Detection and localization of cripto-1 binding in mouse mammary epithelial cells and in the mouse mammary gland using an immunoglobulin - Cripto-1 fusion protein

Caterina Bianco, Nicola Normanno, Antonella De Luca, Monica Rosaria Maiello, Christian Wechselberger, Youping Sun, Nadia Khan, Heather Adkins, Michele Sanicola, Barbara Vonderhaar, Bruce Cohen, Masaharu Seno, David Salomon

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18 Citations (Scopus)

Abstract

Human Cripto-1 (CR-1), a member of the epidermal growth factor-CFC (EGF-CFC) family of peptides, is expressed in the developing mouse mammary gland and can modulate mammary epithelial cell migration, branching morphogenesis and milk protein expression in vitro. In order to screen for a CR-1 receptor and to identify potential CR-1 target tissues, we constructed a fusion protein comprising the EGF-like domain of CR-1 and the Fc domain of a human IgG1. The recombinant CR-1 fusion protein (CR-1-Fc) was biologically active as it was able to activate the ras/raf/mitogen activated protein kinase (MAPK) pathway and to inhibit transcription of the milk protein β-casein in NMuMG and HC-11 mouse mammary epithelial cells. By using immunocytochemistry and by an in situ enzyme-linked immunosorbent assay (ELISA), CR-1-Fc was found to specifically bind to NMuMG and HC-11 cells. Finally, immunohistochemical analysis using CR-1-Fc showed a specific localization of CR-1 binding to tissue sections from mouse mammary gland. In particular, more than 60% of the epithelial cells were intensely stained with the CR-1-Fc fusion protein in the lactating mouse mammary gland, whereas approximately 25% of the mammary epithelial cells were stained in the gland from pregnant mouse. Since expression of mouse cripto-1 (Ct-1) in the pregnant and lactating mouse mammary gland as well as its presence in milk has been previously demonstrated, these data strongly suggest that an autocrine pathway involving Cr-1 and its putative receptor is operating in the mouse mammary gland during pregnancy and lactation.

Original languageEnglish
Pages (from-to)74-82
Number of pages9
JournalJournal of cellular physiology
Volume190
Issue number1
DOIs
Publication statusPublished - Jan 1 2002
Externally publishedYes

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

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    Bianco, C., Normanno, N., De Luca, A., Maiello, M. R., Wechselberger, C., Sun, Y., Khan, N., Adkins, H., Sanicola, M., Vonderhaar, B., Cohen, B., Seno, M., & Salomon, D. (2002). Detection and localization of cripto-1 binding in mouse mammary epithelial cells and in the mouse mammary gland using an immunoglobulin - Cripto-1 fusion protein. Journal of cellular physiology, 190(1), 74-82. https://doi.org/10.1002/jcp.10037