Three-dimensional structural analysis reveals a Cdk5-mediated kinase cascade regulating hepatic biliary network branching in zebrafish

Dimri, Manjari; Bilogan, Cassandra K.; Pierce, Lain X.; Naegele, Gregory; Vasanji, Amit; Gibson, Isabel; McClendon, Allyson; Tae, Kevin; Sakaguchi, Takuya

doi:10.1242/jcs.208702

Cited by 3 publications

(16 citation statements)

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“…Sections were mounted with ProLong Gold Antifade Mountant with 4′,6‐diamidino‐2‐phenylindole (DAPI; Life Technologies) and imaged using a Leica SP5 DMI at 63×. Biliary network reconstruction was performed as described . Mouse liver sections (Lrat‐CRE/ZsGreen Flox/Flox ) were fixed in 4% PFA in PBS overnight and incubated for 24 hours each in 10%, 20%, and then 30% sucrose in PBS before embedding in OCT, and sectioned in 10 µM.…”

Section: Methodsmentioning

confidence: 99%

“…Biliary network reconstruction was performed as described. (14) Mouse liver sections (Lrat-CRE/ZsGreen Flox/Flox ) (15) were fixed in 4% PFA in PBS overnight and incubated for 24 hours each in 10%, 20%, and then 30% sucrose in PBS before embedding in OCT, and sectioned in 10 µM. Tissue sections were rehydrated in PBS and incubated with anti-MPI antibody (Sigma SAB2700835, 1:100; Sigma-Aldrich, St. Louis, MO), anti-CD31 antibody (550274, 1:200; BD Pharmingen, San Jose, CA), and anti-glial fibrillary acidic protein (GFAP; ab4674, 1:200; Abcam, Cambridge, MA) diluted in PBS overnight at 4˚C.…”

Section: Immunofluorescencementioning

confidence: 99%

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Mannose Phosphate Isomerase and Mannose Regulate Hepatic Stellate Cell Activation and Fibrosis in Zebrafish and Humans

et al. 2019

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The growing burden of liver fibrosis and lack of effective antifibrotic therapies highlight the need for identification of pathways and complementary model systems of hepatic fibrosis. A rare, monogenic disorder in which children with mutations in mannose phosphate isomerase (MPI) develop liver fibrosis led us to explore the function of MPI and mannose metabolism in liver development and adult liver diseases. Herein, analyses of transcriptomic data from three human liver cohorts demonstrate that MPI gene expression is down‐regulated proportionate to fibrosis in chronic liver diseases, including nonalcoholic fatty liver disease and hepatitis B virus. Depletion of MPI in zebrafish liver in vivo and in human hepatic stellate cell (HSC) lines in culture activates fibrotic responses, indicating that loss of MPI promotes HSC activation. We further demonstrate that mannose supplementation can attenuate HSC activation, leading to reduced fibrogenic activation in zebrafish, culture‐activated HSCs, and in ethanol‐activated HSCs. Conclusion: These data indicate the prospect that modulation of mannose metabolism pathways could reduce HSC activation and improve hepatic fibrosis.

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Section: Methodsmentioning

confidence: 99%

Section: Immunofluorescencementioning

confidence: 99%

Mannose Phosphate Isomerase and Mannose Regulate Hepatic Stellate Cell Activation and Fibrosis in Zebrafish and Humans

et al. 2019

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“…D, arrows, percentage of C5 – bile ductules displaying C5 fluorescence at both ends), suggesting a defect in the formation of open ductules. Given the essential role of filamentous actin (F‐actin) assembly in lumen and bile conduit formation, we examined actin filaments in XAV939‐treated livers. Although no difference in BEC morphology (i.e., Tp1 :GFP expression) was observed between XAV939‐treated and control livers, the F‐actin filaments appeared disconnected in XAV939‐treated livers (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Intrahepatic biliary morphogenesis continues between 4 and 5 dpf: bile ductules further elongate and become thinner . However, suppressing Wnt/β‐catenin signaling during this period did not impair intrahepatic biliary morphogenesis or reduce Notch activity (http://onlinelibrary.wiley.com/doi/10.1002/hep.29752/suppinfo).…”

Section: Discussionmentioning

confidence: 99%

Wnt/β‐catenin signaling controls intrahepatic biliary network formation in zebrafish by regulating notch activity

Khaliq

Evason

et al. 2018

Hepatology

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“…This transgenic line allowed us to visualize the entire intrahepatic biliary network in zebrafish larvae, greatly facilitating investigation of the development of the biliary system. Utilizing this transgenic line, we have previously developed computational algorithms to convert complex three-dimensional network structures into arrays of numeric sub-parameters to quantify the subtle differences in intrahepatic biliary network branching patterns (16). Precise computational quantification of the intrahepatic biliary network enabled us to unbiasedly quantify mutant phenotypes and break down complicated three-dimensional branching patterns into multi-dimensional arrays, which we can feed into unsupervised classification algorithms.…”

Section: Introductionmentioning

confidence: 99%

Forward genetics combined with unsupervised classifications identified zebrafish mutants affecting biliary system formation

et al. 2021

Preprint

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Impaired formation of the biliary network can lead to congenital cholestatic liver diseases; however, the genes responsible for proper biliary system formation and maintenance have not been fully identified. Combining computational network structure analysis algorithms with a zebrafish forward genetic screen, we identified 24 new zebrafish mutants that display impaired intrahepatic biliary network formation. Complementation tests suggested that these 24 mutants affect 24 different genes. We applied unsupervised clustering algorithms to classify the recovered mutants into three classes unbiasedly. Further computational analyses revealed that each of the recovered mutations in these three classes shows a unique effect on node subtype composition and connection property distribution of the intrahepatic biliary network. Besides, we found that most recovered mutations are viable. In those mutant fish, biliary network phenotypes persist into adulthood, which themselves are good animal models to study chronic cholestatic liver diseases. Altogether, this study provides unique genetic and computational toolsets that advance our understanding of the molecular pathways leading to biliary system malformation and cholestatic liver diseases.

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Three-dimensional structural analysis reveals a Cdk5-mediated kinase cascade regulating hepatic biliary network branching in zebrafish

Cited by 3 publications

References 1 publication

Mannose Phosphate Isomerase and Mannose Regulate Hepatic Stellate Cell Activation and Fibrosis in Zebrafish and Humans

Mannose Phosphate Isomerase and Mannose Regulate Hepatic Stellate Cell Activation and Fibrosis in Zebrafish and Humans

Wnt/β‐catenin signaling controls intrahepatic biliary network formation in zebrafish by regulating notch activity

Forward genetics combined with unsupervised classifications identified zebrafish mutants affecting biliary system formation

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