Actomyosin contractility drives bile regurgitation as an early response during obstructive cholestasis
Open accessPublished version of a preprint
Abstract
Background & aims A wide range of liver diseases manifest as biliary obstruction, or cholestasis. However, the sequence of molecular events triggered as part of the early hepatocellular homeostatic response in obstructive cholestasis is poorly elucidated. Pericanalicular actin is known to accumulate during obstructive cholestasis. Therefore, we hypothesized that the pericanalicular actin cortex undergoes significant remodeling as a regulatory response to obstructive cholestasis.Methods In vivo investigations were performed in a bile duct-ligated mouse model. Actomyosin contractility was assessed using sandwich-cultured rat hepatocytes transfected with various fluorescently labeled proteins and pharmacological inhibitors of actomyosin contractility.Results Actomyosin contractility induces transient deformations along the canalicular membrane, a process we have termed inward blebbing. We show that these membrane intrusions are initiated by local ruptures in the pericanalicular actin cortex; and they typically retract following repair by actin polymerization and actomyosin contraction. However, above a certain osmotic pressure threshold, these inward blebs pinch away from the canalicular membrane into the hepatocyte cytoplasm as large vesicles (2-8μm). Importantly, we show that these vesicles aid in the regurgitation of bile from the bile canaliculi.Conclusion Actomyosin contractility induces the formation of bile-regurgitative vesicles, thus serving as an early homeostatic mechanism against increased biliary pressure during cholestasis.Lay summary Bile canaliculi expand and contract in response to the amount of secreted bile, and resistance from the surrounding actin bundles. Further expansion due to bile duct blockade leads to the formation of inward blebs, which carry away excess bile to prevent bile build up in the canaliculi.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Regulatory hardstops are now established on multiple fronts, rapidly changing the situation.
“However, this is rapidly changing with regulatory hardstops now established on multiple fronts252.”
From Organoids (2022) · cites this paper as [252]
The Results and Discussion passages describe bile-canalicular dynamics and bile regurgitation during obstructive cholestasis, not regulatory measures. Searches for “regulatory hardstops” and regulatory barriers or enforcement found no passage addressing the claim; page numbers were not available in our copy. (LLM, conf=0.78)
Experimental — accuracy not yet measured
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Chapter 4 Preparation of Isolated Rat Liver Cells
Seglen, P. O. (1976) · Methods in cell biology · doi:10.1016/s0091-679x(08)61797-5 · Cited by 5,297 · see citing works
α-Catenin as a tension transducer that induces adherens junction development
Yonemura, S., Wada, Y., Watanabe, T., Nagafuchi, A., & Shibata, M. (2010) · Nature Cell Biology · doi:10.1038/ncb2055 · Cited by 972 · see citing works
Hartley, J., Davenport, M., & Kelly, D. A. (2009) · The Lancet · doi:10.1016/s0140-6736(09)60946-6 · Cited by 880 · see citing works
Blebs lead the way: how to migrate without lamellipodia
Charras, G., & Paluch, E. K. (2008) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm2453 · Cited by 776 · see citing works
Bile-acid-induced cell injury and protection
Perez, M. J., & Briz, O. (2009) · World Journal of Gastroenterology · doi:10.3748/wjg.15.1677 · Cited by 717 · see citing works
Korb, L. C., & Ahearn, J. M. (1997) · The Journal of Immunology · doi:10.4049/jimmunol.158.10.4525 · Cited by 659 · see citing works
Reassembly of contractile actin cortex in cell blebs
Charras, G., Hu, C., Coughlin, M. L., & Mitchison, T. J. (2006) · The Journal of Cell Biology · doi:10.1083/jcb.200602085 · Cited by 620 · see citing works
Role of cortical tension in bleb growth
Tinévez, J.-Y., Schulze, U., Salbreux, G., Roensch, J., Joanny, J., & Paluch, E. K. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0903353106 · Cited by 543 · see citing works
Life and Times of a Cellular Bleb
Charras, G., Coughlin, M. L., Mitchison, T. J., & Mahadevan, L. (2007) · Biophysical Journal · doi:10.1529/biophysj.107.113605 · Cited by 456 · see citing works
Generation of compartmentalized pressure by a nuclear piston governs cell motility in a 3D matrix
Petrie, R. J., Koo, H., & Yamada, K. M. (2014) · Science · doi:10.1126/science.1256965 · Cited by 367 · see citing works
The role and regulation of blebs in cell migration
Paluch, E. K., & Raz, E. (2013) · Current Opinion in Cell Biology · doi:10.1016/j.ceb.2013.05.005 · Cited by 365 · see citing works
Lifeact mice for studying F-actin dynamics
Riedl, J., Flynn, K. C., Raducanu, A., Gärtner, F. C., Beck, G., Bösl, M. R., Bradke, F., Maßberg, S., Aszódi, A., Sixt, M., et al. (2010) · Nature Methods · doi:10.1038/nmeth0310-168 · Cited by 333 · see citing works
Charras, G. (2008) · Journal of Microscopy · doi:10.1111/j.1365-2818.2008.02059.x · Cited by 322 · see citing works
Fickert, P., Zollner, G., Fuchsbichler, A., Stumptner, C., Weiglein, A. H., Lammert, F., Marschall, H., Tsybrovskyy, O., Zatloukal, K., Denk, H., et al. (2002) · Gastroenterology · doi:10.1053/gast.2002.35948 · Cited by 312 · see citing works
New molecular insights into the mechanisms of cholestasis.
