Efemp1 modulates elastic fiber formation and mechanics of the extrahepatic bile duct

AbstractEGF-Containing Fibulin Extracellular Matrix Protein 1 (EFEMP1, also called fibulin 3) is an extracellular matrix protein linked in a genome-wide association study to biliary atresia, a fibro-inflammatory disease of the neonatal extrahepatic bile duct. EFEMP1 is expressed in most tissues and Efemp1 null mice have decreased elastic fibers in visceral fascia; however, in contrast to other short fibulins (fibulins 4 and 5), EFEMP1 does not have a role in the development of large elastic fibers, and its overall function remains unclear. We demonstrated that EFEMP1 is expressed in the submucosa of both neonatal and adult mouse and human extrahepatic bile ducts and that, in adult Efemp1+/- mice, elastin organization into fibers is decreased. We used pressure myography, a technique developed to study the mechanics of the vasculature, to show that Efemp1+/- extrahepatic bile ducts are more compliant to luminal pressure, leading to increased circumferential stretch. We conclude that EFEMP1 has an important role in the formation of elastic fibers and mechanical properties of the extrahepatic bile duct. These data suggest that altered expression of EFEMP1 in the extrahepatic bile duct leads to an abnormal response to mechanical stress such as obstruction, potentially explaining the role of EFEMP1 in biliary atresia.

Download Full-text

Coordinated development of the mouse extrahepatic bile duct: implications for neonatal susceptibility to biliary injury

10.1101/576256 ◽

2019 ◽

Cited By ~ 1

Author(s):

Gauri Khandekar ◽

Jessica Llewellyn ◽

Alyssa Kriegermeier ◽

Orith Waisbourd-Zinman ◽

Nicolette Johnson ◽

...

Keyword(s):

Bile Acid ◽

Bile Duct ◽

Biliary Atresia ◽

Bile Ducts ◽

Extrahepatic Bile Duct ◽

Collagen I ◽

Cell Junctions ◽

Reporter Mouse ◽

Extrahepatic Bile Ducts

AbstractBackground & AimsThe extrahepatic bile duct is the primary tissue initially affected by the cholangiopathy biliary atresia. Biliary atresia affects neonates exclusively and current animal models suggest that the developing bile duct is uniquely susceptible to damage. In this study, we aimed to define the anatomical and functional differences between the neonatal and adult mouse extrahepatic bile ducts.MethodsWe studied mouse passaged cholangiocytes, mouse BALB/c neonatal and adult primary cholangiocytes and isolated extrahepatic bile ducts, and a collagen reporter mouse. Methods included transmission electron microscopy, lectin staining, immunostaining, rhodamine uptake assays, bile acid toxicity assays, and in vitro modeling of the matrix.ResultsThe cholangiocyte monolayer of the neonatal extrahepatic bile duct was immature, lacking the uniform apical glycocalyx and mature cell-cell junctions typical of adult cholangiocytes. Functional studies showed that the glycocalyx protected against bile acid injury and that neonatal cholangiocyte monolayers were more permeable than adult monolayers. In adult ducts, the submucosal space was filled with collagen I, elastin, hyaluronic acid, and proteoglycans. In contrast, the neonatal submucosa had little collagen I and elastin, although both increased rapidly after birth. In vitro modeling suggested that the composition of the neonatal submucosa relative to the adult submucosa led to increased diffusion of bile. A Col-GFP reporter mouse showed that cells in the neonatal but not adult submucosa were actively producing collagen.ConclusionWe identified four key differences between the neonatal and adult extrahepatic bile duct. We showed that these features may have functional implications, suggesting the neonatal extrahepatic bile ducts are particularly susceptible to injury and fibrosis.Lay SummaryBiliary atresia is a disease that affects newborns and is characterized by extrahepatic bile duct injury and obstruction with resulting liver injury. We identify four key differences between the epithelial and submucosal layers of the neonatal and adult extrahepatic bile duct and show that these may render the neonatal duct particularly susceptible to injury.HighlightsThe apical glycocalyx is thin and patchy in neonatal compared to adult cholangiocytesNeonatal cholangiocytes have immature cell-cell junctions and increased permeabilityThe neonatal submucosal space has minimal collagen I or elastinThe neonatal submucosal space contains many actively collagen-secreting cellsGraphical abstract

Download Full-text

Proximal Extrahepatic Bile Ducts: Comprehensive Review

Journal of oncology: diagnostic radiology and radiotherapy ◽

10.37174/2587-7593-2021-4-1-74-93 ◽

2021 ◽

Vol 4 (1) ◽

pp. 74-93

Author(s):

