bromochloroacetic-acid and Polycystic-Kidney-Diseases

Topics: CD56 Antigen; Encephalocele; Fetus; Gene Expression Regulation, Developmental; Humans; Keratin-20; Keratin-7; Keratins; Liver; Polycystic Kidney Diseases; Polydactyly; Syndrome

Caroli's disease (congenital intrahepatic biliary dilatation) associated with congenital hepatic fibrosis is an autosomal recessive polycystic kidney disease. Recently, the polycystic kidney (PCK) rat, a spontaneous mutant derived from a colony of CRJ:CD rats with polycystic lesions in the liver and an autosomal recessive mode of inheritance, was reported. In the present study, the pathology of the hepatobiliary system and the biliary cell-kinetics were evaluated in fetuses (day 18 to 21 of gestation) and neonates and adults (1 day to 4 months after delivery) of PCK rats. CRJ:CD rats were used as a control. Multiple segmental and saccular dilatations of intrahepatic bile ducts were first observed in fetuses at 19 days of gestation. The dilatation spread throughout the liver and the degree of dilatation increased with aging. Gross and histological features characterizing ductal plate malformation were common in the intrahepatic bile ducts. Overgrowth of portal connective tissue was evident and progressive after delivery. These features were very similar to those of Caroli's disease with congenital hepatic fibrosis. Proliferative activity in the biliary epithelial cells was greater in PCK rats than controls during the development. In contrast, the biliary epithelial apoptosis was less extensive in PCK rats than the controls until 1 week after delivery, but greater after 3 weeks, suggesting that the remodeling defect in immature bile ducts associated with the imbalance of cell kinetics plays a role in the occurrence of intrahepatic biliary anomalies in PCK rats. The PCK rat could be a useful and promising animal model of Caroli's disease with congenital hepatic fibrosis.

Topics: Animals; Bile Ducts, Intrahepatic; Caroli Disease; Disease Models, Animal; Female; Immunohistochemistry; In Situ Nick-End Labeling; Intermediate Filament Proteins; Keratin-20; Keratins; Ki-67 Antigen; Kidney; Liver; Liver Cirrhosis; Male; Polycystic Kidney Diseases; Rats; Time Factors

The appearance of interstitial fibrosis in polycystic kidneys is emblematic of progressive disease. Matrix forming this scar tissue is derived from local renal cells in response to cystogenesis. We investigated the phenotype of collagen-producing cells in the cystic kidneys of DBA/2-pcy mice to better characterize the spectrum of interstitial cells associated with renal fibrogenesis.. The extent of interstitial fibrosis and the number of fibroblasts in cystic kidneys were first quantitated over time using computer-assisted image analysis. Subsequently, antisera to four cell protein markers were studied by coexpression immunohistochemistry during progression of fibrosis using confocal microscopy. The antisera included fibroblast-specific protein 1 (FSP1) for fibroblast phenotype, alpha-smooth muscle actin (alpha-SMA) for contractile phenotype, vimentin (VIM) for mesenchymal phenotype, and heat shock protein 47 (HSP47) for interstitial collagen-producing phenotype.. Interstitial fibrosis in cystic kidneys gradually increased throughout the 30-week observation period of our study. With progression of cystogenesis, most of the tubules in pcy mice either dilated or disappeared with time. FSP1+ fibroblasts were distributed sparsely throughout the renal interstitium of young pcy and wild-type mice. Their number increased in the widening fibrotic septa by 18 weeks of age and persisted through 30 weeks of the study interval. Some epithelia among remnant tubules trapped within fibrotic septa around adjacent cysts also acquired the phenotype of FSP1+, HSP47+ collagen-producing fibroblasts, suggesting a possible role for epithelial-mesenchymal transformation (EMT) in this process. Most FSP1+ fibroblasts were alpha-SMA-, but HSP47+, suggesting they were producing collagen proteins for the extracellular matrix. alpha-SMA+, FSP1-, HSP47+ or HSP47- cells were also observed, and the latter tended to distribute independently in a linear pattern, reminiscent of vasculature adjacent to forming cysts. VIM+ expression was not observed in alpha-SMA+ cells.. Many nonoverlapping as well as fewer overlapping populations of FSP1+ and alpha-SMA+ cells shared in the collagen expression associated with progressive fibrogenesis in pcy mice undergoing cystogenesis. Some FSP1+ fibroblasts are likely derived from tubular epithelium undergoing EMT, while alphaSMA+, VIM- cells probably represent vascular smooth muscle cells or pericytes surviving vessel attenuation during the chaos of fibrogenesis. Importantly, not all interstitial cells producing collagens are alpha-SMA+.

