Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites.

Bravo, Rodrigo; Macdonald-Bravo, H

doi:10.1083/jcb.105.4.1549

Cited by 875 publications

(506 citation statements)

References 31 publications

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“…By detergent treatment, the main bulk of PCNA is extracted, whereas the S-phase associated form is resistant (1). The cell cycle expression of PCNA using the human autoantibody was unchanged in unfixed nuclei, indicating that the reactivity in alcohol fixed cells indeed is S-phase specific.…”

Section: Discussionmentioning

confidence: 98%

Flow cytometric analysis of proliferation associated nuclear antigens using washless staining of unfixed cells

Landberg

Roos

1992

Cytometry

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The expression of different proliferation associated nuclear antigens was analyzed using a washless double-staining method and flow cytometry. It is a simple and rapid two-step procedure which can be performed on low cell numbers. A series of hematopoietic cell lines and fresh lymphoma cells were tested and the methodology was found to be applicable to a number of nuclear antigens (PCNA, Ki-67, p105, MPM-2, fibrillarin). For PCNA, the detectability was dependent on the type of antibody used. The immunofluorescence pattern observed by microscopy was altered for antigens stained by the washless technique in comparison with the pattern obtained with fixed cells. With the washless method, detailed cell cycle analysis could be obtained by dual parameter analysis of PCNA and Ki-67.

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Section: Discussionmentioning

confidence: 98%

Flow cytometric analysis of proliferation associated nuclear antigens using washless staining of unfixed cells

Landberg

Roos

1992

Cytometry

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“…For example, a cell that has just completed mitosis and cytokinesis in the proximal column is probably migrating distally (toward the shell) as it enters the GI-phase of the next cell cycle, when PCNA levels would be expected to increase (Kurki et al, 1986(Kurki et al, , 1988. Even if the trophoblast cell exits the cell cycle and differentiates, entering Go, the half-life of PCNA is sufficiently long (approximately 20 hr; Bravo and Macdonald-Bravo, 1987) that these cells would continue to be labeled for some time. This residual PCNA that is immunolabeled, but not functioning in DNA synthesis, may lead to a n overestimation of the number of cycling cells.…”

Section: Discussionmentioning

confidence: 99%

Developmental expression of Ki‐67 antigen and proliferating cell nuclear antigen in macaque placentas

Blankenship

King

1994

Developmental Dynamics

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Placental growth is largely determined by the proliferation of cytotrophoblast cells. However, the distribution of cytotrophoblast cells engaged in the cell cycle during placental development is poorly understood. Recently, antibodies have been developed that identify two proteins directly involved with DNA synthesis: Ki-67 protein and proliferating cell nuclear antigen (PCNA). Immunolocalization of Ki-67 and PCNA provides a measure of the proliferating cells in tissues. We examined, in macaque placentas, the spatiotemporal pattern of expression of these proteins during gestation. Tissues from 24 macaque placentas collected from 22-153 days of pregnancy were prepared for paraffin sections. Standard immunoperoxidase techniques were used to identify Ki-67 and PCNA. The proteins generally co-localized, although PCNA was usually represented in more cells than Ki-67. Early in gestation the cell columns contained many labeled cells. The cytotrophoblastic shell was occupied by numerous cells with PCNA positive nuclei, but few were reactive for Ki-67. By 45 days of pregnancy the immunolabeled cells in the cell columns were concentrated in the proximal regions, adjacent to the anchoring villus tips. The number of positive cells decreased by 100 days when the cell columns were diminished, leaving the anchoring villus tips buried in the shell. Labeled cells were rarely present in the shell at late pregnancy. The single layer of cytotrophoblast cells in the chorionic plate contained numerous reactive cells throughout early and mid-gestation. After approximately 100 days the cytotrophoblast layer of the chorionic plate was stratified over large areas. Soon thereafter few cells of the chorionic plate were labeled. The chorionic villi contained reactive cytotrophoblastic cells throughout gestation. Extravillous cytotrophoblast cells invading spiral arteries were sometimes labeled for PCNA but not Ki-67. We conclude that compartments of the placenta are distinguished by specific patterns of cytotrophoblast cell proliferation. Moreover, these patterns correspond to macroscopic growth parameters of the placenta. Evidence suggests that the macaque placenta slows its rate of diametrical growth at approximately 100 days of gestation. It is at about this time that the cell columns are absorbed into the trophoblastic shell and this pool ofproliferating cells is diminished. The growth in diameter of the chorionic plate matches that of the shell. In this compartment also the architecture changes at about 100 days as the cytotrophoblast layer stratifies. This stratification may result from continued proliferation of cytotrophoblast cells when the diametrical rate of growth is decreasing. Soon thereafter, proliferation decreases in this compartment also. By contrast, labeled cells were found in chorionic villi throughout gestation. Continued villous growth and branching probably accounts for most of the increases in placental thickness and weight that occurs during late gestation. 0 1994 Wiley-Liss, Inc.

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“…These antibodies recognize the carboxyl terminus of MAP kinase and the tyrosine phosphorylated residues 196 ± 209 of human p44 MAP kinase, respectively. The proliferative cell nuclear antigen (PCNA) is a nuclear protein of 36 kDa associated with the cell cycle (Bravo and MacDonald-Bravo, 1987). Levels of PCNA were taken as an index of cell proliferation.…”

Section: Immunoblotting and Map Kinase Assaymentioning

confidence: 99%

Strict regulation of c-Raf kinase levels is required for early organogenesis of the vertebrate inner ear

et al. 1999

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Regulation of organogenesis involves a dynamic balance of the mechanisms regulating cell division, di erentiation and death. Here we have investigated the pattern of expression of c-Raf kinase in the inner ear during early developmental stages and the consequences of manipulating c-Raf levels by misexpression of c-raf viral vectors in organotypic cultures of otic vesicle explants. We found that otic vesicles expressed c-Raf and its level remained constant during embryonic days 2 and 3 (E2-E3). c-Raf activity was increased in response to insulin like growth factor-I (IGF-I) and the activation by IGF-I of the c-Raf kinase pathway was a requirement to turn on cell proliferation in the otic vesicle. Overexpression of c-raf in E2.5 explants increased the proliferative response to low serum and IGF-I and blocked di erentiation induced by retinoic acid. The increase in c-Raf levels also prevented nerve growth factor (NGF)-dependent induction of programmed cell death. Consistent with these results, the expression of a dominant negative c-Raf mutant potentiated retinoic acid action and decreased the rate of cell proliferation. We conclude that a strict control of c-Raf levels is essential for the co-ordination of the biological processes that operate simultaneously during early inner ear development.

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Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites.

Cited by 875 publications

References 31 publications

Flow cytometric analysis of proliferation associated nuclear antigens using washless staining of unfixed cells

Flow cytometric analysis of proliferation associated nuclear antigens using washless staining of unfixed cells

Developmental expression of Ki‐67 antigen and proliferating cell nuclear antigen in macaque placentas

Strict regulation of c-Raf kinase levels is required for early organogenesis of the vertebrate inner ear

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