RESUMO
Fibrosis is the accumulation of extracellular matrix proteins and is a common end pathway in many chronic diseases. To identify novel mediators of fibrosis we used transcript profiling in a mouse model of kidney fibrosis, the COL4A3 knockout (alport) mouse. One gene that we found up-regulated in fibrotic kidney was GLIPR-2, also known as GAPR-1 and C9orf19, a member of the plant pathogenesis-related proteins family 1. We have found that GLIPR-2 protein expression is significantly increased in fibrotic kidney compared to healthy controls. Examination of the expression pattern of GLIPR-2 indicated that the protein is selectively expressed in epithelial cells. Co-staining with antibodies for alpha-smooth muscle actin expression, a marker of myofibroblasts, showed that GLIPR-2 expressing cells are closely apposed to areas of strong alpha-smooth muscle actin expression. The origin of these myofibroblasts is not known, but in vitro studies have shown that GLIPR-2 can induce epithelial to mesenchymal transition (EMT) in a renal epithelial cell line. We propose that increased GLIPR-2 expression in kidney contributes to development of fibrosis by increasing the pool of activated fibroblasts, possibly through the induction of EMT.
Assuntos
Diferenciação Celular/fisiologia , Células do Tecido Conjuntivo/citologia , Células Epiteliais/citologia , Rim/metabolismo , Proteínas/metabolismo , Actinas/metabolismo , Animais , Autoantígenos/genética , Caderinas/metabolismo , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Linhagem Celular , Colágeno Tipo IV/genética , Células do Tecido Conjuntivo/efeitos dos fármacos , Células do Tecido Conjuntivo/metabolismo , Modelos Animais de Doenças , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Fibrose , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Rim/patologia , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Medula Renal/metabolismo , Túbulos Renais/metabolismo , Túbulos Renais/patologia , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas de Membrana/farmacologia , Camundongos , Camundongos Endogâmicos , Camundongos Knockout , Proteínas/genética , Transfecção , Vimentina/metabolismoRESUMO
The use of microarrays to monitor gene expression has become a standard research tool at both academic and industrial research institutions. Quality control of common printing defects during DNA deposition onto glass substrates is critical to maintaining data integrity and preventing the needless consumption of precious RNA, labeling reagents, and time. Here we demonstrate a nondestructive method for monitoring the quality of every spot on every chip of a microarray production run. We have identified many common manufacturing defects, while not perturbing the attachment of our oligonucleotide target to the substrate or altering further hybridization. This protocol is simple, fast, and inexpensive.