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1.
J Nephrol ; 20(3): 357-63, 2007.
Article in English | MEDLINE | ID: mdl-17557270

ABSTRACT

BACKGROUND: An increased number of major histocompatibility complex (MHC) class II-positive cells (OX-6+ cells) were observed in the glomerulus and periglomerular interstitium during the course of anti-glomerular basement membrane (anti-GBM) crescentic glomerulonephritis (GN) in WKY rats. This study aimed to demonstrate that periglomerular OX-6+ cells are dendritic cells (DCs) and to clarify their roles in the pathogenesis of this GN. METHODS: Kidney sections were stained with the OX-6 and the rat DC marker OX-62 by immunohistochemistry, and periglomerular OX-6+ cells were observed by immunoelectron microscopy. Renal mRNA expression for CXCL12 was examined by reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization, and that for IL-1beta was examined by in situ hybridization. RESULTS: Immunohistochemistry revealed that most periglomerular OX-6+ cells in this GN were ED-1-negative. OX-62+ cells were observed sparsely in normal kidney interstitium, and considerably more frequently in periglomerular interstitium in this GN. Immunoelectron microscopy confirmed the periglomerular OX-6+ED-1- cells had DC morphology. The increased expression of CXCL12 mRNA in the diseased glomerulus was shown by RT-PCR. By in situ hybridization, CXCL12 mRNA-expressing glomerular cells were the parietal and visceral epithelial cells, which were close to the site of periglomerular OX-6+ cell localization. The intense expression of IL-1beta mRNA by periglomerular cells was demonstrated by in situ hybridization. CONCLUSIONS: The periglomerular distribution of OX-6+ED-1- DCs was demonstrated in anti-GBM crescentic GN in WKY rats. These DCs might be accumulated in periglomerular interstitium by CXCL12, and play a role in the initiation and progression of this GN by producing IL-1beta.


Subject(s)
Dendritic Cells , Glomerulonephritis/pathology , Animals , Kidney Glomerulus , Rats , Rats, Inbred WKY
2.
Am J Physiol Regul Integr Comp Physiol ; 292(1): R644-51, 2007 Jan.
Article in English | MEDLINE | ID: mdl-16990491

ABSTRACT

Hagfish (Eptatretus burgeri) are agnathous and are the earliest vertebrates still in existence. Pavement cells adjacent to the mitochondria-rich cells show orthogonal arrays of particles (OAPs) in the gill of hagfish, a known ultrastructural morphology of aquaporin (AQP) in mammalian freeze-replica studies, suggesting that an AQP homolog exists in pavement cells. We therefore cloned water channels from hagfish gill and examined their molecular characteristics. The cloned AQP [E. burgeri AQP4 (EbAQP4)] encodes 288 amino acids, including two NPA motifs and six transmembrane regions. The deduced amino acid sequence of EbAQP4 showed high homology to mammalian and avian AQP4 (rat, 44%; quail, 43%) and clustered with AQP4 subsets by the molecular phylogenetic tree. The osmotic water permeability of Xenopus oocytes injected with EbAQP4 cRNA increased eightfold compared with water-injected controls and was not reversibly inhibited by 0.3 mM HgCl(2). EbAQP4 mRNA expression in the gill was demonstrated by the RNase protection assay; antibody raised against the COOH terminus of EbAQP4 also detected (by Western blot analysis) a major approximately 31-kDa band in the gill. Immunohistochemistry and immunoelectron microscopy showed EbAQP4 localized along the basolateral membranes of gill pavement cells. In freeze-replica studies, OAPs were detected on the protoplasmic face of the split membrane comprising particles 5-6 nm long on the basolateral side of the pavement cells. These observations suggest that EbAQP4 is an ancestral water channel of mammalian AQP4 and plays a role in basolateral water transport in the gill pavement cells.


