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

Gauri Khandekar, Jessica Llewellyn, Alyssa Kriegermeier, Orith Waisbourd-Zinman, Nicolette Johnson, Yu Du, Roquibat Giwa, Xiao Liu, Tatiana Kisseleva, Pierre A. Russo, Neil D. Theise, Rebecca G. Wells*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Background & Aims: The extrahepatic bile duct is the primary tissue initially affected by biliary atresia. Biliary atresia is a cholangiopathy which exclusively affects neonates. 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. Methods: We studied mouse passaged cholangiocytes, mouse BALB/c neonatal and adult primary cholangiocytes, as well as isolated extrahepatic bile ducts, and a collagen reporter mouse. The methods used included transmission electron microscopy, lectin staining, immunostaining, rhodamine uptake assays, bile acid toxicity assays, and in vitro modeling of the matrix. Results: The 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 of the matrix 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. Conclusion: We identified 4 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 summary: Biliary atresia is a disease that affects newborns and is characterized by extrahepatic bile duct injury and obstruction, resulting in liver injury. We identify 4 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.

Original languageEnglish (US)
Pages (from-to)135-145
Number of pages11
JournalJournal of Hepatology
Volume72
Issue number1
DOIs
StatePublished - Jan 2020

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Extrahepatic Bile Ducts
Biliary Atresia
Wounds and Injuries
Collagen
Glycocalyx
Elastin
Bile Acids and Salts
Extrahepatic Cholestasis
Rhodamines
Intercellular Junctions
Proteoglycans
Hyaluronic Acid
Bile Ducts
Transmission Electron Microscopy
Lectins
Bile
Fibrosis
Animal Models
Parturition
Staining and Labeling

Keywords

  • Biliary atresia
  • Fibrogenesis
  • Glycocalyx
  • Lectin
  • Submucosa

ASJC Scopus subject areas

  • Hepatology

Cite this

Khandekar, Gauri ; Llewellyn, Jessica ; Kriegermeier, Alyssa ; Waisbourd-Zinman, Orith ; Johnson, Nicolette ; Du, Yu ; Giwa, Roquibat ; Liu, Xiao ; Kisseleva, Tatiana ; Russo, Pierre A. ; Theise, Neil D. ; Wells, Rebecca G. / Coordinated development of the mouse extrahepatic bile duct : Implications for neonatal susceptibility to biliary injury. In: Journal of Hepatology. 2020 ; Vol. 72, No. 1. pp. 135-145.
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abstract = "Background & Aims: The extrahepatic bile duct is the primary tissue initially affected by biliary atresia. Biliary atresia is a cholangiopathy which exclusively affects neonates. 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. Methods: We studied mouse passaged cholangiocytes, mouse BALB/c neonatal and adult primary cholangiocytes, as well as isolated extrahepatic bile ducts, and a collagen reporter mouse. The methods used included transmission electron microscopy, lectin staining, immunostaining, rhodamine uptake assays, bile acid toxicity assays, and in vitro modeling of the matrix. Results: The 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 of the matrix 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. Conclusion: We identified 4 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 summary: Biliary atresia is a disease that affects newborns and is characterized by extrahepatic bile duct injury and obstruction, resulting in liver injury. We identify 4 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.",
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author = "Gauri Khandekar and Jessica Llewellyn and Alyssa Kriegermeier and Orith Waisbourd-Zinman and Nicolette Johnson and Yu Du and Roquibat Giwa and Xiao Liu and Tatiana Kisseleva and Russo, {Pierre A.} and Theise, {Neil D.} and Wells, {Rebecca G.}",
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Khandekar, G, Llewellyn, J, Kriegermeier, A, Waisbourd-Zinman, O, Johnson, N, Du, Y, Giwa, R, Liu, X, Kisseleva, T, Russo, PA, Theise, ND & Wells, RG 2020, 'Coordinated development of the mouse extrahepatic bile duct: Implications for neonatal susceptibility to biliary injury', Journal of Hepatology, vol. 72, no. 1, pp. 135-145. https://doi.org/10.1016/j.jhep.2019.08.036

Coordinated development of the mouse extrahepatic bile duct : Implications for neonatal susceptibility to biliary injury. / Khandekar, Gauri; Llewellyn, Jessica; Kriegermeier, Alyssa; Waisbourd-Zinman, Orith; Johnson, Nicolette; Du, Yu; Giwa, Roquibat; Liu, Xiao; Kisseleva, Tatiana; Russo, Pierre A.; Theise, Neil D.; Wells, Rebecca G.

In: Journal of Hepatology, Vol. 72, No. 1, 01.2020, p. 135-145.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Coordinated development of the mouse extrahepatic bile duct

T2 - Implications for neonatal susceptibility to biliary injury

AU - Khandekar, Gauri

AU - Llewellyn, Jessica

AU - Kriegermeier, Alyssa

AU - Waisbourd-Zinman, Orith

AU - Johnson, Nicolette

AU - Du, Yu

AU - Giwa, Roquibat

AU - Liu, Xiao

AU - Kisseleva, Tatiana

AU - Russo, Pierre A.

AU - Theise, Neil D.

AU - Wells, Rebecca G.

PY - 2020/1

Y1 - 2020/1

N2 - Background & Aims: The extrahepatic bile duct is the primary tissue initially affected by biliary atresia. Biliary atresia is a cholangiopathy which exclusively affects neonates. 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. Methods: We studied mouse passaged cholangiocytes, mouse BALB/c neonatal and adult primary cholangiocytes, as well as isolated extrahepatic bile ducts, and a collagen reporter mouse. The methods used included transmission electron microscopy, lectin staining, immunostaining, rhodamine uptake assays, bile acid toxicity assays, and in vitro modeling of the matrix. Results: The 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 of the matrix 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. Conclusion: We identified 4 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 summary: Biliary atresia is a disease that affects newborns and is characterized by extrahepatic bile duct injury and obstruction, resulting in liver injury. We identify 4 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.

AB - Background & Aims: The extrahepatic bile duct is the primary tissue initially affected by biliary atresia. Biliary atresia is a cholangiopathy which exclusively affects neonates. 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. Methods: We studied mouse passaged cholangiocytes, mouse BALB/c neonatal and adult primary cholangiocytes, as well as isolated extrahepatic bile ducts, and a collagen reporter mouse. The methods used included transmission electron microscopy, lectin staining, immunostaining, rhodamine uptake assays, bile acid toxicity assays, and in vitro modeling of the matrix. Results: The 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 of the matrix 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. Conclusion: We identified 4 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 summary: Biliary atresia is a disease that affects newborns and is characterized by extrahepatic bile duct injury and obstruction, resulting in liver injury. We identify 4 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.

KW - Biliary atresia

KW - Fibrogenesis

KW - Glycocalyx

KW - Lectin

KW - Submucosa

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