96 research outputs found
Genome-Wide Tissue-Specific Occupancy of the Hox Protein Ultrabithorax and Hox Cofactor Homothorax in Drosophila
The Hox genes are responsible for generating morphological diversity along the
anterior-posterior axis during animal development. The
Drosophila Hox gene Ultrabithorax
(Ubx), for example, is required for specifying the identity
of the third thoracic (T3) segment of the adult, which includes the dorsal
haltere, an appendage required for flight, and the ventral T3 leg.
Ubx mutants show homeotic transformations of the T3 leg
towards the identity of the T2 leg and the haltere towards the wing. All Hox
genes, including Ubx, encode homeodomain containing
transcription factors, raising the question of what target genes
Ubx regulates to generate these adult structures. To
address this question, we carried out whole genome ChIP-chip studies to identify
all of the Ubx bound regions in the haltere and T3 leg imaginal discs, which are
the precursors to these adult structures. In addition, we used ChIP-chip to
identify the sites bound by the Hox cofactor, Homothorax (Hth). In contrast to
previous ChIP-chip studies carried out in Drosophila embryos,
these binding studies reveal that there is a remarkable amount of tissue- and
transcription factor-specific binding. Analyses of the putative target genes
bound and regulated by these factors suggest that Ubx regulates many downstream
transcription factors and developmental pathways in the haltere and T3 leg.
Finally, we discovered additional DNA sequence motifs that in some cases are
specific for individual data sets, arguing that Ubx and/or Hth work together
with many regionally expressed transcription factors to execute their functions.
Together, these data provide the first whole-genome analysis of the binding
sites and target genes regulated by Ubx to specify the morphologies of the adult
T3 segment of the fly
Oxygen-sensing neurons reciprocally regulate peripheral lipid metabolism via neuropeptide signaling in <i>Caenorhabditis elegans</i>
<div><p>The mechanisms by which the sensory environment influences metabolic homeostasis remains poorly understood. In this report, we show that oxygen, a potent environmental signal, is an important regulator of whole body lipid metabolism. <i>C</i>. <i>elegans</i> oxygen-sensing neurons reciprocally regulate peripheral lipid metabolism under normoxia in the following way: under high oxygen and food absence, URX sensory neurons are activated, and stimulate fat loss in the intestine, the major metabolic organ for <i>C</i>. <i>elegans</i>. Under lower oxygen conditions or when food is present, the BAG sensory neurons respond by repressing the resting properties of the URX neurons. A genetic screen to identify modulators of this effect led to the identification of a BAG-neuron-specific neuropeptide called FLP-17, whose cognate receptor EGL-6 functions in URX neurons. Thus, BAG sensory neurons counterbalance the metabolic effect of tonically active URX neurons via neuropeptide communication. The combined regulatory actions of these neurons serve to precisely tune the rate and extent of fat loss to the availability of food and oxygen, and provides an interesting example of the myriad mechanisms underlying homeostatic control.</p></div
IL-17 Expression in the Time Course of Acute Anti-Thy1 Glomerulonephritis
Background Interleukin-17 (IL-17) is a new pro-inflammatory cytokine involved
in immune response and inflammatory disease. The main source of IL-17 is a
subset of CD4+ T-helper cells, but is also secreted by non-immune cells. The
present study analyzes expression of IL-17 in the time course of acute anti-
thy1 glomerulonephritis and the role of IL-17 as a potential link between
inflammation and fibrosis. Methods Anti-thy1 glomerulonephritis was induced
into male Wistar rats by OX-7 antibody injection. After that, samples were
taken on days 1, 5, 10 (matrix expansion phase), 15 and 20 (resolution phase).