M. A. Shorikov ◽

O. N. Sergeeva ◽

M. G. Lapteva ◽

N. A. Peregudov ◽

B. I. Dolgushin

Keyword(s):

Bile Duct ◽

Bile Flow ◽

Bile Ducts ◽

Extrahepatic Bile Duct ◽

Hepatic Duct ◽

Biliary Tree ◽

Comprehensive Review ◽

Extrahepatic Bile Ducts ◽

And Function ◽

Variant Anatomy

Proximal extrahepatic bile ducts are the biliary tree segment within formal boundaries from cystic ductcommon hepatic duct junction to sectoral hepatic ducts. Despite being a focus of attention of diagnostic and interventional radiologists, endoscopists, hepatobiliary surgeons and transplantologists they weren’t comprehensively described in available papers. The majority of the authors regard bile duct confluence as a group of merging primitively arranged tubes providing bile flow. The information on the proximal extrahepatic bile duct embryonal development, variant anatomy, innervation, arterial, venous and lymphatic supply is too general and not detailed. The present review brought together and systemized exiting to the date data on anatomy and function of this biliary tract portion. Unique, different from the majority of hollow organs organization of the proximal extrahepatic bile duct adapts them to the flow of the bile, i.e. viscous aggressive due to pH about 8.0 and detergents fluid, under higher wall pressure than in other parts of biliary tree.

Download Full-text

Expression of the extracellular matrix protein periostin in liver tumours and bile duct carcinomas

Histopathology ◽

10.1111/j.1365-2559.2010.03527.x ◽

2010 ◽

Vol 56 (5) ◽

pp. 600-606 ◽

Cited By ~ 39

Author(s):

Marc-Oliver Riener ◽

Florian R Fritzsche ◽

Christopher Soll ◽

Bernhard C Pestalozzi ◽

Nicole Probst-Hensch ◽

...

Keyword(s):

Extracellular Matrix ◽

Bile Duct ◽

Matrix Protein ◽

Extracellular Matrix Protein ◽

Liver Tumours

Download Full-text

Characterization of Cardiac Function and Arterial Mechanics During Early Postnatal Development in Fibulin-5 Null Mice

Volume 1B: Extremity; Fluid Mechanics; Gait; Growth, Remodeling, and Repair; Heart Valves; Injury Biomechanics; Mechanotransduction and Sub-Cellular Biophysics; MultiScale Biotransport; Muscle, Tendon and Ligament; Musculoskeletal Devices; Multiscale Mechanics; Thermal Medicine; Ocular Biomechanics; Pediatric Hemodynamics; Pericellular Phenomena; Tissue Mechanics; Biotransport Design and Devices; Spine; Stent Device Hemodynamics; Vascular Solid Mechanics; Student Paper and Design Competitions ◽

10.1115/sbc2013-14282 ◽

2013 ◽

Author(s):

Victoria Le ◽

Hiromi Yanagisawa ◽

Jessica Wagenseil

Keyword(s):

Matrix Protein ◽

Elastic Fiber ◽

Connective Tissue Disorder ◽

Fiber Formation ◽

Extracellular Matrix Protein ◽

Elastic Fibers ◽

Arterial Mechanics ◽

Early Postnatal Development ◽

Complex Process

Fibulin-5 is an extracellular matrix protein that interacts with other proteins during a complex process that results in elastic fiber formation from the elastin precursor, tropoelastin [1]. Elastic fibers are an important component of tissues requiring elasticity, including large arteries, lungs and skin. In mice lacking fibulin-5 ( Fbln5−/−), these tissues contain disorganized elastic fibers and exhibit decreased elasticity [2]. The phenotype of Fbln5−/− mice is similar to that of humans with cutis laxa, a connective tissue disorder characterized by loose skin and narrow arteries with reduced compliance.

Download Full-text

Cholangioscopy in diagnosis of diseases of extrahepatic bile ducts

Medical alphabet ◽

10.33667/2078-5631-2019-1-6(381)-30-36 ◽

2019 ◽

Vol 1 (6) ◽

pp. 30-36

Author(s):

A. G. Shuleshov ◽

N. V. Fomicheva ◽

D. N. Ulyanov ◽

A. S. Balalykin ◽

D. V. Danilov ◽

...

