Characterization of phagosomal subpopulations along endocytic routes in osteoclasts and macrophages

Eiko Sakai, Hisatsugu Miyamoto, Kuniaki Okamoto, Yuzo Kato, Kenji Yamamoto, Hideaki Sakai

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Modifications occurring during the transformation of phagosomes into mature phagolysosomes were investigated in osteoclast-like cells (OCLs) and macrophages using latex beads as markers for the isolation of phagosomal compartments (LBC) at different time points after phagocytosis. In OCLs, newly formed LBC acquired cathepsin K, tartarateresistant phosphatase (TRAP), lysosome-associated membrane protein-1 (Lamp-1), and cathepsin D, and rapidly lost annexin II in a time-dependent manner. The levels of Rab7 and c-Src in OCLs initially increased and then gradually decreased during the transformation from early to late endosomal LBC or phagolysosomes. Receptor activator of NF-κB (RANKL) significantly increased the LBC levels of cathepsin K, TRAP, and c-Src, whereas calcitonin decreased the LBC levels of cathepsin K, TRAP, and Rab7, indicating that the transformation of early to late endosomal elements and lysosomes in OCLs is also regulated by osteoclastogenesis regulatory factors. On the other hand, changes in the LBC levels of Lamp-1, cathepsin D, and annexin IT in macrophages were comparable to those in OCLs. However, contrary to osteoclastic LBC, Rab7 levels of macrophage LBC decreased in a time-dependent manner. Macrophage LBC were devoid of cathepsin K, TRAP, and c-Src in all transformation stages. These observations suggest that OCLs and macrophages have different phagosome maturation mechanisms that involve the specific and regulated acquisition of markers from endocytic organelles. The results also demonstrate that the use of LBC is a useful system in which to identify and characterize molecules involved in these different endocytic pathways.

Original languageEnglish
Pages (from-to)823-831
Number of pages9
JournalJournal of Biochemistry
Volume130
Issue number6
DOIs
Publication statusPublished - Dec 1 2001
Externally publishedYes

Fingerprint

Macrophages
Osteoclasts
Cathepsin K
Phagosomes
Phosphoric Monoester Hydrolases
Lysosome-Associated Membrane Glycoproteins
Cathepsin D
Annexin A2
Annexins
Latex
Calcitonin
Lysosomes
Microspheres
Phagocytosis
Osteogenesis
Organelles
Molecules

Keywords

  • Endocytosis
  • Macrophage
  • Osteoclast
  • Phagosome

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry
  • Molecular Biology

Cite this

Characterization of phagosomal subpopulations along endocytic routes in osteoclasts and macrophages. / Sakai, Eiko; Miyamoto, Hisatsugu; Okamoto, Kuniaki; Kato, Yuzo; Yamamoto, Kenji; Sakai, Hideaki.

In: Journal of Biochemistry, Vol. 130, No. 6, 01.12.2001, p. 823-831.

Research output: Contribution to journalArticle

Sakai, Eiko ; Miyamoto, Hisatsugu ; Okamoto, Kuniaki ; Kato, Yuzo ; Yamamoto, Kenji ; Sakai, Hideaki. / Characterization of phagosomal subpopulations along endocytic routes in osteoclasts and macrophages. In: Journal of Biochemistry. 2001 ; Vol. 130, No. 6. pp. 823-831.
@article{eab9348122ec4499866890c53639a78b,
title = "Characterization of phagosomal subpopulations along endocytic routes in osteoclasts and macrophages",
abstract = "Modifications occurring during the transformation of phagosomes into mature phagolysosomes were investigated in osteoclast-like cells (OCLs) and macrophages using latex beads as markers for the isolation of phagosomal compartments (LBC) at different time points after phagocytosis. In OCLs, newly formed LBC acquired cathepsin K, tartarateresistant phosphatase (TRAP), lysosome-associated membrane protein-1 (Lamp-1), and cathepsin D, and rapidly lost annexin II in a time-dependent manner. The levels of Rab7 and c-Src in OCLs initially increased and then gradually decreased during the transformation from early to late endosomal LBC or phagolysosomes. Receptor activator of NF-κB (RANKL) significantly increased the LBC levels of cathepsin K, TRAP, and c-Src, whereas calcitonin decreased the LBC levels of cathepsin K, TRAP, and Rab7, indicating that the transformation of early to late endosomal elements and lysosomes in OCLs is also regulated by osteoclastogenesis regulatory factors. On the other hand, changes in the LBC levels of Lamp-1, cathepsin D, and annexin IT in macrophages were comparable to those in OCLs. However, contrary to osteoclastic LBC, Rab7 levels of macrophage LBC decreased in a time-dependent manner. Macrophage LBC were devoid of cathepsin K, TRAP, and c-Src in all transformation stages. These observations suggest that OCLs and macrophages have different phagosome maturation mechanisms that involve the specific and regulated acquisition of markers from endocytic organelles. The results also demonstrate that the use of LBC is a useful system in which to identify and characterize molecules involved in these different endocytic pathways.",
keywords = "Endocytosis, Macrophage, Osteoclast, Phagosome",
author = "Eiko Sakai and Hisatsugu Miyamoto and Kuniaki Okamoto and Yuzo Kato and Kenji Yamamoto and Hideaki Sakai",
year = "2001",
month = "12",
day = "1",
doi = "10.1093/oxfordjournals.jbchem.a003054",
language = "English",
volume = "130",
pages = "823--831",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "6",

