Project description:To determine the effect of insulin deficiency on global changes to expression of glycan related genes in the liver. See: Joseph R. Bishop,‡ Erin Foley,‡§ Roger Lawrence,‡ and Jeffrey D. Esko‡1 (2010) Insulin-dependent Diabetes Mellitus in Mice Does Not Alter Liver Heparan Sulfate* J Biol Chem. 2010 May 7; 285(19): 14658–14662. We are studying the glycan changes caused by insulin-deficient diabetes in the mouse liver. We would like to determine whether insulin-deficiency causes global changes to expression of glycan-related genes in the liver. Male C57BL/6 mice (4 weeks of age) were purchased from Jackson Laboratory and maintained in a temperature-controlled (25 °C) facility with a 12-h light/dark cycle. The derivation and genotyping of Ndst1f/fAlbCre+ mice have been described previously (16). Mice were fed laboratory rodent chow (Harlan-Teklad) ad libitum except when fasting blood specimens were obtained. Mice were made diabetic by administering 50 mg/kg body weight of streptozotocin (STZ; Sigma) intraperitoneally for 5 consecutive days. Because of variations in plasma triglycerides in females, only male mice were used in this study. Our preliminary results suggest glycosaminoglycan enzymes may be down-regulated, which may affect the turnover of lipids in the plasma. Liver RNA samples from 3 diabetic mice will be compared to 3 wildtype samples for properly controlled analysis.
Project description:To determine the effect of insulin deficiency on global changes to expression of glycan related genes in the liver. See: Joseph R. Bishop,‡ Erin Foley,‡§ Roger Lawrence,‡ and Jeffrey D. Esko‡1 (2010) Insulin-dependent Diabetes Mellitus in Mice Does Not Alter Liver Heparan Sulfate* J Biol Chem. 2010 May 7; 285(19): 14658–14662. We are studying the glycan changes caused by insulin-deficient diabetes in the mouse liver. We would like to determine whether insulin-deficiency causes global changes to expression of glycan-related genes in the liver. Male C57BL/6 mice (4 weeks of age) were purchased from Jackson Laboratory and maintained in a temperature-controlled (25 °C) facility with a 12-h light/dark cycle. The derivation and genotyping of Ndst1f/fAlbCre+ mice have been described previously (16). Mice were fed laboratory rodent chow (Harlan-Teklad) ad libitum except when fasting blood specimens were obtained. Mice were made diabetic by administering 50 mg/kg body weight of streptozotocin (STZ; Sigma) intraperitoneally for 5 consecutive days. Because of variations in plasma triglycerides in females, only male mice were used in this study.
Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from Mus musculus tissues (Heart, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:Metabolic abnormalities underlying diabetes are primarily the result of the lack of adequate insulin action and the associated changes in protein phosphorylation and gene expression. Affymetrix oligonucleotide microarrays were used to study the changes in the transcriptional program of mouse skeletal muscle in insulin-deficient diabetes. Mice which were made diabetic by streptozotocin treatment were compared to controls. Also, the reversibility of these changes was ascertained by treating a subset of the diabetic mice with insulin.
Project description:To evaluate functional consequences of insulin-deficient diabetes mellitus for the liver, we used a genetically engineered pig model of mutant INS gene induced diabetes of youth (MIDY). Liver samples of MIDY pigs and wild-type (WT) littermate controls were analyzed by label-free proteomics to reveal pathways and key drivers significantly affected by chronic insulin deficiency and hyperglycemia.