Keyword(s):

Differential Diagnosis ◽

Bile Duct ◽

Bile Ducts ◽

Extrahepatic Bile Duct ◽

Diagnostic Purpose ◽

Vascular Pattern ◽

Direct Visualization ◽

Bile Duct Diseases ◽

Extrahepatic Bile Ducts ◽

Diagnosis Of Diseases

An analysis of the diagnosis of extrahepatic bile duct diseases in 115 patients is presented. With the diagnostic purpose they performed ERCP, EPT, cholangioscopy. The method of direct visualization of the mucous membrane of the bile ducts allows you to identify endoscopic signs of strictures. Differential diagnosis of benign and malignant strictures of the bile ducts using cholangioscopy is difficult. Nevertheless, we were able to identify some typical signs for malignant strictures, including ulceration and mucosal infiltration, vascular pattern irregularity, stricture asymmetry.

Download Full-text

Developmental Regulation of FKBP65

Molecular Biology of the Cell ◽

10.1091/mbc.11.11.3925 ◽

2000 ◽

Vol 11 (11) ◽

pp. 3925-3935 ◽

Cited By ~ 47

Author(s):

Charles E. Patterson ◽

Theresa Schaub ◽

Elaine J. Coleman ◽

Elaine C. Davis

Keyword(s):

Extracellular Matrix ◽

Matrix Protein ◽

Subcellular Fractionation ◽

Immunohistochemical Localization ◽

Extracellular Matrix Protein ◽

Elastic Fibers ◽

Airway Smooth Muscle Cells ◽

Specific Expression ◽

Adult Tissues ◽

Mouse Tissues

FKBP65 (65-kDa FK506-binding protein) is a member of the highly conserved family of intracellular receptors called immunophilins. All have the property of peptidyl-prolyl cis-trans isomerization, and most have been implicated in folding and trafficking events. In an earlier study, we identified that FKBP65 associates with the extracellular matrix protein tropoelastin during its transport through the cell. In the present study, we have carried out a detailed investigation of the subcellular localization of FKBP65 and its relationship to tropoelastin. Using subcellular fractionation, Triton X-114 phase separation, protease protection assays, and immunofluorescence microscopy (IF), we have identified that FKBP65 is contained within the lumen of the endoplasmic reticulum (ER). Subsequent IF studies colocalized FKBP65 with tropoelastin and showed that the two proteins dissociate before reaching the Golgi apparatus. Immunohistochemical localization of FKBP65 in developing lung showed strong staining of vascular and airway smooth muscle cells. Similar areas stained positive for the presence of elastic fibers in the extracellular matrix. The expression of FKBP65 was investigated during development as tropoelastin is not expressed in adult tissues. Tissue-specific expression of FKBP65 was observed in 12-d old mouse tissues; however, the pattern of expression of FKBP65 was not restricted to those tissues expressing tropoelastin. This suggests that additional ligands for FKBP65 likely exist within the ER. Remarkably, in the adult tissues examined, FKBP65 expression was absent or barely detectable. Taken together, these results support an ER-localized FKBP65-tropoelastin interaction that occurs specifically during growth and development of tissues.

Download Full-text

Role of LTBP4 in alveolarization, angiogenesis, and fibrosis in lungs

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00031.2017 ◽

2017 ◽

Vol 313 (4) ◽

pp. L687-L698 ◽

Cited By ~ 8

Author(s):

Insa Bultmann-Mellin ◽

Katharina Dinger ◽

Carolin Debuschewitz ◽

Katharina M. A. Loewe ◽

Yvonne Melcher ◽

...

Keyword(s):

Extracellular Matrix ◽

Matrix Protein ◽

Transforming Growth Factor ◽

Elastic Fiber ◽

Transforming Growth Factor Β ◽

Matrix Proteins ◽

Extracellular Matrix Protein ◽

Elastic Fibers ◽

Fiber Network

Deficiency of the extracellular matrix protein latent transforming growth factor-β (TGF-β)-binding protein-4 (LTBP4) results in lack of intact elastic fibers, which leads to disturbed pulmonary development and lack of normal alveolarization in humans and mice. Formation of alveoli and alveolar septation in pulmonary development requires the concerted interaction of extracellular matrix proteins, growth factors such as TGF-β, fibroblasts, and myofibroblasts to promote elastogenesis as well as vascular formation in the alveolar septae. To investigate the role of LTBP4 in this context, lungs of LTBP4-deficient ( Ltbp4−/−) mice were analyzed in close detail. We elucidate the role of LTBP4 in pulmonary alveolarization and show that three different, interacting mechanisms might contribute to alveolar septation defects in Ltbp4−/− lungs: 1) absence of an intact elastic fiber network, 2) reduced angiogenesis, and 3) upregulation of TGF-β activity resulting in profibrotic processes in the lung.

