Articles by Timothy L. Hostelley in JoVE
Sample Preparation and Analysis of RNASeq-based Gene Expression Data from Zebrafish Timothy L. Hostelley*1, Jessica E. Nesmith*1, Norann A. Zaghloul1 1Division of Endocrinology, Diabetes and Nutrition, Department of Medicine, University of Maryland School of Medicine This protocol presents an approach for whole transcriptome analysis from zebrafish embryos, larvae, or sorted cells. We include isolation of RNA, pathway analysis of RNASeq data, and qRT-PCR-based validation of gene expression changes.
Other articles by Timothy L. Hostelley on PubMed
Differential Effects on β-cell Mass by Disruption of Bardet-Biedl Syndrome or Alstrom Syndrome Genes Human Molecular Genetics. Jan, 2016 | Pubmed ID: 26494903 Rare genetic syndromes characterized by early-onset type 2 diabetes have revealed the importance of pancreatic β-cells in genetic susceptibility to diabetes. However, the role of genetic regulation of β-cells in disorders that are also characterized by highly penetrant obesity, a major additional risk factor, is unclear. In this study, we investigated the contribution of genes associated with two obesity ciliopathies, Bardet-Biedl Syndrome and Alstrom Syndrome, to the production and maintenance of pancreatic β-cells. Using zebrafish models of these syndromes, we identified opposing effects on production of β-cells. Loss of the Alstrom gene, alms1, resulted in a significant decrease in β-cell production whereas loss of BBS genes, bbs1 or bbs4, resulted in a significant increase. Examination of the regulatory program underlying β-cell production suggested that these effects were specific to β-cells. In addition to the initial production of β-cells, we observed significant differences in their continued maintenance. Under prolonged exposure to high glucose conditions, alms1-deficient β-cells were unable to continually expand as a result of decreased proliferation and increased cell death. Although bbs1-deficient β-cells were similarly susceptible to apoptosis, the overall maintenance of β-cell number in those animals was sustained likely due to increased proliferation. Taken together, these findings implicate discrepant production and maintenance of β-cells in the differential susceptibility to diabetes found between these two genetic syndromes.
Whole Organism Transcriptome Analysis of Zebrafish Models of Bardet-Biedl Syndrome and Alström Syndrome Provides Mechanistic Insight into Shared and Divergent Phenotypes BMC Genomics. May, 2016 | Pubmed ID: 27142762 Bardet-Biedl Syndrome (BBS) and Alström Syndrome are two pleiotropic ciliopathies with significant phenotypic overlap between them across many tissues. Although BBS and Alström genes are necessary for the proper function of primary cilia, their role in defects across multiple organ systems is unclear.