Subject(s)
Hagfishes/metabolism , Amino Acid Sequence , Animals , Biological Transport, Active/physiology , Blotting, Western , Cell Membrane/metabolism , DNA, Complementary/biosynthesis , DNA, Complementary/genetics , Freeze Fracturing , Gills/metabolism , Immunohistochemistry , Mercury/pharmacology , Microscopy, Electron , Molecular Sequence Data , Multigene Family/genetics , Oocytes/metabolism , Permeability , Phylogeny , RNA/biosynthesis , RNA/genetics , Reverse Transcriptase Polymerase Chain Reaction , Water/metabolism , Xenopus
3.
Kidney Int ; 65(4): 1280-9, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15086467

ABSTRACT

BACKGROUND: Investigated were effects of overexpression of interleukin-10 (IL-10) on the outcome and progression of crescentic glomerulonephritis in Wistar-Kyoto (WKY) rats. METHODS: Rats were singly or simultaneously injected with antiglomerular basement membrane (a-GBM) antibody and adenoviral vector encoding rat IL-10 (Ad-rIL-10) or LacZ (Ad-LacZ) (3 x 1010 pfu/rat) intravenously, and were sacrificed at day 7. Their kidneys and other organs were isolated and examined by histology and immunohistochemistry. The In vivo expression of IL-10 mRNA in the liver of Ad-rIL-10-injected rats was confirmed by both reverse transcription-polymerase chain reaction (RT-PCR) and ribonuclease protection assay analysis and its translated protein was measured in the serum by enzyme-linked immunosorbent assay (ELISA). RESULTS: The exogenous IL-10 mRNA was strongly expressed in the liver in a dose-dependent manner and was intense at days 4 and 7 but was less intense at day 14. Ad-rIL-10 treatment significantly reduced the incidence of glomerular crescent formation from 67%+/- 1.9% in a-GBM antibody-treated group or 69.8%+/- 1.9% in a-GBM antibody + Ad-LacZ-treated group to 21.6%+/- 1.8% (P < 0.001), the glomerular infiltration of macrophages from 35.7 +/- 6.3 cell s/gcs (a-GBM antibody) or 37.6 +/- 8.6 cells/gcs (both a-GBM antibody + Ad-LacZ) to 17.9 +/- 5.5 cells/gcs (P < 0.001), that of major histocompatibility complex (MHC) class II-positive cells from 14.4 +/- 5.3 cells/gcs (a-GBM antibody) or 15 +/- 4.6 cells/gcs (a-GBM antibody + Ad-LacZ) to 5.7 +/- 2.3 cells/gcs (P < 0.0001) at day 7, the glomerular and immune tissue expression of IL-1beta mRNA, as well as the proteinuria from 159.0 +/- 22.7 mg/24 hours (a-GBM antibody) or 166 +/- 28 mg/24 hours (a-GBM antibody + Ad-LacZ) to 42.2 +/- 35.2 mg/24 hours (P < 0.01) at day 7. The serum creatinine and blood urea nitrogen levels were also reduced from 2.8 +/- 0.1 mg/dL (a-GBM antibody) or 2.8 +/- 0.1 mg/dL (a-GBM antibody + Ad-LacZ) to 1.0 +/- 0.1 mg/dL (P < 0.001) and from 63.2 +/- 8.9 mg/dL (a-GBM antibody) or 61.3 +/- 5.2 mg/dL (a-GBM antibody + Ad-LacZ) to 27.0 +/- 4.5 mg/dL (P < 0.001), respectively. However, the glomerular accumulation of CD8+ T cells was unaffected: 5.4 +/- 1.1 cells/gcs (a-GBM antibody + Ad-rIL-10), 5.9 +/- 1.5 cells/gcs (a-GBM antibody), and 5.8 +/- 1.1 cells/gcs (a-GBM antibody + Ad-LacZ) (P= NS). CONCLUSION: IL-10 gene transfer significantly attenuated the glomerular lesions and injury in the anti-GBM crescentic glomerulonephritis of WKY rats.


Subject(s)
Anti-Glomerular Basement Membrane Disease/metabolism , Anti-Glomerular Basement Membrane Disease/pathology , Gene Transfer Techniques , Interleukin-10/genetics , Adenoviridae/genetics , Animals , Blood Urea Nitrogen , Creatinine/blood , Disease Progression , Female , Gene Expression , Genetic Vectors , Immunohistochemistry , Interleukin-1/genetics , Kidney Glomerulus/metabolism , Lymph Nodes/metabolism , Proteinuria/urine , RNA, Messenger/antagonists & inhibitors , RNA, Messenger/metabolism , Rats , Rats, Inbred WKY , Spleen/metabolism
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