Nuclearity and BrdU Labeling of Rat Hepatocytes in Cytocentrifuge Preparations of Freshly Isolated Hepatocytes with Cumulative Labeling of Bromodeoxyuridine

Fujii, Etsuko; Karasawa, Yayoi; Kumano, Eiichi; Sakurai, Takayuki; Misawa, Yasuyuki; Mori, Toshio; Ito, Tsuneo; Suzuki, Masami; Sugimoto, Tetsuro

doi:10.1293/tox.17.43

Cited by 4 publications

(11 citation statements)

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“…This means that after one, two, and four weeks of cumulative labeling, positive cells have gone through the S phase during these periods. WP cells have two nuclei that have gone through the S phase, confirming that binuclear hepatocytes are mainly formed by acytokinetic mitosis (Fujii et al 2004). Similarly, in the present study, WP binuclear cells were dominant, indicating that at least a part of the binuclear cells were formed by acytokinetic mitosis.…”

Section: Discussionsupporting

confidence: 86%

“…BrdU labeling was carried out according to the method described by Fujii et al (2004). Briefly, rats kept under ether anesthesia were implanted intraperitoneally with mini-osmotic pumps (Alzet Model 2002, Alza Co., Palo Alto, CA, USA) that were filled with 12% BrdU (Sigma Chemical Co., St. Louis, MO, USA) in distilled water containing 50% dimethyl sulfoxide (Sigma Chemical Co., St. Louis, MO, USA) and released at 240 μg of BrdU per hour for twenty-eight days.…”

Section: Experiments 2: Cell Kinetic Analysis Of Mnhs Induced By Mitemcinal In Ratsmentioning

confidence: 99%

“…In addition, we have applied a cell kinetic INTRODUCTION Mononuclear and binuclear hepatocytes are commonly observed in the liver. The binuclear cell population is approximately 30%-40% in normal rat liver (Engelmann et al 1981;Fujii et al 2004;Gerlyng et al 1993;Mossin et al 1994;Seglen 1997;Wheatley 1972). Although the mechanisms are unclear, the ratio of mono-and binuclear hepatocytes is known to change with age (Engelmann et al 1981;Gerlyng et al 1993;Seglen 1997;Wheatley 1972), partial hepatectomy (Auvigne et al 2002;Frederiks et al 1990;Gerlyng et al 1993;Melchiorri et al 1993;Seglen 1997;Wheatley 1972), endocrine status (Lamers 1985;Mossin et al 1994;Wheatley 1972), and certain hepatomitogenic and hepatocarcinogenic drugs (Auvigne et al 2002;Gerlyng et al 1994;Kostka et al 1994;Kostka et al 2000;Melchiorri et al 1993;Palut et al 1992;Seglen 1997).…”

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Characterization of Multinuclear Hepatocytes Induced in Rats by Mitemcinal (GM-611), an Erythromycin Derivative

Hayashi¹,

Fujii²,

Kato³

et al. 2008

Toxicol Pathol

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Mitemcinal is an erythromycin derivative with motilin agonistic action, developed as a gastrointestinal motor-activating agent. The characteristics of mitemcinal-induced multinuclear hepatocytes (MNHs, hepatocytes with three or more nuclei per cell) from detailed morphological observations together with the results of a study on the mechanisms of MNH formation by combining cytocentrifuge preparations with 5-bromo-2'-deoxyuridine cumulative labeling are reported. MNHs were observed only in rats in the high-dose groups of the subchronic study, with a higher incidence in females and reversibility after twenty-eight days of drug withdrawal, but not observed in dogs. In the chronic study, the incidence increased relative to the dose. Histopathologically, MNHs were preferentially observed in the centrilobular zone, without nuclear atypia or mitotic figures. In the cell kinetic study, the labeling pattern of MNHs included all-positive, all-negative, and mixed labeling patterns of nuclei. The all-negative pattern indicated that the cells were formed by fusion of nondividing cells. The current results indicate that the cell kinetic approach effectively demonstrated the mechanism of mitemcinal-induced MNHs as fusion of hepatocytes and that drug-induced disturbance of mitosis is not involved in the multinucleation of MNHs by mitemcinal.

