RESUMEN
Clara cell secretory protein (CCSP) is the most abundant secreted protein within airways of the lung. Moreover, CCSP levels are modulated in human lung disease, supporting a potentially important role for CCSP and/or Clara cells in lung homeostasis. However, in vivo roles for CCSP remain elusive. A popular hypothesis is that CCSP is a regulator of the inflammatory response. The purpose of this review is to provide an overview of the phenotype of CCSP null mice and relate this phenotype to proposed functions for the protein. Phenotypic analysis of mice homozygous for the CCSP-1 null allele of the CCSP gene (CCSP-/-1) revealed susceptibility to inhaled oxidant gases. Sensitivity of CCSP-/-1 mice to inhaled ozone is unrelated to alterations in antioxidant defenses, but is associated with increased cellular injury. Additional studies investigating inflammatory control in CCSP deficient mice found no differences between wild-type and CCSP-/-1 mice in their inflammatory response to low-dose inhaled endotoxin exposure, arguing against a role for CCSP in regulation of pulmonary inflammation. The findings among CCSP-/-1 mice of ultrastructural alterations to Clara cell secretory apparatus, with associated changes in airway lining fluid protein composition, demonstrate that the CCSP-/-1 genotype results in more complex changes to airways than CCSP deficiency per se. It can be concluded that CCSP does not regulate endotoxin-induced pulmonary inflammation. Moreover, CCSP-/-1 mice represent a valuable tool for probing functional roles for Clara cells in regulation of airway lining fluid composition and lung pollutant susceptibility.
Asunto(s)
Pulmón/metabolismo , Neumonía/genética , Proteínas/genética , Mucosa Respiratoria/metabolismo , Uteroglobina , Contaminantes Atmosféricos/efectos adversos , Animales , Humanos , Pulmón/patología , Pulmón/fisiopatología , Ratones , Ratones Noqueados/genética , Ratones Noqueados/metabolismo , Fenotipo , Neumonía/metabolismo , Proteínas/metabolismoRESUMEN
Clara cell secretory protein (CCSP) is one of the most abundant proteins present in airway lining fluid of mammals. In an effort to elucidate the function of CCSP, we established CCSP-null [CCSP(-/-)] mice and demonstrated altered sensitivity to various environmental agents including oxidant pollutants and microorganisms. Although CCSP deficiency itself may be central to the observed changes in environmental susceptibility, altered lung gene expression associated with CCSP deficiency may contribute to the observed phenotype. To determine whether CCSP deficiency results in altered lung gene expression, high-density cDNA microarrays were used to profile gene expression in the total lung RNA of wild-type and CCSP(-/-) mice. Genes that were differentially expressed between wild-type and CCSP(-/-) mice included a previously non-annotated expressed sequence tag (EST W82219) and immunoglobulin A (IgA), both of which were elevated with CCSP deficiency. mRNA expression of EST W82219 and IgA was localized in the lungs of wild-type and CCSP(-/-) mice to airway Clara cells and peribronchial lymphoid tissues, respectively. We conclude that CCSP deficiency is associated with 1) altered gene expression in Clara cells of the conducting airway epithelium and 2) alterations to peribronchial B lymphocytes. These findings identify new roles for Clara cells and their secretions in airway homeostasis.
Asunto(s)
Proteínas/genética , Mucosa Respiratoria/citología , Mucosa Respiratoria/inmunología , Uteroglobina , Animales , Bronquios/citología , Bronquios/inmunología , Bronquios/metabolismo , Expresión Génica/inmunología , Hiperoxia/inmunología , Hiperoxia/metabolismo , Inmunoglobulina A/genética , Inmunoglobulina A/inmunología , Ratones , Ratones Endogámicos , Ratones Noqueados , Oxidación-Reducción , Fenotipo , ARN Mensajero/análisis , Mucosa Respiratoria/metabolismoRESUMEN
Clara cell secretory protein (CCSP), also known as uteroglobin (Ug), is a 16-kDa homodimeric protein of unknown function. Within rodent species, CCSP is expressed predominantly by nonciliated Clara cells that line conducting airways of the lung. To investigate in vivo functions for CCSP, we established mice homozygous for a null allele of the CCSP gene (CCSP-/-). We previously showed no overt phenotypic consequences associated with CCSP deficiency when CCSP-/- mice are maintained in the absence of environmental stress. However, CCSP-/- mice show an oxidant-sensitive phenotype that cannot be attributed to alterations in the inflammatory response when challenged by inhaled oxidant gases. The current study was undertaken to determine whether CCSP deficiency results in pathological changes to the kidney. This study was prompted by the recent description of severe systemic disease and kidney fibrosis/dysfunction in an independent line of CCSP-deficient mice, termed Ug-/- (Zhang et al, Science 276:1408-1412, 1997). CCSP-/- mice show normal growth and reproductive performance when maintained in two independent genetic backgrounds, inbred 129 and congenic C57BL/6. Strain 129 CCSP-/- mice have normal kidney function, as assessed by urinary glucose, lactate dehydrogenase, and glomerular filtration rate; they show no kidney fibrosis or abnormalities in fibronectin accumulation and no histological abnormalities in proximal convoluted tubules or glomeruli at either light or electron microscopic levels. CCSP deficiency is associated with mild proteinurea involving a modest increase in mouse major urinary protein-1. We conclude that CCSP (Ug) deficiency, per se, is not the cause of severe renal pathology and systemic disease reported for Ug-/- mice.
Asunto(s)
Fibronectinas/metabolismo , Riñón/metabolismo , Proteínas/metabolismo , Uteroglobina/deficiencia , Animales , Western Blotting , Técnica del Anticuerpo Fluorescente , Tasa de Filtración Glomerular , Glucosuria/orina , Riñón/fisiopatología , Enfermedades Renales/metabolismo , Pruebas de Función Renal , L-Lactato Deshidrogenasa/orina , RatonesRESUMEN
Clara cell secretory protein (CCSP) is a transport protein for lipophilic substances in bronchio-alveolar fluid, plasma, and uterine secretion. It acts as a carrier for steroid hormones and polychlorinated biphenyl metabolites. Previously, the existence of receptors for uptake of CCSP.ligand complexes into the renal proximal tubules had been suggested. Using surface plasmon resonance analysis, we demonstrate that CCSP binds to cubilin, a peripheral membrane protein on the surface of proximal tubular cells. Binding to cubilin results in uptake and lysosomal degradation of CCSP in cultured cells. Surprisingly, internalization of CCSP is blocked not only by cubilin antagonists but also by antibodies directed against megalin, an endocytic receptor that does not bind CCSP but associates with cubilin. Consistent with a role of both receptors in renal uptake of CCSP in vivo, patients deficient for cubilin or mice lacking megalin exhibit a defect in tubular uptake of the protein and excrete CCSP into the urine. These findings identify a cellular pathway consisting of a CCSP-binding protein (cubilin) and an endocytic coreceptor (megalin) responsible for tissue-specific uptake of CCSP and associated ligands.