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Altered expression of tight junction proteins in mammary epithelium after discontinued suckling in mice

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Abstract

Milk production is modulated by the paracellular barrier function of tight junction (TJ) proteins located in the mammary epithelium. The aim of our study was the molecular analysis of TJs in native lactating murine mammary gland epithelium as this process may strongly challenge epithelial barrier properties and regulation. Mammary gland tissue specimens from lactating control mice and animals after a 20-h interruption of suckling were prepared; histological analyses were performed by light and electron microscopy; and expression of TJ proteins was detected by PCR, Western blotting, immunofluorescent staining, and confocal laser scanning microscopy. Discontinuation of suckling resulted in a substantial accumulation of milk in mammary glands, an increase of alveolar size, and a flattening of epithelial cells without effects on inflammatory indicators. In control tissues, PCR and Western blots showed signals for occludin, and claudin-1, -2, -3, -4, -5, -7, -8, -15, and -16. After a 20-h accumulation of milk, expression of two sealing TJ proteins, claudin-1 and -3, was markedly increased, whereas two TJ proteins involved in cation transport, claudin-2 and -16, were reduced. Real-time PCR validated increased transcripts of claudin-1 and claudin-3. During extension of mammary glands in the process of lactation, claudin-1 and -3 are markedly induced and claudin-2 and -16 are decreased. Volume and composition of milk might be strongly dependent on this counter-regulation of sealing claudins with permeability-mediating claudins, indicating a physiological process of a tightening of TJs against a back-leak of solutes and ions from the alveolar lumen.

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Abbreviations

TJ:

Tight junction

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Acknowledgments

The study has been supported by the Deutsche Forschungsgemeinschaft (DFG FOR 721), the Sonnenfeld-Stiftung Berlin, the Partnership Program FU Berlin–University St. Petersburg, and by Saint Petersburg University Research Grant No. 1.37.118.2011.

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There is no conflict of interest.

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Authors and Affiliations

  1. Biological and Soil Faculty, St. Petersburg University, Universitetskaya nab. 7/9, 199034, St. Petersburg, Russia

    Alexander G. Markov, Natalia M. Kruglova & Yulia A. Fomina

  2. Institute of Clinical Physiology, Charité Campus Benjamin Franklin, 12200, Berlin, Germany

    Michael Fromm & Salah Amasheh

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  1. Alexander G. Markov
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  2. Natalia M. Kruglova
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  3. Yulia A. Fomina
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  4. Michael Fromm
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Correspondence to Salah Amasheh.

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Markov, A.G., Kruglova, N.M., Fomina, Y.A. et al. Altered expression of tight junction proteins in mammary epithelium after discontinued suckling in mice. Pflugers Arch - Eur J Physiol 463, 391–398 (2012). https://doi.org/10.1007/s00424-011-1034-2

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  • DOI: https://doi.org/10.1007/s00424-011-1034-2

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