 20-25). Cre-recombination was achieved by intraperitoneal administration of Tamoxifen (40 mg/kg) for 5 consecutive days. The respective control alphaMyHC-MCM animals were equally treated.
Project description:The ventricular wall of the heart is composed of trabeculated and compactlayers, which are separated by yet unknown processes during embryonic development. Notch signaling plays important roles in cardiac development. Mutations in Notch signaling components are related to human congenital heart diseases Two homologues, Numb and Numblike (Numbl), are expressed in mammals, which also act as inhibitors of Notch signaling Here we investigate the role of Notch2 and Numb/Numblike during myocardial trabeculation and compaction. The aim of the experiment is to analyze whether Numb/Numblike are involved in inhibition of Notch activity in the developing heart.
Project description:Autophagy is a lysosomal degradation process involved in the turnover of organelles or other cell constituents, in providing sources for energy production under starving conditions, and in cell metabolism. A key protein in the macroautophagic machinery is the autophagy related protein (Atg) 7. Constitutive deletion of Atg7 is lethal at birth. A conditional deletion of Atg7 in hepatocytes leads to hepatomegaly and in aged animals to liver tumors. With this study, we aimed at analyzing the hepatomegaly development more detailed. The 3- to 4-fold enlargement of the liver takes place between day 25 and 35 after birth (P25-P35) and persists at least until P90. This is accompanied by a change in the expression of enzymes involved in the glycogen/glucose metabolism. While glycogen synthesis is inhibited, glucose is preferentially kept as glucose-6-phosphate inside the cells, inducing a swelling of the cells caused by hyperosmolarity. An increase of lipogenic enzymes suggests that glucose-6-phosphate is delivered to lipogenic pathways, which is supported by the occurrence of a steatosis around P30. The development of hepatomegaly is accompanied by a polyploidisation of hepatocytes, an enhanced expression of genes related to inflammatory processes, and an infiltration of macrophages. Our data provide evidence that the attenuation of macroautophagy in hepatocytes leads to a glucose retention which causes cell swelling. The resulting hepatomegaly, which develops in a time interval of about 10 days, perturbs liver perfusion and induces an inflammatory reaction together with polyploidisation.
Project description:In this project, we identified a novel RNA-binding protein, MHZ9. And we analyzed the potential proteins interacted with MHZ9 through immunoprecipitation-mass spectrometry (IP-MS). The N-terminal domain of MHZ9 (MHZ9-N) contains a putative RNA splicing and modification domain PRP4. To identify RNA binding sites in the MHZ9-N. We performed XRNAX-IP-MS assay.