Epitope analysis of antibodies recognising the cell proliferation associated nuclear antigen previously defined by the antibody Ki-67 (Ki-67 protein).

Kubbutat, Michael H.G.; Key, Göran; Duchrow, M.; Schlüter, Carsten; Flad, Hans‐Dieter; Gerdes, Johannes

doi:10.1136/jcp.47.6.524

Cited by 80 publications

(62 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…4A). We also costained with anti-Ki67, because this protein marks cycling cells (44), and RAG2 is degraded in a cell-cycle-dependent fashion (45). As expected, RAG2 staining was generally low in the Ki67 + subset (∼15%) of Lat −/− nuclei but was readily detected in most Ki67 low nondividing nuclei (Fig.…”

Section: Resultsmentioning

confidence: 56%

Peripheral subnuclear positioning suppresses Tcrb recombination and segregates Tcrb alleles from RAG2

Chan¹,

Teng²,

Corbett³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Allelic exclusion requires that the two alleles at antigen-receptor loci attempt to recombine variable (V), diversity (D), and joining (J) gene segments [V(D)J recombination] asynchronously in nuclei of developing lymphocytes. It previously was shown that T-cell receptor β (Tcrb) alleles frequently and stochastically associate with the nuclear lamina and pericentromeric heterochromatin in CD4 − CD8− thymocytes. Moreover, rearranged alleles were underrepresented at these locations. Here we used 3D immunofluorescence in situ hybridization to identify recently rearranged Tcrb alleles based on the accumulation of the DNA-repair protein 53BP1. We found that Tcrb alleles recombine asynchronously in double-negative thymocytes and that V(D)J recombination is suppressed on peripheral as compared with central Tcrb alleles. Moreover, the recombination events that did take place at the nuclear periphery preferentially occurred on Tcrb alleles that were partially dissociated from the nuclear lamina. To understand better the mechanism by which V(D)J recombination is suppressed at the nuclear periphery, we evaluated the subnuclear distribution of recombination-activating gene 2 (RAG2) protein. We found that RAG2 abundance was reduced at the nuclear periphery. Moreover, RAG2 was distributed differently from RNA polymerase II and histone H3K4 trimethylation. Our data suggest that the nuclear periphery suppresses V(D)J recombination, at least in part, by segregating Tcrb alleles from RAG proteins.T-cell development | thymus A ntigen receptor variable (V), diversity (D), and joining (J) gene segments are assembled by V(D)J recombination in immature T and B lymphocytes to generate diverse repertoires of T-cell receptors (TCRs) and B-cell receptors (BCRs), respectively (1). V(D)J recombination is initiated by the recombination-activating gene (RAG) 1 and 2 proteins, which bind to and induce double-strand breaks (DSBs) at recombination signal sequences that flank V, D, and J segments. V(D)J recombination at antigen-receptor loci is regulated according to cell lineage and developmental stage (2). In addition, at some loci V (D)J recombination is regulated to enforce allelic exclusion, so that a complete antigen-receptor protein is produced by only one allele (3, 4). However, the mechanisms that establish allelic exclusion are poorly understood.Among TCR loci, only the T-cell receptor β (Tcrb) locus is allelically excluded (5). Tcrb recombination occurs in CD4 − CD8 − double-negative (DN) thymocytes and is ordered, beginning with D β -J β rearrangement, which can occur on both alleles. Allelic exclusion then is initiated by V β -to-DJ β recombination, which is thought to occur asynchronously, i.e., on one allele at a time. This asynchrony allows thymocytes time to test each allele for the creation of an ORF. TCRβ proteins are sensed by their assembly with pre-Tα and CD3 chains to create a pre-TCR signaling complex; pre-TCR signals then suppress further Tcrb recombination and promote thymocyte proliferation and differentiation to the CD...

show abstract

Section: Resultsmentioning

confidence: 56%

Peripheral subnuclear positioning suppresses Tcrb recombination and segregates Tcrb alleles from RAG2

Chan¹,

Teng²,

Corbett³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…1C). Whereas the majority of fetal liver HPCs resided in S-G 2 -M (78%), the majority of fetal liver HSCs were in G 1 (51%), with just 34% in S-G 2 -M. Moreover, whereas virtually no HPCs were found in G 0 , as much as 14% of the HSC population appeared to be in a G 0 state, as indicated by undetectable Ki67 expression (23). Thus, despite extensive expansion and continuous proliferation, fetal liver HSCs appear to reenter a transient state of relative quiescence and prolonged G 1 transit.…”

