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Abnormal expression of Muc5b in Cftr-null mice and in mammary tumors of MMTV-ras mice

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Abstract

Gel-forming mucins are large, high molecular weight, and heavily O-glycosylated proteins that are responsible for the rheological properties of mucus gel. Among them, the mucin MUC5B has been implicated in breast cancer and cystic fibrosis. We obtained a new polyclonal serum, named CP1, which was isolated from a rabbit immunized with a mouse Muc5b peptide. The immunoprofile of Muc5b was determined on paraffin-embedded and frozen mouse tissue sections and showed a similar expression pattern in mouse to that in the human. The “nonmammary” mucin Muc5b was detected in all mammary tumors analyzed from MMTV-ras mice, suggesting that the CP1 antibody is a valuable tool for investigating the involvement of this mucin in mammary cancer. We also found that uninfected Cftr −/− mice harbored more Clara cells, which were Muc5b-positive, than did their wild-type control littermates. The number of Muc5b-positive cells increased in Cftr −/− mice infected experimentally with Pseudomonas aeruginosa, and the mice developed mucus plugs in their bronchi and bronchioles with a high frequency of Muc5b content (87%, Cohen’s kappa = 0.82; p < 0.0001). These findings suggest that mice genetically deficient in the Cftr gene are predisposed to develop mucus plugs and that MUC5B may provide a valuable target for decreasing mucus viscosity in cystic fibrosis.

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Acknowledgments

We thank the microbiology department of the Centre de Biologie-Pathologie Pierre-Marie Degand (Lille Hospital) for access to the ABI Prism 7700; MH Gevaert and RM Siminski (Service Commun-Morphologie cellulaire, Université Lille 2) for slides; J Bara (Inserm UMR S938, Paris, France) for the 45M1 antibody; J Ryerse (Department of Pathology, Saint Louis, MO) for the anti-CCSP (R42AP) antibody; and MR Parsek and BR Borlee (Seattle, WA) for the PAO1 strain. This study was supported by the French Cystic Fibrosis Association Vaincre la Mucoviscidose (JLD, FC0203, and IC0806).

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Authors and Affiliations

  1. Inserm U995, Faculté de Médecine, Pôle Recherche, Bât. Huriez, 5ème étage, University Lille 2, 59045, Lille, France

    Hélène Valque, Valérie Gouyer, Marie-Odile Husson, Frédéric Gottrand & Jean-Luc Desseyn

  2. CHRU of Lille, Lille, France

    Valérie Gouyer, Marie-Odile Husson & Frédéric Gottrand

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  1. Hélène Valque
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  2. Valérie Gouyer
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  3. Marie-Odile Husson
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  4. Frédéric Gottrand
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  5. Jean-Luc Desseyn
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Correspondence to Jean-Luc Desseyn.

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418_2011_872_MOESM1_ESM.tif

Supplementary Figure 1. Location of epitopes used for anti-mouse Muc5b antibody production. A Schematic representation of the mouse mucin Muc5b. B The known CYS protein sequences of mouse-secreted gel-forming mucins were aligned to show sequence similarity. The peptide used for antibody production and shorter peptides that may also be immunogens are highlighted in gray. Until now, only one CYS domain and five CYS domains of Muc5ac have been sequenced; we named the first three (CYS#1–3) and the last two CYS#8 and #9 of Muc5ac by analogy to the nine CYS domains of the human counterparts. However, we assumed that mouse Muc2 and Muc5ac contain two and nine CYS domains, respectively, as do their human orthologues

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Valque, H., Gouyer, V., Husson, MO. et al. Abnormal expression of Muc5b in Cftr-null mice and in mammary tumors of MMTV-ras mice. Histochem Cell Biol 136, 699–708 (2011). https://doi.org/10.1007/s00418-011-0872-5

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