Download Full-text

Perfusion Decellularization of Extrahepatic Bile Duct Allows Tissue-Engineered Scaffold Generation by Preserving Matrix Architecture and Cytocompatibility

Materials ◽

10.3390/ma14113099 ◽

2021 ◽

Vol 14 (11) ◽

pp. 3099

Author(s):

Yolik Ramírez-Marín ◽

David Eduardo Abad-Contreras ◽

Martha Ustarroz-Cano ◽

Norma S. Pérez-Gallardo ◽

Lorena Villafuerte-García ◽

...

Keyword(s):

Extracellular Matrix ◽

Bile Duct ◽

Bile Ducts ◽

Extrahepatic Bile Duct ◽

Hierarchical Structures ◽

Human Leukocyte ◽

Medical Problem ◽

Biochemical Properties ◽

Biological Scaffolds ◽

Ultrastructure Preservation

Reconstruction of bile ducts damaged remains a vexing medical problem. Surgeons have few options when it comes to a long segment reconstruction of the bile duct. Biological scaffolds of decellularized biliary origin may offer an approach to support the replace of bile ducts. Our objective was to obtain an extracellular matrix scaffold derived from porcine extrahepatic bile ducts (dECM-BD) and to analyze its biological and biochemical properties. The efficiency of the tailored perfusion decellularization process was assessed through histology stainings. Results from 4’-6-diamidino-2-phenylindole (DAPI), Hematoxylin and Eosin (H&E) stainings, and deoxyribonucleic acid (DNA) quantification showed proper extracellular matrix (ECM) decellularization with an effectiveness of 98%. Immunohistochemistry results indicate an effective decrease in immunogenic marker as human leukocyte antigens (HLA-A) and Cytokeratin 7 (CK7) proteins. The ECM of the bile duct was preserved according to Masson and Herovici stainings. Data derived from scanning electron microscopy (SEM) and thermogravimetric analysis (TGA) showed the preservation of the dECM-BD hierarchical structures. Cytotoxicity of dECM-BD was null, with cells able to infiltrate the scaffold. In this work, we standardized a decellularization method that allows one to obtain a natural bile duct scaffold with hierarchical ultrastructure preservation and adequate cytocompatibility.

Download Full-text

Fibulin-2 Is Dispensable for Mouse Development and Elastic Fiber Formation

Molecular and Cellular Biology ◽

10.1128/mcb.01876-07 ◽

2007 ◽

Vol 28 (3) ◽

pp. 1061-1067 ◽

Cited By ~ 44

Author(s):

Francois-Xavier Sicot ◽

Takeshi Tsuda ◽

Dessislava Markova ◽

John F. Klement ◽

Machiko Arita ◽

...

Keyword(s):

Matrix Protein ◽

Elastic Fiber ◽

Embryonic Stem ◽

Fiber Formation ◽

Extracellular Matrix Protein ◽

Elastic Fibers ◽

Mouse Development ◽

Fibrillin 1

ABSTRACT Fibulin-2 is an extracellular matrix protein belonging to the five-member fibulin family, of which two members have been shown to play essential roles in elastic fiber formation during development. Fibulin-2 interacts with two major constituents of elastic fibers, tropoelastin and fibrillin-1, in vitro and localizes to elastic fibers in many tissues in vivo. The protein is prominently expressed during morphogenesis of the heart and aortic arch vessels and at early stages of cartilage development. To examine its role in vivo, we generated mice that do not express the fibulin-2 gene (Fbln2) through homologous recombination of embryonic stem cells. Unexpectedly, the fibulin-2-null mice were viable and fertile and did not display gross and anatomical abnormalities. Histological and ultrastructural analyses revealed that elastic fibers assembled normally in the absence of fibulin-2. No compensatory up-regulation of mRNAs for other fibulin members was detected in the aorta and skin tissue. However, in the fibulin-2 null aortae, fibulin-1 immunostaining was increased in the inner elastic lamina, where fibulin-2 preferentially localizes. The results demonstrate that fibulin-2 is not required for mouse development and elastic fiber formation and suggest possible functional redundancy between fibulin-1 and fibulin-2.

Download Full-text