Wagner, M., Zollner, G., & Trauner, M. (2009) · Journal of Hepatology · doi:10.1016/j.jhep.2009.05.012 · Cited by 270 · see citing works
Cipolla, M. J., Gokina, N. I., & Osol, G. (2002) · The FASEB Journal · doi:10.1096/cj.01-0104hyp · Cited by 237 · see citing works
Ritsma, L., Steller, E. J. A., Beerling, E., Loomans, C. J. M., Zomer, A., Gerlach, C., Vrisekoop, N., Seinstra, D., van Gurp, L., Schäfer, R., et al. (2012) · Science Translational Medicine · doi:10.1126/scitranslmed.3004394 · Cited by 213 · see citing works
Blood flow drives lumen formation by inverse membrane blebbing during angiogenesis in vivo
Gebala, V., Collins, R., Geudens, I., Phng, L., & Gerhardt, H. (2016) · Nature Cell Biology · doi:10.1038/ncb3320 · Cited by 195 · see citing works
Chen, F. Y., Ananthanarayanan, M., Emre, S., Neimark, E., Bull, L. N., Knisely, A. S., Strautnieks, S. S., Thompson, R. J., Magid, M. S., Gordon, R. D., et al. (2004) · Gastroenterology · doi:10.1053/j.gastro.2003.12.013 · Cited by 191 · see citing works
Li, M. K., & Crawford, J. (2004) · Seminars in liver disease (Print) · doi:10.1055/s-2004-823099 · Cited by 173 · see citing works
Crawford, A. R., Smith, A. J., Hatch, V., Elferink, R. P. J. O., Borst, P., & Crawford, J. M. (1997) · Journal of Clinical Investigation · doi:10.1172/jci119799 · Cited by 155 · see citing works
Verma, S., Han, S. P., Michael, M., Gomez, G. A., Yang, Z., Teasdale, R. D., Ratheesh, A., Kovács, É., Ali, R. G., & Yap, A. S. (2012) · Molecular Biology of the Cell · doi:10.1091/mbc.e12-08-0574 · Cited by 144 · see citing works
Functional Anatomy of Normal Bile Ducts
Strazzabosco, M., & Fabris, L. (2008) · The Anatomical Record · doi:10.1002/ar.20664 · Cited by 140 · see citing works
Motility of bile canaliculi in the living animal: implications for bile flow.
Watanabe, N., Tsukada, N., Smith, C. S., & Phillips, M. J. (1991) · The Journal of Cell Biology · doi:10.1083/jcb.113.5.1069 · Cited by 135 · see citing works
Physiological and molecular biochemical mechanisms of bile formation
Reshetnyak, V. I. (2013) · World Journal of Gastroenterology · doi:10.3748/wjg.v19.i42.7341 · Cited by 135 · see citing works
Ultrastructure of the spinal cord of the lamprey
Bertolini, B. (1964) · Journal of Ultrastructure Research · doi:10.1016/s0022-5320(64)80089-7 · Cited by 129 · see citing works
Animal models of portal hypertension
Abraldeṣ, J. G. (2006) · World Journal of Gastroenterology · doi:10.3748/wjg.v12.i41.6577 · Cited by 125 · see citing works
Biliary Atresia: 50 Years after the First Kasai
Wildhaber, B. E. (2012) · ISRN Surgery · doi:10.5402/2012/132089 · Cited by 119 · see citing works
Cholestasis‐induced adaptive remodeling of interlobular bile ducts
Vartak, N., Damle‐Vartak, A., Richter, B., Dirsch, O., Dahmen, U., Hammad, S., & Hengstler, J. G. (2015) · Hepatology · doi:10.1002/hep.28373 · Cited by 115 · see citing works
Localization of P-gp (Abcb1) and Mrp2 (Abcc2) in Freshly Isolated Rat Hepatocytes
Bow, D. A. J., Perry, J. L., Miller, D. S., Pritchard, J. B., & Brouwer, K. L. R. (2007) · Drug Metabolism and Disposition · doi:10.1124/dmd.107.018200 · Cited by 109 · see citing works
Bleb formation and F-actin distribution during mitosis and tumor necrosis factor-induced apoptosis
Laster, S. M., & Mackenzie, J. (1996) · Microscopy Research and Technique · doi:10.1002/(sici)1097-0029(19960615)34:3 · Cited by 99 · see citing works
Fransen, J. H., Hilbrands, L. B., Ruben, J. M., Stoffels, M., Adema, G. J., van der Vlag, J., & Berden, J. H. M. (2009) · Arthritis & Rheumatism · doi:10.1002/art.24719 · Cited by 93 · see citing works
Chen, H., Liu, Y., Chen, H., Wu, S., Ni, Y., Ho, M., Lai, H., Hsu, W., Hsu, H., Tseng, H.-C., et al. (2008) · Pediatric Research · doi:10.1203/pdr.0b013e318170a6b5 · Cited by 91 · see citing works
A role for microtubules in sorting endocytic vesicles in rat hepatocytes.
