Rapid communication: Physical and genetic mapping of the Peroxisome Proliferator Activated Receptor γ (PPARγ) gene to porcine chromosome 13

Genus and Species. Sus scrofa. Locus Name. Peroxisome Proliferator Activated Receptor gamma (PPARγ). Source and Description of Primers. Primers were designed in exon 5 from a published porcine cDNA sequence (GenBank accession no. AJ006756). Forward primer: 5′ GAC ATG AAT TCC TTA ATG 3′; reverse primer: 5′ ACT TCA CAG CGA ACT CGA ACT T 3′

T Cell Specific Adapter Protein (TSAd) Interacts with Tec Kinase ITK to Promote CXCL12 Induced Migration of Human and Murine T Cells

The chemokine CXCL12/SDF-1α interacts with its G-protein coupled receptor CXCR4 to induce migration of lymphoid and endothelial cells. T cell specific adapter protein (TSAd) has been found to promote migration of Jurkat T cells through interaction with the G protein β subunit. However, the molecular mechanisms for how TSAd influences cellular migration have not been characterized in detail. We show that TSAd is required for tyrosine phosphorylation of the Lck substrate IL2-inducible T cell kinase (Itk). Presence of Itk Y511 was necessary to boost TSAd\u27s effect on CXCL12 induced migration of Jurkat T cells. In addition, TSAd\u27s ability to promote CXCL12-induced actin polymerization and migration of Jurkat T lymphocytes was dependent on the Itk-interaction site in the proline-rich region of TSAd. Furthermore, TSAd-deficient murine thymocytes failed to respond to CXCL12 with increased Itk phosphorylation, and displayed reduced actin polymerization and cell migration responses. We propose that TSAd, through its interaction with both Itk and Lck, primes Itk for Lck mediated phosphorylation and thereby regulates CXCL12 induced T cell migration and actin cytoskeleton rearrangements

The pathophysiological function of peroxisome proliferator-activated receptor-γ in lung-related diseases

Research into respiratory diseases has reached a critical stage and the introduction of novel therapies is essential in combating these debilitating conditions. With the discovery of the peroxisome proliferator-activated receptor and its involvement in inflammatory responses of cardiovascular disease and diabetes, attention has turned to lung diseases and whether knowledge of this receptor can be applied to therapy of the human airways. In this article, we explore the prospect of peroxisome proliferator-activated receptor-γ as a marker and treatment focal point of lung diseases such as asthma, chronic obstructive pulmonary disorder, lung cancer and cystic fibrosis. It is anticipated that peroxisome proliferator-activated receptor-γ ligands will provide not only useful mechanistic pathway information but also a possible new wave of therapies for sufferers of chronic respiratory diseases

Rapid communication: Physical and genetic mapping of the Peroxisome Proliferator Activated Receptor γ (PPARγ) gene to porcine chromosome 13

Genus and Species. Sus scrofa. Locus Name. Peroxisome Proliferator Activated Receptor gamma (PPARγ). Source and Description of Primers. Primers were designed in exon 5 from a published porcine cDNA sequence (GenBank accession no. AJ006756). Forward primer: 5′ GAC ATG AAT TCC TTA ATG 3′; reverse primer: 5′ ACT TCA CAG CGA ACT CGA ACT T 3′.This is an article from Journal of Animal Science 78 (2000): 1391, doi:/2000.7851391x. Posted with permission.</p

CXCL12 mediated Itk tyrosine phosphorylation is dependent on TSAd expression in T cells.

<p>(A) Jurkat T cells transiently transfected with plasmids encoding the indicated proteins were kept unstimulated (−) or stimulated (+) with 500 ng/ml CXCL12 for 2 min and then lysed. Itk was immunoprecipitated with anti-myc antibody and immunoblotted with anti-phosphotyrosine antibody. The blot was stripped and reprobed with anti-Itk antibody to ensure equal immunoprecipitation. The corresponding lysates were probed with the indicated antibodies to verify expression from the transfected plasmids. (B) Thymocytes from <i>SH2D2A</i>^+/+ or <i>SH2D2A</i>^−/− mice were stimulated with 100 ng/ml CXCL12 for 2 minutes (+), lysed and subjected to immunoprecipitation with anti-Itk antibody. Immunoprecipitates were subjected to immunoblotting with anti-phosphotyrosine antibody to detect tyrosine phosphorylated Itk. The blot was stripped and reprobed with anti-Itk antibody to ensure equal immunoprecipitation efficiency, and with anti-Lck, anti-Zap70 and anti-mTSAd (murine TSAd) antibodies to detect co-immunoprecipitation of these proteins with Itk. Lysates were probed with anti-mTSAd, anti-Lck and anti-Zap70 antibodies to verify their expression.</p

CXCL12 mediated actin polymerization and cellular migration is impaired in murine TSAd-deficient thymocytes.

<p>(A) Thymocytes from <i>SH2D2A</i>^+/+ or <i>SH2D2A</i>^−/− mice were left unstimulated (0) or stimulated with 10, 100, 500 or 1000 ng/ml CXCL12 for 15 seconds and stained for F actin. The F-actin content was assigned as mean fluorescence intensity value of FITC-phalloidin stained cells. Data shown are F-actin content after 15 seconds of stimulation with the indicated CXCL12 concentrations relative to F-actin content in unstimulated cells. The graph represents the mean value from at least four independent experiments, +- SEM, *  =  P<0,05. (B) Migration of thymocytes from <i>SH2D2A</i>^+/+ or <i>SH2D2A</i>^−/− mice in response to medium alone or indicated amounts of CXCL12 was assessed using a transwell migration assay. Migration index (MI) was calculated as the number of cells migrating towards CXCL12 relative to the number of cells migrating towards medium only. The graph shows mean MI values +/− SEM of at least four independent assays. The Western blot verifies expression of murine TSAd (mTSAd). (C) Chemokinesis index is calculated as spontaneous migration of either <i>SH2D2A^+/+</i> or <i>SH2D2A^−/−</i> thymocytes relative to that observed for <i>SH2D2A</i>^+/+ thymocytes in each independent experiment presented in B. The graph shows mean chemokinesis index values +/− SEM of eight independent assays. (D) <i>SH2D2A</i>^+/+and <i>SH2D2A</i>^−/− thymocytes prior to (top panels) or after (lower panels) CXCL12 induced migration were stained with fluorescently labeled anti-CD4 and CD8 antibodies and analyzed by flow cytometry. To achieve sufficient cells for fluorescently labeling after migration, 4×10⁶ thymocytes were allowed to migrate for 4 hours prior to staining. (E) Thymocytes from <i>SH2D2A</i>^+/+ and <i>SH2D2A</i>^−/− mice were analyzed for CXCR4 expression by flow cytometry. Left and middle panel: grey areas - control antibody staining; open areas - CXCR4 staining, right panel: grey areas - CXCR4 staining of <i>SH2D2A</i>^+/+ cells, open areas – CXCR4 staining of <i>SH2D2A</i>^−/− cells. (F) Migration of thymocytes from <i>SH2D2A</i>^+/+ or <i>SH2D2A</i>^−/− mice in response to medium alone or indicated amounts of CCL19 was assessed using a transwell migration assay. MI was calculated as in panel 5B. The graph shows mean MI values +/− SEM of at least three independent assays.</p