Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 5 de 5
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Am J Physiol Renal Physiol ; 315(4): F812-F823, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-28468965

RESUMO

The urinary tract is usually culture negative despite its close proximity to microbial flora. The precise mechanism by which the kidneys and urinary tract defends against infection is not well understood. The initial kidney cells to encounter ascending pathogens are the collecting tubule cells that consist of principal cells (PCs) that express aquaporin 2 (AQP2) and intercalated cells (ICs) that express vacuolar H+-ATPase (V-ATPase, B1 subunit). We have previously shown that ICs are involved with the human renal innate immune defense. Here we generated two reporter mice, VATPase B1-cre+tdT+ mice to fluorescently label ICs and AQP2-cre+tdT+ mice to fluorescently label PCs, and then performed flow sorting to enrich PCs and ICs for analysis. Isolated ICs and PCs along with proximal tubular cells were used to measure antimicrobial peptide (AMP) mRNA expression. ICs and PCs were significantly enriched for AMPs. Isolated ICs responded to uropathogenic Escherichia coli (UPEC) challenge in vitro and had higher RNase4 gene expression than control while both ICs and PCs responded to UPEC challenge in vivo by upregulating Defb1 mRNA expression. To our knowledge, this is the first report of isolating murine collecting tubule cells and performing targeted analysis for multiple classes of AMPs.


Assuntos
Aquaporina 2/imunologia , Células Epiteliais/metabolismo , Túbulos Renais Coletores/imunologia , Reação em Cadeia da Polimerase , Animais , Aquaporina 2/genética , Imunidade Inata/imunologia , Rim/imunologia , Rim/metabolismo , Camundongos Transgênicos , Reação em Cadeia da Polimerase/métodos , Regulação para Cima/imunologia , ATPases Vacuolares Próton-Translocadoras/imunologia , ATPases Vacuolares Próton-Translocadoras/metabolismo
3.
Biomarkers ; 15(5): 424-35, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20491521

RESUMO

Currently there are no biomarkers for detecting collecting duct damage in man. Antibodies to several collecting duct-specific antigens exist but sandwich assays have been difficult to establish due to the need for two different antibodies to the same protein. We hypothesized that a collecting duct-specific lectin could be used in combination with a collecting duct-specific antibody to negate the need for two different antibodies. The collecting duct specificity of selected antibodies (NiCa II 13C2, Pap XI 3C7, HuPaP VII 2B11 and aquaporin 2), was verified by immunohistochemistry. Aquaporin 2 and Pap XI 3C7 were used successfully in setting up assays with the lectin Dolichos biflorus, using the Meso Scale Discovery (MSD) platform. Antigen expression was highest in the papillae of rat and human kidney (corresponding to the greatest density of collecting ducts) and was also present in normal urine. We propose that further qualification and validation would lead to an assay for detecting collecting duct damage in man.


Assuntos
Anticorpos/análise , Biomarcadores/análise , Imunoensaio/métodos , Túbulos Renais Coletores/imunologia , Lectinas de Plantas/imunologia , Animais , Antígenos/urina , Aquaporina 2/imunologia , Etilaminas , Humanos , Imuno-Histoquímica , Rim/imunologia , Rim/metabolismo , Necrose Papilar Renal/induzido quimicamente , Necrose Papilar Renal/imunologia , Necrose Papilar Renal/urina , Masculino , Ratos , Ratos Wistar
4.
Kidney Int ; 72(6): 731-5, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17597699

RESUMO

Immunocytochemistry performed on paraffin or cryosections is often hampered by poor morphology. Epoxy sections, in contrast, generally retain well-preserved tissue architecture. Immunocytochemistry, however, on epoxy-embedded sections is difficult due in part to the plastic itself and to the fixation conditions. Here, we present a technique for visualization of membrane proteins by immunocytochemistry on epoxy sections of kidneys fixed with glutaraldehyde without or with osmium post-fixation. Semithin sections were obtained from Epon 812-embedded mouse and rat kidney blocks. Before immunoperoxidase or immunofluorescence labeling, the sections were etched with the epoxy solvent, methanolic potassium hydroxide, followed by antigen retrieval using microwave heating. The sections were then treated with the primary antibody followed by secondary antibodies as usual. The distribution and expression patterns of a variety of membrane proteins, such as aquaporin (AQP)-1, AQP-2, and megalin, were identical to those observed by traditional immunocytochemical procedures on paraffin or cryosections. The advantages of our novel method include not only enhanced morphological quality but also the feasibility for investigators to visualize antigens of interest using archival specimens in Epon blocks.


Assuntos
Resinas Epóxi , Imunofluorescência , Técnicas Imunoenzimáticas , Rim/patologia , Proteínas de Membrana/metabolismo , Fixação de Tecidos/métodos , Animais , Anticorpos , Aquaporina 1/imunologia , Aquaporina 1/metabolismo , Aquaporina 2/imunologia , Aquaporina 2/metabolismo , Fixadores , Glutaral , Rim/metabolismo , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/imunologia , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Masculino , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Microtomia , Osmio , Ratos , Ratos Wistar , Simportadores de Cloreto de Sódio/imunologia , Simportadores de Cloreto de Sódio/metabolismo , ATPases Vacuolares Próton-Translocadoras/imunologia , ATPases Vacuolares Próton-Translocadoras/metabolismo
5.
Laryngoscope ; 117(4): 695-8, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17415141

RESUMO

OBJECTIVE: To localize aquaporin (AQP)2, vasopressin type 2 receptor (V2-R), and transient receptor potential channel vanilloid subfamily 1, 4 (TRPV1, TRPV4) in the human endolymphatic sac (ES). METHODS: Three samples of human ES were sampled during the removal of vestibular schwannoma by way of the translabyrinthine approach. The samples were immediately fixed in 4% paraformaldehyde and embedded in OCT compound; immunohistochemistry was performed with AQP2, V2-R, TRPV1, and TRPV4 polyclonal antibodies. RESULTS: AQP2, V2-R, TRPV1, and TRPV4 proteins were detected in the epithelial layer of the ES but were not observed in connective tissue around the ES. TRPV1 was also expressed in blood vascular endothelial cells of the connective tissue of ES. CONCLUSIONS: AQP2, V2-R, and TRPV4 were expressed in the luminal epithelium of human ES. The same characteristic distribution of water and ion channels is seen in the kidney, where a significant amount of fluid is filtrated and resorbed. ES probably plays an active role in the homeostasis of the endolymph.


Assuntos
Aquaporina 2/genética , Aquaporina 2/metabolismo , Saco Endolinfático/metabolismo , Neuroma Acústico/genética , Neuroma Acústico/metabolismo , Receptores de Vasopressinas/genética , Receptores de Vasopressinas/metabolismo , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Anticorpos/imunologia , Aquaporina 2/imunologia , Saco Endolinfático/imunologia , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Humanos , Imuno-Histoquímica , Neuroma Acústico/imunologia , Pressão Osmótica , Receptores de Vasopressinas/imunologia , Canais de Cátion TRPV/imunologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...