Goltz, J. S., Wolkoff, A. W., Novikoff, P. M., et al. (1992) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.89.15.7026 · Cited by 87 · see citing works
Synaptic vesicles really do kiss and run
Wightman, R. M., & Haynes, C. L. (2004) · Nature Neuroscience · doi:10.1038/nn0404-321 · Cited by 81 · see citing works
Li, Q., Zhang, Y., Pluchon, P., Robens, J., Herr, K. J., Mercade, M., Thiery, J. P., Yu, H., & Viasnoff, V. (2016) · Nature Cell Biology · doi:10.1038/ncb3310 · Cited by 74 · see citing works
Self-citationThe unique polarity phenotype of hepatocytes
Müsch, A. (2014) · Experimental Cell Research · doi:10.1016/j.yexcr.2014.06.006 · Cited by 64 · see citing works
Lipoprotein–X in Patients With Cirrhosis: Its Relationship to Cholestasis and Hypercholesterolemia
Sörös, P., Böttcher, J., Maschek, H., et al. (1998) · Hepatology · doi:10.1002/hep.510280506 · Cited by 52 · see citing works
Bile canalicular dynamics in hepatocyte sandwich cultures
Reif, R., Karlsson, J., Günther, G., Beattie, L., Wrangborg, D., Hammad, S., Begher‐Tibbe, B., Damle‐Vartak, A., Melega, S., Kaye, P. M., et al. (2015) · Archives of Toxicology · doi:10.1007/s00204-015-1575-9 · Cited by 50 · see citing works
Hemery, I., Durand‐Schneider, A., Feldmann, G., Vaerman, J., & Maurice, M. (1996) · Journal of Cell Science · doi:10.1242/jcs.109.6.1215 · Cited by 45 · see citing works
Role of cytoskeleton in canalicular contraction in cultured differentiated hepatocytes.
Kawahara, H., & French, S. W. (1990) · PubMed · Cited by 37 · see citing works
Tsukada, N., & Phillips, M. J. (1993) · Journal of Histochemistry & Cytochemistry · doi:10.1177/41.3.7679126 · Cited by 30 · see citing works
Segawa, O., Miyano, T., Fujimoto, T., et al. (1993) · Journal of Pediatric Surgery · doi:10.1016/0022-3468(93)90344-k · Cited by 25 · see citing works
Torok, N. J., LaRusso, E. M., & McNiven, M. A. (2001) · Gastroenterology · doi:10.1053/gast.2001.28652 · Cited by 23 · see citing works
Coordinated Movement of Bile Canalicular Networks Reconstructed by Rat Small Hepatocytes
Sudo, R., Kohara, H., Mitaka, T., Ikeda, M., & Tanishita, K. (2005) · Annals of Biomedical Engineering · doi:10.1007/s10439-005-1690-5 · Cited by 20 · see citing works
Song, J., van Marle, J., Noorden, C. J. F. V., et al. (1996) · Histochemistry and Cell Biology · doi:10.1007/bf02473272 · Cited by 15 · see citing works
Larkin, J. M., Coleman, H. A., Espinosa, A., Levenson, A. E., Park, M. S., Woo, B., Zervoudakis, A., & Tinh, V. (2003) · Hepatology · doi:10.1053/jhep.2003.50419 · Cited by 13 · see citing works
Morphologic structure of bile canaliculi after bile duct ligation in the rat. A time-course study.
