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132 117 P PACAP VIP PACAP VIP 1 2 2 1 1 2 1 PACAP VIP PACAP 8 C57BL/6 PACAP PAC1R VIP VPAC1 PAC1R pillar cell VPAC1 pillar cell PAC1R VPAC1 PACAP VIP P 1,2 2 2 1 2 2 1 2 5 AQP5 AQP5 AQP5 AQP5 9 Wistar AQP5 3 AQP5 AQP5 AQP5 P gustducin 2 116 gustducin G NSE 4 Hartley 4%PFA calbindin gustducin ABC gustducin TEM 0.5-1 μm ABC gustducin gustducin calbindin TEM gustducin P ZnT ZnT7 ZnT5 ZnT2 ZnT5 Se Zn ZnSO45 mg/100 g ZnT H1250M P 1 1 1 2 1 2 1 1 2 P APC Min/+ 1 1 1 2 1 2 HSC E HSC δ HSC HSCT6 δ HepG2 α HepG2 4 δ FACS TUNEL δ HSC δ β1 APC Min/+ Interstitial cells of Cajal; ICCICC ICC 7 APC Min/+ ICC ICCMPICCDMP cKit cKit ICCMP ICC ICC ICC
117 133 P DPEP 1 2 2 1 2 1 2 Dipeptidase1 DPEP1 dipeptideglutathioneleukotrieneD4 GPI DPEP1 HCC56 caveolin1 flotillin1 GPI CD59 DPEP1 GPI P Placenta specific miR-517a modulates gene expression in Jurkat cells Md Moksed Ali 1 , Xiaohui Song 1,2 , Osamu Ishibashi 1 , Kunio Kikuchi 1 , Tomoko Ishikawa 1 , Takami Takizawa 1 , Toshihiro Takizawa 1 1 Dept Mol Med & Anat, Nippon Medical School 2 Dept Pharm, Harbin Medical University [Objective] MicroRNAs miRNAs are noncoding RNAs that repress gene expression posttranscriptionally. Recently we have found that human placenta specific miRNAs are transferable to Jurkat cells human T cell lymphoblast like cell line via exosomes manuscript in preparation. In this study, we overexpressed placenta specific miR-517a in Jurkat cells and did microarray analysis MA to identify its target mRNAs. [Methods] MA was performed on Jurkat cells overexpressing miR-517a. Validation of genes downregulated by miR-517a was carried out by realtime PCR, Western blot, and 3'UTRluciferase reporter assay. [Results] We found that cyclic GMP dependent protein kinase 1 (PRKG1), one of 8 downregulated genes identified in MA, is a target of miR-517a in silico. MiR- 517a significantly downregulated the expression of PRKG1 at both the mRNA and protein levels. Furthermore, miR-517a significantly decreased luciferase reporter activity. [Conclusion] We identified that PRKG1 is indeed a target of miR-517a in Jurkat cells. These findings provide a mechanistic insight on the posttranscriptional regulation by miRNAs of placentaderived exosomes in T cells. P microRNA CpG 1 1,2 3 1 1 2 3 microRNA miRNA 22 RNA 19 miRNAs C19MC; miR-517a Luo et al. Biol Reprod 81: 717729, 2009 18 kb CG DNA C19MC NoguerDance et al. Hum Mol Genet 19: 35663582, 2010 BeWo JEG3 HTR8/SVneo C19MC miRNA PCR DNA PCR CG BeWoJEG3 C19MC HTR8/SVneo BeWo, JEG3 C19MC HTR8/SVneo P IgG IIb Fc FcRIIb 1 2 1 3 1 1 2 3 HPEC IgG HPEC IIb Fc FcRIIb IgG J Immunol 175: 2331, 2005 FcRIIb RAB GTPasesRAB1B RAB3D FcRIIb in vitro FcRIIb GFP FcRIIb pFCGR2B-GFP HUVEC FcRIIb RAB1BRAB3D siRNA FcRIIbsiRAB1B siRAB3D HUVEC pFCGR2B-GFP GFPFcRIIb GFPFcRIIb HUVEC siRNA GFPFcRIIb GFPFcRIIbHPEC RAB1B RAB3D FcRIIb P Agerelated accumulation of nonheme ferric and ferrous iron in mouse ovarian stroma visualized by sensitive iron histochemistries Yoshiya Asano Department of Neuroanatomy, Cell Biology and Histology, Hirosaki University Graduate School of Medicine Sensitive nonheme iron histochemistries, namely perfusionPerls and Turnbull method, were applied to study the distribution and agerelated accumulation of nonheme iron in mouse ovary. Microscopic studies revealed that nonheme ferric iron is distributed predominantly in stromal tissue, especially in macrophages. By contrast, the distribution of nonheme ferrous iron was restricted to a few ovoid macrophages. Aged ovaries exhibited remarkable nonheme iron accumulation in all stromal cells. Particularly, nonheme ferrous iron level was increased in stroma, suggestive of increased levels of redoxactive iron, which can promote oxidative stress. Moreover, intense localization of both nonheme ferric and ferrous iron was observed in aggregated large stromal cells that were then characterized as ceroid laden enlarged macrophages. Iron overload in adult mice resulted in nonheme iron deposition in the stroma and generation of enlarged macrophages, suggesting that iron accumulation induced morphological changes. Our data indicated that nonheme iron accumulation in stromal tissue may be related to aging of the ovary which due to the increasing of oxidative stress. P CD34 CD141 5 μm CD34 CD141 DAB CCD CD34 CD141 CD34 CD34
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192 117 Marson, Lesley Masahiro,
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