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Section: Discussionsupporting

confidence: 86%

Section: Experiments 2: Cell Kinetic Analysis Of Mnhs Induced By Mitemcinal In Ratsmentioning

confidence: 99%

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confidence: 99%

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Characterization of Multinuclear Hepatocytes Induced in Rats by Mitemcinal (GM-611), an Erythromycin Derivative

Hayashi¹,

Fujii²,

Kato³

et al. 2008

Toxicol Pathol

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“…In normal rat liver, binucleate cells are quite common and can make up approximately 30% of the total hepatocyte population (Seglen, 1997;Fuji et al, 2004) in vivo. A small number of multinucleated cells (greater than two nuclei) have been reported as well; however, an increased occurrence of multinucleate cells has been associated with a number of diseases (Lu and Kang, 2009) and certain drugs (Scampini et al, 1993;Seglen 1997;Kostka et al, 2000;Auvigne et al, 2002;Hayashi et al, 2008).…”

Section: Commentary Background Informationmentioning

confidence: 99%

Assessing Cell Fusion and Cytokinesis Failure as Mechanisms of Clone 9 Hepatocyte Multinucleation In Vitro

et al. 2012

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In this in vitro model of hepatocyte multinucleation, separate cultures of rat Clone 9 cells are labeled with either red or green cell tracker dyes (Red Cell Tracker CMPTX or Vybrant CFDA SE Cell Tracer), plated together in mixed-color colonies, and treated with positive or negative control agents for 4 days. The fluorescent dyes become cell-impermeant after entering cells and are not transferred to adjacent cells in a population, but are inherited by daughter cells after fusion. The mixed-color cultures are then evaluated microscopically for multinucleation and analysis of the underlying mechanism (cell fusion/cytokinesis). Multinucleated cells containing only one dye have undergone cytokinesis failure, whereas dual-labeled multinucleated cells have resulted from fusion.

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“…BrdU labeling was carried out according to the method described by Suzuki et al (1998) and Fujii et al (2004). For flash labeling, the rats received an intravenous injection of an aqueous solution of BrdU and 5-fluoro-2'-deoxyuridine (10:1 ratio; Amersham, Bucks, UK) in the tail vein at a dose level of 30 mg/kg and were sacrificed 2 hr after the injection.…”

Section: Brdu Labelingmentioning

confidence: 99%

Cell proliferative activity in the kidney of young growing rat analyzed using flash and cumulative labeling with bromodeoxyuridine

et al. 2010

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-We examined the cell proliferation activity of kidney in young growing rats using flash and cumulative labeling with bromodeoxyuridine (BrdU). Rats were subjected to the study at the age of 6 weeks, and cumulative labeling was carried out for periods of 7 to 28 days. BrdU-positive cells were observed after flash labeling and were increased by cumulative labeling. The positive epithelia were mainly distributed in the cortex and the outer stripe of the outer medulla and were scarce in the inner stripe of the outer medulla and inner medulla throughout all labeling periods. In the tubular epithelium, the majority of positive cells were found in the proximal tubule. In the proximal tubule, positive epithelia were abundant in the medullary rays and in the outer stripe of the outer medulla. In the intermediate tubule to collecting duct, positive epithelia were rare. In the renal corpuscle, positive nuclei were mainly found in the endothelial cells and the mesangial cells and were scarce in the parietal cells of the Bowman's capsule. BrdU-positive nuclei were not observed in podocytes. These results indicate that renal tubules actively grow relative to epithelial proliferation, and that the endothelial cells, the mesangial cells and the parietal cells in the renal corpuscle also proliferate at the age of 6 to 10 weeks in rats. For assessment of renal toxicity using young growing rats, not only the morphologic and physiologic features unique to the kidney but also the growing process of the kidney should be taken into account.

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Nuclearity and BrdU Labeling of Rat Hepatocytes in Cytocentrifuge Preparations of Freshly Isolated Hepatocytes with Cumulative Labeling of Bromodeoxyuridine

Cited by 4 publications

References 20 publications

Characterization of Multinuclear Hepatocytes Induced in Rats by Mitemcinal (GM-611), an Erythromycin Derivative

Characterization of Multinuclear Hepatocytes Induced in Rats by Mitemcinal (GM-611), an Erythromycin Derivative

Assessing Cell Fusion and Cytokinesis Failure as Mechanisms of Clone 9 Hepatocyte Multinucleation In Vitro

Cell proliferative activity in the kidney of young growing rat analyzed using flash and cumulative labeling with bromodeoxyuridine

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