PBS-injected animals served as controls. Proteinuria and histological matrixes
score served as the main markers for disease severity. In in vitro
experiments, NRK-52E cells were used. For cytokine expressions, mRNA and
protein levels were analyzed by utilizing RT-PCR, in situ hybridization and
immunofluorescence. Results Highest IL-17 mRNA-expression (6.50-fold vs. con;
p<0.05) was found on day 5 after induction of anti-thy1 glomerulonephritis
along the maximum levels of proteinuria (113 ± 13 mg/d; p<0.001), histological
glomerular-matrix accumulation (82%; p<0.001) and TGF-β1 (2.2-fold; p<0.05),
IL-6 mRNA expression (36-fold; p<0.05). IL-17 protein expression co-localized
with the endothelial cell marker PECAM in immunofluorescence. In NRK-52E
cells, co-administration of TGF-β1 and IL-6 synergistically up-regulated IL-17
mRNA 4986-fold (p<0.001). Conclusions The pro-inflammatory cytokine IL-17 is
up-regulated in endothelial cells during the time course of acute anti-thy1
glomerulonephritis. In vitro, NRK-52E cells secrete IL-17 under pro-fibrotic
and pro-inflammatory conditions
Pollen and Phytoliths from Fired Ancient Potsherds as Potential Indicators for Deciphering Past Vegetation and Climate in Turpan, Xinjiang, NW China
It is demonstrated that palynomorphs can occur in fired ancient potsherds when the firing temperature was under 350°C. Pollen and phytoliths recovered from incompletely fired and fully fired potsherds (ca. 2700 yrs BP) from the Yanghai Tombs, Turpan, Xinjiang, NW China can be used as potential indicators for reconstructing past vegetation and corresponding climate in the area. The results show a higher rate of recovery of pollen and phytoliths from incompletely fired potsherds than from fully fired ones. Charred phytoliths recovered from both fully fired and incompletely fired potsherds prove that degree and condition of firing result in a permanent change in phytolith color. The palynological data, together with previous data of macrobotanical remains from the Yanghai Tombs, suggest that temperate vegetation and arid climatic conditions dominated in the area ca. 2700 yrs BP
OrthoList: A Compendium of C. elegans Genes with Human Orthologs
C. elegans is an important model for genetic studies relevant to human biology and disease. We sought to assess the orthology between C. elegans and human genes to understand better the relationship between their genomes and to generate a compelling list of candidates to streamline RNAi-based screens in this model.We performed a meta-analysis of results from four orthology prediction programs and generated a compendium, "OrthoList", containing 7,663 C. elegans protein-coding genes. Various assessments indicate that OrthoList has extensive coverage with low false-positive and false-negative rates. Part of this evaluation examined the conservation of components of the receptor tyrosine kinase, Notch, Wnt, TGF-ß and insulin signaling pathways, and led us to update compendia of conserved C. elegans kinases, nuclear hormone receptors, F-box proteins, and transcription factors. Comparison with two published genome-wide RNAi screens indicated that virtually all of the conserved hits would have been obtained had just the OrthoList set (∼38% of the genome) been targeted. We compiled Ortholist by InterPro domains and Gene Ontology annotation, making it easy to identify C. elegans orthologs of human disease genes for potential functional analysis.We anticipate that OrthoList will be of considerable utility to C. elegans researchers for streamlining RNAi screens, by focusing on genes with apparent human orthologs, thus reducing screening effort by ∼60%. Moreover, we find that OrthoList provides a useful basis for annotating orthology and reveals more C. elegans orthologs of human genes in various functional groups, such as transcription factors, than previously described
Differential gene and protein expression of chemokines and cytokines in synovial fluid of patients with arthritis
Insights into Hox Protein Function from a Large Scale Combinatorial Analysis of Protein Domains
