CM2 antigen, a potential novel molecule participating in glucuronide transport on rat hepatocyte canalicular membrane

Submitted: 4 December 2011
Accepted: 19 March 2012
Published: 29 June 2012
Abstract Views: 755
PDF: 405
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Authors

The polarized molecules predominately distributing at hepatocyte canalicular surface play a vital role in disclosing the process of bile formation and etiopathogenisis of cholestatic live diseases. Therefore, it is important to find novel polarized molecules on hepatocyte canalicular membrane. In the present study, canalicular membrane vesicles (CMVs) isolated from rat hepatocyte by density gradient centrifugation were used as immunogens to produce hybridoma and 46 strains of monoclonal antibodies (mAb) against CMVs were obtained. With a series of morphological assay methods, including immunohistochemistry, immunofluorescence and immuno-electron microscope, the antigens recognized by canalicular mAb1 (CM1) and canalicular mAb2 (CM2) were confirmed to predominately distribute at hepatocyte canalicular membrane. Transport activity assay revealed that CM2 could inhibit ATP-dependent E217βG uptake of rat hepatocyte CMVs. Meanwhile, Western blotting analysis showed that the molecular mass of CM2 antigen was approximately 110kDa, which was much less than Mr 180kDa of multidrug resistance-associated protein 2 (MRP2) involved in glucuronide transport. These data indicated that CM2 antigen might be a potential novel molecule participating in glucuronide transport on the hepatocyte canalicular membrane.

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Supporting Agencies

this study was supported by the National Natural Science Foundation of China (NO. 30971339, 30971116, 81071873 and 81070326).
J. Wang, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
X. Zhou, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
J. Li, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
Y. Shi, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
Z. Han, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
X. Wang, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
S. Li, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
Z. Yang, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
R. Wang, The Fourth Military Medical University
Department of Pathology
D. Fan, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases
Y. Han, The Fourth Military Medical University
Division of Hepatology, Xijing Hospital of Digestive Diseases

How to Cite

Wang, L., Wang, J., Zhou, X., Li, J., Shi, Y., Han, Z., Wang, X., Li, S., Yang, Z., Wang, R., Fan, D., & Han, Y. (2012). CM2 antigen, a potential novel molecule participating in glucuronide transport on rat hepatocyte canalicular membrane. European Journal of Histochemistry, 56(3), e26. https://doi.org/10.4081/ejh.2012.e26