Section: Prolonged Cell Cycle Transit Of Fetal Liver Hscs During Physmentioning

confidence: 99%

Prolonged Cell Cycle Transit Is a Defining and Developmentally Conserved Hemopoietic Stem Cell Property

Nygren

Bryder

Jacobsen

2006

The Journal of Immunology

View full text Add to dashboard Cite

Adult mouse hemopoietic stem cells (HSCs) are typically quiescent and enter and progress through the cell cycle rarely in steady-state bone marrow, but their rate of proliferation can be dramatically enhanced on demand. We have studied the cell cycle kinetics of HSCs in the developing fetal liver at a stage when they expand extensively. Despite that 100% of fetal liver HSCs divide within a 48-h period, their average cell cycle transit time (10.6 h) is twice that of their downstream progenitors, translating into a prolonged G1 transit and a period of relative quiescence (G0). In agreement with their prolonged G1 transit when compared with hemopoietic progenitors, competitive transplantation experiments demonstrate that fetal HSCs are highly enriched in G1 but also functional in S-G2-M. This observation combined with experimental data demonstrating that adult HSCs forced to expand ex vivo also sustain a uniquely prolonged cell cycle and G1 transit, demonstrate at least in part why purified HSCs at any state of development or condition are highly enriched in the G0-G1 phases of the cell cycle. We propose that a uniquely prolonged cell cycle transit is a defining stem cell property, likely to be critical for their maintenance and self-renewal throughout development.

show abstract

“…The primary antibodies used in this study were anticalretinin (1/400, Swant) (Schwaller et al, 1993), anti-NURR1 (NR4A2 -Mouse Genome Informatics; 1/100, R&D Systems) (Hoerder-Suabedissen et al, 2009), anti-PAX6 (1/200, Covance) (Marquardt et al, 2001), anti-TBR2 (1/500, Chemicon), anti-TBR1 (1/500, Abcam) (Englund et al, 2005), anti-CUX1 (1/100, Santa Cruz Biotech. ), anti-TUJ1 (1/500, Sigma) (Lee et al, 1990), anti-chondroitin sulfate (1/200, Sigma) (Bicknese et al, 1994), anti-Ki67 (1/500, BD, Pharmingen), (Kubbutat et al, 1994), antineurogenin 2 (1/100, R&D Systems) (Lo et al, 2002), anti-BrdU (1/500, BD, Pharmingen) (Dolbeare et al, 1983) and anti-CTIP2 (1/500, Abcam) (Lai et al, 2008). The secondary antibodies were Alexa 488-or 555-conjugated goat anti-mouse, anti-rabbit or anti-rat IgG (Molecular Probes).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Zinc finger genes Fezf1 and Fezf2 control neuronal differentiation by repressing Hes5 expression in the forebrain

Shimizu¹,

Nakazawa²,

Kani³

et al. 2010

Development

View full text Add to dashboard Cite

SUMMARYPrecise control of neuronal differentiation is necessary for generation of a variety of neurons in the forebrain. However, little is known about transcriptional cascades, which initiate forebrain neurogenesis. Here we show that zinc finger genes Fezf1 and Fezf2, which encode transcriptional repressors, are expressed in the early neural stem (progenitor) cells and control neurogenesis in mouse dorsal telencephalon. Fezf1-and Fezf2-deficient forebrains display upregulation of Hes5 and downregulation of neurogenin 2, which is known to be negatively regulated by Hes5. We show that FEZF1 and FEZF2 bind to and directly repress the promoter activity of Hes5. In Fezf1-and Fezf2-deficient telencephalon, the differentiation of neural stem cells into early-born cortical neurons and intermediate progenitors is impaired. Loss of Hes5 suppresses neurogenesis defects in Fezf1-and Fezf2-deficient telencephalon. Our findings reveal that Fezf1 and Fezf2 control differentiation of neural stem cells by repressing Hes5 and, in turn, by derepressing neurogenin 2 in the forebrain.

show abstract

Epitope analysis of antibodies recognising the cell proliferation associated nuclear antigen previously defined by the antibody Ki-67 (Ki-67 protein).

Cited by 80 publications

References 12 publications

Peripheral subnuclear positioning suppresses Tcrb recombination and segregates Tcrb alleles from RAG2

Peripheral subnuclear positioning suppresses Tcrb recombination and segregates Tcrb alleles from RAG2

Prolonged Cell Cycle Transit Is a Defining and Developmentally Conserved Hemopoietic Stem Cell Property

Zinc finger genes Fezf1 and Fezf2 control neuronal differentiation by repressing Hes5 expression in the forebrain

Contact Info

Product

Resources

About