Topics: Actins; Animals; Biomarkers; Cadherins; Calcium-Binding Proteins; Fibroblasts; Fibrosis; Fluorescent Antibody Technique; Heat-Shock Proteins; HSP47 Heat-Shock Proteins; Keratins; Kidney; Male; Mice; Mice, Inbred DBA; Polycystic Kidney Diseases; S100 Calcium-Binding Protein A4; S100 Proteins

Renal cystic disease include heritable, developmental and acquired disorders. Morphological features were extensively studied mainly in cases of autosomal dominant polycystic and experimentally induced cystic disorders. We report the immunohistochemical (cytokeratin, epithelial membrane antigen, vimentin, Tamm-Horsfall protein, proliferating cell nuclear antigen) and lectin-binding (soybean agglutinin, Dolichos biflorus agglutinin) profile of cystic kidneys from 9 surgically removed and 21 autopsy cases. The primary renal diseases displayed great diversity. Beside polycystic kidney diseases we studied cysts associated to renal neoplasm, hemodialysis, nephrosis syndrome and chronic transplant rejection. Cystic epithelium demonstrated positive reactions with distal tubular markers (epithelial membrane antigen, cytokeratin) or collecting duct (soybean agglutinin, Dolichos biflorus agglutinin) and Henle loop markers (Tamm-Horsfall protein) but the latter in lesser extent. The large number of the vimentin positive cases are suggestive to dedifferentiation or cellular regeneration. The former might be underlined by the diffuse cytoplasmic or basolateral membrane staining of the epithelial membrane antigen in some cystic epithelial cells. Not the cystic epithelium but rather the neighbouring non-dilated tubular cells and interstitial cells presented proliferative activity which was most intense in areas where vimentin and variable nephron segment markers in the same tissue were expressed. Positive reaction of the type IV basement membrane collagen and the rate of the inflammation failed to show similar connection. This finding suggests the importance of the inflammatory cells in the development and/or expansion of the cysts.

Topics: Cell Differentiation; Cell Division; Collagen; Cytoplasm; Epithelium; Genes, Dominant; Glycine max; Graft Rejection; Humans; Immunohistochemistry; Keratins; Kidney Diseases, Cystic; Kidney Neoplasms; Kidney Tubules; Kidney Tubules, Collecting; Lectins; Loop of Henle; Mucin-1; Mucoproteins; Nephrotic Syndrome; Plant Lectins; Polycystic Kidney Diseases; Proliferating Cell Nuclear Antigen; Regeneration; Renal Dialysis; Soybean Proteins; Uromodulin; Vimentin

The pathogenesis of multicystic dysplastic kidney disease (MCDKD) is unknown. Most morphologic studies of MCDKD kidneys have been performed when the kidneys are resected postnatally, when their architecture has been distorted by massive cyst enlargement. We obtained two MCDKD kidneys at an early stage of development (14 and 19 weeks' gestation) and examined the pattern of nephrogenesis in detail. In both affected kidneys, we identified islands of spatially dislocated metanephric blastema adjacent to zones containing all the normal structural elements of nephrogenesis, including aggregates of induced mesenchyme, S-shaped bodies and maturing glomerull, and proximal and distal tubules. Metanephric blastemal cells displayed characteristic vimentin and smooth muscle actin immunoreactivity and insulin-like growth factor II gene expression, whereas induced elements exhibited appropriate cytokeratin immunoreactivity and Wilms' tumor gene expression. In most other zones, renal cysts were lined with epithelia varying from a flattened squamous to a cuboidal morphology and expression of markers suggested their origin to be from all portions of the nephron including Bowman's space, proximal tubule, and collecting duct. In some cysts, small clusters of epithelial cells were identified within the cyst lumen. These studies suggest that in the early stages of MCDKD, normal nephrogenesis occurs in what seems to be a normal metanephric blastema; however, an intrinsic abnormality in the branching morphogenesis of the ureteric duct might be responsible for the development of the histopathologic changes described.

Topics: Actins; DNA-Binding Proteins; Fetal Diseases; Fetus; Genes, Wilms Tumor; Gestational Age; Humans; Immunohistochemistry; Insulin-Like Growth Factor II; Keratins; Kidney; Kidney Glomerulus; Kidney Tubules; Morphogenesis; Polycystic Kidney Diseases; RNA, Messenger; Transcription Factors; Vimentin; WT1 Proteins

Polycystic kidney disease (PKD), in which epithelial cysts arise from or instead of normal renal tubules, is one of the most common genetic diseases. It has both autosomal dominant and autosomal recessive inheritance in humans and in experimental animals. Epithelial cells lining the cysts have an increased rate of proliferation, abnormal polarity of Na-K-adenosinetriphosphatase, which is localized to apical and sometimes lateral membrane domains, and an abnormal extracellular matrix. One hypothesis that explains the simultaneous acquisition of these characteristics as the result of several different genetic mutations is that cell-matrix interactions, which are known to regulate cell proliferation and cell polarity, are altered in PKD. I have created immortalized renal epithelial cell lines from C57Bl/6Jcpk mice with PKD, an autosomal recessive trait in these animals, and from their phenotypically normal littermates. Using these cell lines, I show that polycystic cells have increased adhesion to collagens and laminin mediated by an integrin. These results demonstrate that cell-matrix interactions are defective in PKD and suggest that these interactions may be involved in the abnormalities of cell polarity and cell proliferation seen in these disorders.

