title: Accurate determination of S-phase fraction in proliferative cells by dual fluorescence and peroxidase immunohistochemistry with 5-bromo-2'-deoxyuridine (BrdU) and Ki67 antibodies.
abstract: To ensure the maintenance of tissues in mammals, cell loss must be balanced with cell production, the proliferative activity being different from tissue to tissue. In this article, the authors propose a new method for the quantification of the proliferative activity, defined as the S-phase fraction of actively cycling cells, by dual labeling with fluorescence and peroxidase immunohistochemistry using BrdU (marker of S-phase) and Ki67 antibodies (marker of G(1)-, S-, G(2)-, and M-phases) after a one-step antigen retrieval. In the generative cell zones of fundic and pyloric glandular stomachs, where the majority of cells were cycling, the authors measured a proliferative activity of 31%. In the epithelium of the forestomach and the skin, where cycling cells are intermingled with G(0) and differentiated cells, proliferative activities were 21% and 13%, respectively. In the adrenal cortex, in which cycling cells were sparsely distributed, the proliferative activity reached 32%. During the regenerative process in the skin after a lesion, the proliferative activity increased in proximity to the wound. The present one-step dual-labeling method has revealed that the proliferative activity is different between tissues and depends on the physiological or pathological state.
authors.affiliation: Department of Systems Biology, Graduate School of Pharmacological Science, Kyoto University, Kyoto, Japan.
authors.forename: Rina
authors.initials: R
authors.lastname: Tanaka
authors.forename: Motomi
authors.initials: M
authors.lastname: Tainaka
authors.forename: Takumi
authors.initials: T
authors.lastname: Ota
authors.forename: Naoki
authors.initials: N
authors.lastname: Mizuguchi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kato
authors.forename: Shoichi
authors.initials: S
authors.lastname: Urabe
authors.forename: Yulin
authors.initials: Y
authors.lastname: Chen
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Fustin
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Yamaguchi
authors.forename: Masao
authors.initials: M
authors.lastname: Doi
authors.forename: Shinshichi
authors.initials: S
authors.lastname: Hamada
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Okamura
mesh_terms: D000311:Adrenal Glands / Q000166:cytology / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D001973:Bromodeoxyuridine; D049109:Cell Proliferation; D005753:Gastric Mucosa / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D019394:Ki-67 Antigen / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016196:S Phase; D012867:Skin / Q000166:cytology / Q000293:injuries / Q000378:metabolism; D013194:Staining and Labeling; D013270:Stomach / Q000166:cytology; D014945:Wound Healing
chemical_list: D000906:Antibodies; D007202:Indicators and Reagents; D019394:Ki-67 Antigen; D001973:Bromodeoxyuridine