Project description:We aim to analyze the transcriptional profiles of keratinocytes near dermal-epidemal junction during HSV-2 reactivation in humans, including lesion, post healed (4 and 8 weeks post healing) and contra-lateral control biopsies. Since keratinocytes are the main target cells for HSV-2 infection, the goal is to define the intrinsic immunity of keratinocytes during HSV-2 reactivation in humans.
Project description:Deficiency in the protein-tyrosine phosphatase SHP1/PTPN6 is linked with hematological malignancies and chronic inflammatory diseases. Here we exploited the embryonic and larval stages of zebrafish as an animal model to study ptpn6 function in the sole context of innate immunity. We show that ptpn6 knockdown induces a spontaneous inflammation-associated phenotype at the late larval stage, which was microbe-independent and enhanced instead of suppressed by glucocorticoids. When challenged with Salmonella typhimurium or Mycobacterium marinum at earlier stages of development, the innate immune system was hyperactivated to a contra-productive level that impaired the control of these pathogenic bacteria. These results demonstrate the crucial regulatory function of ptpn6 in preventing host-detrimental effects of inflammation by imposing a tight control over the level of innate immune response activation.
Project description:Website for easy exploration of the data: https://rna-seq-browser.herokuapp.com/# For the first time, we here characterize the transcriptional signature of satellite glial cells (SGCs) in a nerve injury-paradigm by isolating SGCs from mouse dorsal root ganglia (DRGs) after peripheral nerve injury followed by next generation RNA sequencing (RNA-seq). We found that SGCs regulate their gene expression differently at 3 days and 14 days after peripheral nerve injury suggesting that they change their function over time. At both time points, we found downregulation of several genes linked to cholesterol. After 14 days we further detect regulation of genes linked to the immune system (MHC protein complex and leukocyte migration). The SGC RNA-seq data is accompanied by RNA-seq of purified nociceptors from injured (partial sciatic nerve ligation) or sham (ipsi after sham operation or contra after partial sciatic nerve ligation) L3-L5 DRGs.
Project description:CREB-Regulated Transcription Co-activator (CRTC) regulates metabolism in liver where activation by calcineurin regulates gluconeogenic genes. CaN also has roles in pathological cardiac hypertrophy, however cardiac roles for CRTC have not been identified. In Drosophila, CRTC null mutants exhibit severe cardiac restriction, myofibrillar disorganization, cardiac fibrosis, and tachycardia. Cardiac-specific knockdown (KD) of CRTC mimicked the heart defects of CRTC mutants and cardiac-overexpression (OE) of CRTC or calcineurin caused hypertrophy that was reduced in CRTC mutants, suggesting CRTC mediates calcineurin’s effects. RNAseq of CRTC KD or OE hearts revealed contra-regulated genes involved in glucose, fatty acid, and amino acid metabolism. Genes with conserved CREB binding sites included the fly ortholog of Sarcalumenin, a Ca2+-binding protein. Cardiac KD of this gene recapitulated CRTC KD restriction and fibrotic phenotypes. KD in zebrafish also caused restriction, indicating a conserved role in cardiomyocyte maintenance, and suggesting CaN-CRTC-Sarcalumenin signaling represents a novel pathway underlying cardiac hypertrophy.
Project description:The initial segment of the epididymis is vital for male fertility, therefore, it is important to understand the mechanisms that regulate this important region. Deprival of testicular luminal fluid factors/lumicrine factors from epididymis, a subset of cells within the initial segment undergo apoptosis. In this study, microarray analyses was used to examine early changes in the downstream signal transduction pathways following the loss of lumicrine factors, and we discovered the following cascade of events leading to loss of protection and eventual apoptosis. First, mRNA expression of several key components of ERK pathway decreased sharply after 6 hours of loss protection from testicular lumicrine factors. After 12 hours, the levels of mRNA expression of STAT and NF-кB pathways components increased, mRNA expression of genes encoding cell cycle inhibitors increased. After 18 hours of loss protection from testicular lumicrine factors, apoptosis was observed in the initial segment. In conclusion, testicular lumicrine factors protect the cells of the initial segment by activating ERK pathway, repressing STAT and NF-кB pathways, and preventing a cascade of reactions leading to apoptosis. Unilateral efferent duct ligation (EDL) surgeries were performed to block lumicrine testicular fluid factors from reaching one side of epididymis. For the control, a sham operation was performed on the contra-lateral side within the same animal. After 6, 12 and 18 hours of EDL, the most proximal region of the initial segment containing zone 1a and 1b of initial segment was dissected out. In this microarray study, sham control and each time point of EDL (sham, 6h EDL, 12h EDL, 18h EDL) all have 4 replicates, a total of 16 arrays were hybridized.