title: Dual labeling with 5-bromo-2'-deoxyuridine and 5-ethynyl-2'-deoxyuridine for estimation of cell migration rate in the small intestinal epithelium.
abstract: Small intestinal epithelium is a self-renewing system in which the entire sequence of cell proliferation, differentiation, and removal is coupled to cell migration along the crypt-villus axis. We examined whether dual labeling with different thymidine analogues, 5-bromo-2'-deoxyuridine (BrdU) and 5-ethynyl-2'-deoxyuridine (EdU), can be used to estimate cell migration rates on the villi of small intestines in rats. Rats received a single intraperitoneal injection of BrdU and EdU within a time interval, and signals in tissue sections were examined by immunohistochemistry and the "click" reaction, respectively. We successfully observed BrdU- and EdU-positive cells on the epithelium with no cross-reaction. In addition, we observed an almost complete overlapping of BrdU- and EdU-positive cells in rats administered simultaneously with BrdU and EdU. By calculating the cell migration rate by dividing the distance between the median cell positions of the distribution of BrdU- and EdU-positive cells by the time between the injection of BrdU and EdU, we estimated approximately 9 and 5 μm/h for the cell migration rates on the villi in the jejunum and ileum, respectively. We propose that dual labeling with BrdU and EdU within a time interval, followed by detecting with immunohistochemistry and the click reaction, respectively, is useful to estimate accurately the cell migration rate in the intestinal epithelium in a single animal.
authors.affiliation: Graduate School of Life Science, Hokkaido University, Sapporo, 060-8589, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Asano
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tsuruta
authors.forename: Naomichi
authors.initials: N
authors.lastname: Nishimura
authors.forename: Kei
authors.initials: K
authors.lastname: Sonoyama
mesh_terms: D000818:Animals; D001973:Bromodeoxyuridine / Q000494:pharmacology*; D002465:Cell Movement / Q000502:physiology*; D003857:Deoxyuridine / Q000031:analogs & derivatives* / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D001973:Bromodeoxyuridine; C031086:5-ethynyl-2'-deoxyuridine; D003857:Deoxyuridine
keywords: 5-bromo-2'-deoxyuridine; 5-ehynyl-2'-deoxyuridine; cell migration; intestine