Vital, A., Bioulac‐Sage, P., Iron, A., et al. (1982) · PubMed · Cited by 13 · see citing works
Song, J., Noorden, C. J. F. V., & Frederiks, W. M. (1998) · Hepatology · doi:10.1002/hep.510270318 · Cited by 11 · see citing works
Initial site of bile regurgitation following extrahepatic biliary obstruction in living rats
Watanabe, N., Kojima, S., Takashimizu, S., Nishizaki, Y., Kagawa, T., & Phillips, M. J. (2006) · Journal of Gastroenterology and Hepatology · doi:10.1111/j.1440-1746.2006.04623.x · Cited by 11 · see citing works
Yokomori, H., Oda, M., Kamegaya, Y., Ogi, M., Tsukada, N., & Ishii, H. (2001) · Medical Molecular Morphology · doi:10.1007/s007950170005 · Cited by 10 · see citing works
Biliary hypertension as the cell proliferation trigger in bile duct ligated rats.
Azmaiparashvili, E., Kordzaia, D., & Dzidziguri, D. (2009) · PubMed · Cited by 8 · see citing works
Scanning electron microscopy on the rat liver with alphanaphthylisothiocyanate-induced cholestasis
Yoshino, K. (1980) · Gastroenterologia Japonica · doi:10.1007/bf02773758 · Cited by 6 · see citing works
Flagged first, then by citations
Research works printed but not linked: 2
- Matter A, Orci L, Rouiller C. A study on the permeability barriers between Disse's space and the bile canaliculus. Journal of ultrastructure research 1969;11:1-71.
- Seglen PO. Preparation of isolated rat liver cells. Methods in cell biology 1976;13:29-83.
Cited by
Sorted by citations
Zhao, Z., Chen, X., Dowbaj, A. M., Sljukic, A., Bratlie, K., Lin, L., Fong, E. L. S., Balachander, G. M., Chen, Z., Soragni, A., et al. (2022) · Nature Reviews Methods Primers · doi:10.1038/s43586-022-00174-y · Cited by 1,001 · see citing works
Self-citationMouse liver assembloids model periportal architecture and biliary fibrosis
Dowbaj, A. M., Sljukic, A., Niksic, A., Landerer, C., Delpierre, J., Yang, H., Lahree, A., Kühn, A. C., Beers, D., Byrne, H. M., et al. (2025) · Nature · doi:10.1038/s41586-025-09183-9 · Cited by 40 · see citing works
Gupta, K., Ng, I. C., Balachander, G. M., Nguyen, B. P., Tucker-Kellogg, L., Low, B. C., & Yu, H. (2020) · Biomaterials · doi:10.1016/j.biomaterials.2020.120283 · Cited by 23 · see citing works
Self-citation
- Authors
- Gupta, K., Li, Q., Fan, J. J., Fong, E. L. S., Song, Z., Mo, S., Tang, H., Ng, I. C., Ng, C. W., Pawijit, P., Zhuo, S., Dong, C.-Y., Low, B. C., Wee, A., Dan, Y. Y., Kanchanawong, P., So, P., Viasnoff, V., & Yu, H.
- Affiliations
- National University of Singapore (SG)
- Agency for Science, Technology and Research (SG)
- Fujian Normal University (CN)
- National Taiwan University (TW)
- National University Hospital (SG)
- Singapore-MIT Alliance for Research and Technology (SG)
- Centre National de la Recherche Scientifique (FR)
- Corresponding author
- Yu, Hanry
- ORCID
- Venue
- Journal of Hepatology
- Publisher
- Elsevier BV
- Published
- 2017-06
- Volume
- 66
- Issue
- 6
- Pages
- 1231-1240
- ISSN
- 0168-8278 (ISSN-L)
- OpenAlex
- W2586247078
- Cited by
- 29
- References
- 53
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 9 · 2023: 5 · 2022: 2 · 2021: 2 · 2020: 3 · 2018: 5 · 2017: 1
- FWCI
- 1.52
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Locations
- Journal of Hepatology (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- National University of Singapore (submittedVersion)
- Funders
- National University of Singapore
- Mechanobiology Institute, Singapore
- National Institute of Biomedical Imaging and Bioengineering (P41EB015871)
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Health Sciences › Medicine › Hepatology › Liver Diseases and Immunity
- Keywords
- actomyosin contractility, obstructive cholestasis, bile canaliculi, biliary obstruction, actin polymerization, bile duct ligation
- MeSH terms
- Actomyosin, Animals, Bile Canaliculi, Bile Ducts, Bile Reflux, Biomechanical Phenomena, Cholestasis, Disease Models, Animal, Male, Mice, Transgenic, Pressure, Rats, Wistar, Mice, Rats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