}

TY - JOUR

T1 - Characterization of phagosomal subpopulations along endocytic routes in osteoclasts and macrophages

AU - Sakai, Eiko

AU - Miyamoto, Hisatsugu

AU - Okamoto, Kuniaki

AU - Kato, Yuzo

AU - Yamamoto, Kenji

AU - Sakai, Hideaki

PY - 2001/12/1

Y1 - 2001/12/1

N2 - Modifications occurring during the transformation of phagosomes into mature phagolysosomes were investigated in osteoclast-like cells (OCLs) and macrophages using latex beads as markers for the isolation of phagosomal compartments (LBC) at different time points after phagocytosis. In OCLs, newly formed LBC acquired cathepsin K, tartarateresistant phosphatase (TRAP), lysosome-associated membrane protein-1 (Lamp-1), and cathepsin D, and rapidly lost annexin II in a time-dependent manner. The levels of Rab7 and c-Src in OCLs initially increased and then gradually decreased during the transformation from early to late endosomal LBC or phagolysosomes. Receptor activator of NF-κB (RANKL) significantly increased the LBC levels of cathepsin K, TRAP, and c-Src, whereas calcitonin decreased the LBC levels of cathepsin K, TRAP, and Rab7, indicating that the transformation of early to late endosomal elements and lysosomes in OCLs is also regulated by osteoclastogenesis regulatory factors. On the other hand, changes in the LBC levels of Lamp-1, cathepsin D, and annexin IT in macrophages were comparable to those in OCLs. However, contrary to osteoclastic LBC, Rab7 levels of macrophage LBC decreased in a time-dependent manner. Macrophage LBC were devoid of cathepsin K, TRAP, and c-Src in all transformation stages. These observations suggest that OCLs and macrophages have different phagosome maturation mechanisms that involve the specific and regulated acquisition of markers from endocytic organelles. The results also demonstrate that the use of LBC is a useful system in which to identify and characterize molecules involved in these different endocytic pathways.

AB - Modifications occurring during the transformation of phagosomes into mature phagolysosomes were investigated in osteoclast-like cells (OCLs) and macrophages using latex beads as markers for the isolation of phagosomal compartments (LBC) at different time points after phagocytosis. In OCLs, newly formed LBC acquired cathepsin K, tartarateresistant phosphatase (TRAP), lysosome-associated membrane protein-1 (Lamp-1), and cathepsin D, and rapidly lost annexin II in a time-dependent manner. The levels of Rab7 and c-Src in OCLs initially increased and then gradually decreased during the transformation from early to late endosomal LBC or phagolysosomes. Receptor activator of NF-κB (RANKL) significantly increased the LBC levels of cathepsin K, TRAP, and c-Src, whereas calcitonin decreased the LBC levels of cathepsin K, TRAP, and Rab7, indicating that the transformation of early to late endosomal elements and lysosomes in OCLs is also regulated by osteoclastogenesis regulatory factors. On the other hand, changes in the LBC levels of Lamp-1, cathepsin D, and annexin IT in macrophages were comparable to those in OCLs. However, contrary to osteoclastic LBC, Rab7 levels of macrophage LBC decreased in a time-dependent manner. Macrophage LBC were devoid of cathepsin K, TRAP, and c-Src in all transformation stages. These observations suggest that OCLs and macrophages have different phagosome maturation mechanisms that involve the specific and regulated acquisition of markers from endocytic organelles. The results also demonstrate that the use of LBC is a useful system in which to identify and characterize molecules involved in these different endocytic pathways.

KW - Endocytosis

KW - Macrophage

KW - Osteoclast

KW - Phagosome

UR - http://www.scopus.com/inward/record.url?scp=0035667183&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035667183&partnerID=8YFLogxK

U2 - 10.1093/oxfordjournals.jbchem.a003054

DO - 10.1093/oxfordjournals.jbchem.a003054

M3 - Article

VL - 130

SP - 823

EP - 831

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 6

ER -