Presentation_1_Wnts Promote Synaptic Assembly Through T-Cell Specific Transcription Factors in Caenorhabditis elegans.PDF

<p>Synapses are specialized neuronal connections essential for neuronal function. Defects in synaptic assembly or maintenance usually lead to various neurological disorders. Synaptic assembly is regulated by secreted molecules such as Wnts. Wnts are a large family of conserved glycosylated signaling molecules involved in many aspects of neural development and maintenance. However, the molecular mechanisms by which Wnts regulate synaptic assembly remain elusive due to the large number of ligands/receptors, the diversity of signaling cascades and the complexity of the nervous system. In this study, through genetic manipulation, we uncover that C. elegans Wnt-2 (CWN-2) is required for synaptic development. The CWN-2 signal is required during both embryonic and postembryonic development, in the nervous system and intestine, for promoting synaptic assembly. Furthermore, we provide genetic evidence for CWN-2 promoting synaptogenesis through the Frizzled receptor (FZD) CFZ-2, the Dishevelled (DVL) DSH-2, the β-catenin SYS-1 and the only T-cell specific transcription factor POP-1/TCF. Importantly, it is the first time to report the requirement of a TCF for presynaptic assembly. These findings expand our understanding of the synaptogenic mechanisms and may provide therapeutic insights into Wnt-related neurological disorders.</p

ABC294640 decreases monocyte/macrophage infiltration after liver transplantation.

<p>Livers were harvested at 18 h after transplantation (TX) or sham-operation (Sham). ED1 was detected immunohistochemically (<b>A–C</b>). ED1-positive cells were counted in a blinded manner in 10 randomly selected fields using a 40x objective lens (<b>D</b>). ED2 and actin in liver tissue were detected by immunoblotting. Representative blots and quantification by densitometry are shown in <b>E</b>. ABC, ABC294640; a, p<0.05 vs sham; b, p<0.05 vs the TX group (n = 4 per group).</p

ABC294640 blunts TLR4 and ICAM-1 upregulation and NF-κB activation after liver transplantation.

<p>Livers were harvested at 18 h after transplantation (TX) or sham-operation (Sham). Levels of TLR4, NF-κB p65 subunit, phosphorylated NF-κB p65 subunit (p-NF-κB p65), intracellular adhesion molecule-1 (ICAM-1) and actin were determined by immunoblotting. Representative blot images are shown in <b>A</b>. Images were quantified by densitometry (<b>B–D</b>). ABC, ABC294640; a, p<0.05 vs sham; b, p<0.05 vs the TX group (n = 4 per group).</p

ABC294640 attenuates necrosis and apoptosis after liver transplantation.

<p>Livers were harvested at 18 h after transplantation (TX) or sham-operation (Sham). Liver slices were stained with H&E for assessment of necrosis. Representative images are shown in <b>A to C</b>. Arrows identify the necrotic areas. The bar is 100 µm. Necrotic areas were quantified by image analysis of 10 randomly selected fields (<b>D</b>). Apoptosis was assessed by TUNEL staining, and TUNEL-positive cells were counted in 10 randomly selected fields (<b>E</b>). Cleaved caspase-3 and actin were detected by immunoblotting (<b>F</b>). ABC, ABC294640; a, p<0.05 vs sham; b, p<0.05 vs the TX group (n = 4 per group).</p

ABC294640 blunts TNFα, IL-1β and CXCL-10 mRNA increases after liver transplantation.

<p>Livers were harvested at 18 h after transplantation (TX) or sham-operation (Sham). Tumor necrosis factor-α (TNFα, <b>A</b>), interleukin-1β (IL-1β, <b>B</b>) and C-X-C motif chemokine 10 (CXCL-10, C) mRNAs in liver tissue were measured by quantitative real-time PCR. A.U., arbitrary units. ABC, ABC294640; a, p<0.05 vs sham; b, p<0.05 vs the TX group (n = 4 per group).</p

Real-Time PCR Primers.

<p>IL-1β, interleukin-1β; TNF-α, tumor necrosis factor; CXCL-10, C-X-C motif chemokine 10; HPRT, hypoxanthine phospho-ribosyl-transferase.</p

ABC294640 blunts CD4+ T Cell Infiltration and IFNγ Production after liver transplantation.

<p>Livers were harvested at 18 h after transplantation (TX) or sham-operation (Sham). CD4, IFNγ and actin were determined by immunoblotting and representative images are shown in <b>A</b>. CD4 and IFNγ images were quantified by densitometry (<b>B and C</b>). ABC, ABC294640; a, p<0.05 vs sham; b, p<0.05 vs the TX group (n = 4 per group).</p

ABC294640 decreases S1P production after liver transplantation.

<p>Livers were harvested at 18 h after transplantation (TX) or sham-operation (Sham). Sphingosine-1-phosphate (S1P) in liver tissues was quantified by ELISA. ABC, ABC294640; a, p<0.05 vs sham; b, p<0.05 vs the TX group (n = 4 per group).</p

Distribution of differentially expressed lncRNAs and mRNAs on chromosomess.

<p>Distribution of differentially expressed lncRNAs and mRNAs on chromosomess.</p

Analysis of DE-lincRNAs and neighboring protein-coding genes (distance < 300 kb).

<p>Analysis of DE-lincRNAs and neighboring protein-coding genes (distance < 300 kb).</p