Topics: Animals; Antigens, Viral, Tumor; Blotting, Western; Cadherins; Cell Adhesion; Cell Division; Cell Line; Cell Line, Transformed; Cell Polarity; Collagen; Epithelium; Gene Expression; Immunohistochemistry; Integrins; Keratins; Laminin; Lectins; Mice; Mice, Inbred C57BL; Polycystic Kidney Diseases

Acanthosis nigricans (AN) comprises a broad spectrum of etiologic subtypes. The underlying pathomechanisms have not yet been completely clarified. We present a patient affected with a syndromelike AN subtype including disturbed epidermopoiesis as evidenced by immunohistologic findings and in situ hybridization.. A 54-year-old white man contracted AN during childhood. There were connate malformations consisting of webbed toes II/III on the right side and a supernumerary left mammilla. As an adult he developed psoriasis vulgaris, obesity, and latent diabetes mellitus, polycystic kidney and liver disease. With regard to keratin 6 mRNA, and the protein expression of keratin 6/16, KI-67, and proliferating cell nuclear antigen, the AN lesion showed moderate hyperproliferation. A much higher degree of hyperproliferation was evident in psoriatic areas of the patient's skin. In contrast to psoriatic tissue, basal keratinocytes of the AN showed an unusually high expression of keratin 18 and 19 protein.. The observation thus deals with a unique, syndromelike constellation of AN characterized by a particular epidermal pattern of moderate hyperproliferation. A further dysregulation of protein expression in the epidermis is indicated by the demonstration of the rare keratins 18 and 19 in basal keratinocytes of the AN lesion.

Topics: Abnormalities, Multiple; Acanthosis Nigricans; Humans; Keratins; Male; Middle Aged; Nipples; Polycystic Kidney Diseases; RNA, Messenger; Syndactyly; Syndrome

Progressive renal enlargement is a prominent feature in autosomal dominant polycystic kidney disease (ADPKD), suggesting that the disease is due to hyperplasia and/or preneoplastic transformation of renal epithelial cells. In this study in vitro methods were developed to grow and propagate large numbers of cyst-derived epithelial cells from ADPKD kidneys and cortical epithelial cells from normal human kidneys (NK). In order to study their biologic features during early cell passages, cells were grown on Vitrogen (bovine dermal collagen)-FCS (fetal calf serum) coated dishes and fed a basic medium (DME:F12) supplemented with 10% FCS or a defined medium (Sens) containing insulin, transferrin, selenium, hydrocortisone, tri-iodothyronine and epidermal growth factor (EGF). Both ADPKD and NK cells grew as monolayers, were positive for keratin by immunohistochemistry and flow cytometry and had ultrastructural features of renal epithelial cells. Confluent NK and ADPKD monolayers formed domes. In contrast to NK cells, the growth and propagation of ADPKD cells were not supported by defined medium alone but required serum supplementation and ADPKD cells did not respond to growth factors (insulin, transferrin, EGF) that promoted the growth of NK cells. In serum supplemented media, the growth rate, cell doubling time and end cell number of ADPKD and NK cells were the same. Moreover, ADPKD cells did not exhibit any in vitro features of transformed cells: they were not immortal, they were sensitive to contact inhibition, they were anchorage dependent and they were not tumorigenic in nude mice. These findings do not support an increased rate of cell growth or cell transformation as causative factors in ADPKD.

Topics: Adult; Cell Division; Cells, Cultured; Culture Media; Culture Techniques; Flow Cytometry; Growth Substances; Humans; Immunoenzyme Techniques; Keratins; Kidney; Microscopy, Electron; Middle Aged; Polycystic Kidney Diseases; Reference Values

Human polycystic kidney disease (PKD) epithelia were successfully grown in culture and expressed abnormal characteristics. Cysts lining epithelia of superficial and deep cysts were microdissected and compared to individual normal human proximal straight tubules (PST) and cortical collecting tubules (CCT) grown in defined media. PKD cyst epithelia differed from normal renal tubular epithelia in growth patterns and structural and functional properties. PKD epithelia grew more rapidly and showed cyst-like areas in otherwise confluent monolayers. Polygonal and elongate cells contained an epithelial-specific cytokeratin antigen and had polarized morphology. An extremely abnormal basement membrane morphology was seen and consisted of some banded collagen and numerous unique blebs or spheroids. These blebs were apparently extruded from intracellular vacuoles and stained with ruthenium red, suggesting a proteoglycan component. Cytochemistry of marker enzymes demonstrated the presence of NaK-ATPase and alkaline phosphatase, but a lack of gamma-glutamyl transpeptidase. The response of adenylate cyclase activity to vasopressin, parathyroid hormone, and forskolin was significantly diminished in PKD cells as compared to PST and CCT. These studies suggest a defect in cell growth and basement membrane synthesis in human PKD. Cultured PKD epithelia provide a new tool for the study of the pathogenesis of this disease.

Topics: Adolescent; Adult; Cells, Cultured; Cytological Techniques; Epithelium; Histocytochemistry; Humans; Keratins; Kidney; Kidney Failure, Chronic; Microscopy, Electron; Polycystic Kidney Diseases

Topics: Female; Humans; Infant; Keratins; Kidney; Kidney Neoplasms; Polycystic Kidney Diseases; Wilms Tumor