Immunocytochemical identification of proliferating cell types in mouse mammary gland

A. Sapino, L. Macri, P. Gugliotta, G. Bussolati

Research output: Contribution to journalArticlepeer-review

Abstract

To study cell proliferation in different cell types and segments of the mammary gland, we devised a dual staining procedure, combining nuclear labeling by 5-bromo-2'-deoxyuridine (BrdU) uptake (revealed by a dark-brown precipitate) and an alternative (red or blue) cytoplasmic labeling by antibodies specific for the differentiation proteins of epithelial, myoepithelial, and secretory cell types. The following markers, revealed by APAAP or β-galactosidase procedure, were selected: α-smooth muscle actin for the myoepithelial cells, keratin (detected by AE1 monoclonal) for the luminal epithelial cells, α-lactalbumin and β-casein for the secretory cells. To follow the full process of organogenesis, the study was conducted in mouse mammary glands from virgin, primed, and lactating animals and from glands cultured in vitro under specific hormone stimulation. Cell proliferation was localized mainly in focal areas (end buds), and mostly corresponded to 'null' undifferentiated cells. Estrogen and progestin stimulation induced a relative increase of proliferating differentiated cells of either epithelial or myoepithelial type, localized in ducts and alveolar structures. Prolactin stimulation induced proliferation in secretory cells.

Original languageEnglish
Pages (from-to)1541-1547
Number of pages7
JournalJournal of Histochemistry and Cytochemistry
Volume38
Issue number11
Publication statusPublished - 1990

Keywords

  • 5-Bromo-2'-deoxyuridine
  • Cell differentiation
  • Immunocytochemistry
  • Mammary gland
  • Proliferation

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

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