Protein function is encoded within protein sequence and protein domains. However, how protein domains cooperate within a protein to modulate overall activity and how this impacts functional diversification at the molecular and organism levels remains largely unaddressed. Focusing on three domains of the central class Drosophila Hox transcription factor AbdominalA (AbdA), we used combinatorial domain mutations and most known AbdA developmental functions as biological readouts to investigate how protein domains collectively shape protein activity. The results uncover redundancy, interactivity, and multifunctionality of protein domains as salient features underlying overall AbdA protein activity, providing means to apprehend functional diversity and accounting for the robustness of Hox-controlled developmental programs. Importantly, the results highlight context-dependency in protein domain usage and interaction, allowing major modifications in domains to be tolerated without general functional loss. The non-pleoitropic effect of domain mutation suggests that protein modification may contribute more broadly to molecular changes underlying morphological diversification during evolution, so far thought to rely largely on modification in gene cis-regulatory sequences
IL-17A Expression Is Localised to Both Mononuclear and Polymorphonuclear Synovial Cell Infiltrates
This study examines the expression of IL-17A-secreting cells within the inflamed synovium and the relationship to in vivo joint hypoxia measurements.IL-17A expression was quantified in synovial tissue (ST), serum and synovial fluid (SF) by immunohistochemistry and MSD-plex assays. IL-6 SF and serum levels were measured by MSD-plex assays. Dual immunofluorescence for IL-17A was quantified in ST CD15+ cells (neutrophils), Tryptase+ (mast cells) and CD4+ (T cells). Synovial tissue oxygen (tpO(2)) levels were measured under direct visualisation at arthroscopy. Synovial infiltration was assessed using immunohistochemistry for cell specific markers. Peripheral blood mononuclear and polymorphonuclear cells were isolated and exposed to normoxic or 3% hypoxic conditions. IL-17A and IL-6 were quantified as above in culture supernatants.IL-17A expression was localised to mononuclear and polymorphonuclear (PMN) cells in inflamed ST. Dual immunoflourescent staining co-localised IL-17A expression with CD15+ neutrophils Tryptase+ mast cells and CD4+T cells. % IL-17A positivity was highest on CD15+ neutrophils, followed by mast cells and then CD4+T-cells. The number of IL-17A-secreting PMN cells significantly correlated with sublining CD68 expression (r = 0.618, p<0.01). IL-17A SF levels correlated with IL-6 SF levels (r = 0.675, p<0.01). Patients categorized according to tp0(2)< or >20 mmHg, showed those with low tp0(2)<20 mmHg had significantly higher IL-17A+ mononuclear cells with no difference observed for PMNs. Exposure of mononuclear and polymorphonuclear cells to 3% hypoxia, significantly induced IL-6 in mononuclear cells, but had no effect on IL-17A expression in mononuclear and polymorphonuclear cells.This study demonstrates IL-17A expression is localised to several immune cell subtypes within the inflamed synovial tissue, further supporting the concept that IL-17A is a key mediator in inflammatory arthritis. The association of hypoxia with Il-17A expression appears to be indirect, probably through hypoxia-induced pro-inflammatory pathways and leukocyte influx within the joint microenvironment
Rheumatoid arthritis: pathological mechanisms and modern pharmacologic therapies.
Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease that primarily affects the lining of the synovial joints and is associated with progressive disability, premature death, and socioeconomic burdens. A better understanding of how the pathological mechanisms drive the deterioration of RA progress in individuals is urgently required in order to develop therapies that will effectively treat patients at each stage of the disease progress. Here we dissect the etiology and pathology at specific stages: (i) triggering, (ii) maturation, (iii) targeting, and (iv) fulminant stage, concomitant with hyperplastic synovium, cartilage damage, bone erosion, and systemic consequences. Modern pharmacologic therapies (including conventional, biological, and novel potential small molecule disease-modifying anti-rheumatic drugs) remain the mainstay of RA treatment and there has been significant progress toward achieving disease remission without joint deformity. Despite this, a significant proportion of RA patients do not effectively respond to the current therapies and thus new drugs are urgently required. This review discusses recent advances of our understanding of RA pathogenesis, disease modifying drugs, and provides perspectives on next generation therapeutics for RA
Inhomogeneity of immune cell composition in the synovial sublining: linear mixed modelling indicates differences in distribution and spatial decline of CD68+ macrophages in osteoarthritis and rheumatoid arthritis
- …