Nuclear size regulation: from single cells to development and disease

Edens, Lisa J.; White, Karen H.; Jevtić, Predrag; Li, Xiaoyang; Levy, Daniel L.

doi:10.1016/j.tcb.2012.11.004

Cited by 111 publications

(89 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…These limits corresponded to approximately 20, 40, and 50 nuclei per colony, respectively, assuming a mean nuclear diameter of 10 μm. 22,23 As shown in Figure 3C, the area covered by different colony sizes followed the same trend and a similar Nut-3a IC 50 . Indeed, we obtained IC 50 values of 1.6±0.14 μM, 1.5±0.05 μM, and 1.4±0.10 μM for colonies with diameters of >200 μm, >400 μm, and >500 μm, respectively.…”

Section: Drug Dose-response Curve Validationsupporting

confidence: 61%

Open Access to High-Content Clonogenic Analysis

2015

View full text Add to dashboard Cite

Image-processing programs are used to identify and classify eukaryotic cell colonies as spots following seeding at low density on dishes or in multiwell plates. The output from such approaches, however, is generally limited to 1-2 parameters, and there is no ability to extract phenotypic information at the single colony level. Furthermore, there is a lack of userfriendly pipelines for analysis of clonogenicity in the context of high-content analysis. This article describes an experimental and multiparametric image analysis workflow for clonogenic assays in multiwell format, named the Colony Assay Toolbox (CAT). CAT incorporates a cellular-level resolution of individual colonies and facilitates the extraction of phenotypic information, including the number and size of colonies and nuclei, as well as morphological parameters associated with each structure. Furthermore, the pipeline is capable of discriminating between colonies composed of senescent and nonsenescent cells. We demonstrate the accuracy and flexibility of CAT by interrogating the effects of 2 preclinical compounds, Nutlin3a and ABT-737, on the growth of human osteosarcoma cells. CAT is accessible to virtually all laboratories because it uses common wide-field fluorescent microscopes, the open-source CellProfiler program for colony image analysis, and a single fluorescent dye for all the segmentation steps.

show abstract

Section: Drug Dose-response Curve Validationsupporting

confidence: 61%

Open Access to High-Content Clonogenic Analysis

2015

View full text Add to dashboard Cite

show abstract

“…Because the correlation between genome and cell size also includes a correlation with the size of the nucleus, older hypotheses for this multitrait correlation also included an active role for nucleus size in the proposed mechanisms, linking all three traits. However, recent data suggests that the size of the nucleus responds directly to cytoplasmic volume, irrespective of the amount of DNA (Edens et al 2013); a direct connection between genome and cell size is consistent with this model of nuclear size determination.…”

Section: Genome Biology and Cell Sizesupporting

confidence: 59%

Genome Biology and the Evolution of Cell-Size Diversity

Mueller

2015

Cold Spring Harb Perspect Biol

View full text Add to dashboard Cite

Cell size is highly variable among different species across the Tree of Life. For decades, biologists have generated hypotheses to explain this variation, in many cases, drawing on the correlations that exist among cell size, genome size, nucleus size, and various physiological and developmental parameters. In recent years, our understanding of the molecular processes that generate variation in genome size over evolutionary time, as well as the processes that maintain homeostasis in cell size over ontogenetic time, has increased dramatically. The goal of this article is to highlight how information from these fields can be integrated to generate new hypotheses to explain cell-size diversity.

show abstract

“…To determine how nuclear size and the N/C volume ratio might impact developmental progression, we sought multiple approaches employing different mechanisms to manipulate nuclear size in the embryo, utilizing factors known to regulate nuclear size: importins, lamins, and reticulons [21, 22]. Previous work in Xenopus egg extracts and early embryos demonstrated that rates of nuclear import influence nuclear size, with the levels of importin α being particularly important [23].…”

Section: Resultsmentioning

confidence: 99%

Nuclear Size Scaling during Xenopus Early Development Contributes to Midblastula Transition Timing

2015

View full text Add to dashboard Cite

Summary Early Xenopus laevis embryogenesis is a robust system for investigating mechanisms of developmental timing. After a series of rapid cell divisions with concomitant reductions in cell size, the first major developmental transition is the midblastula transition (MBT), when zygotic transcription begins and cell cycles elongate [1-3]. While the maintenance of a constant nuclear-to-cytoplasmic (N/C) volume ratio is a conserved cellular property [4-7], it has long been recognized that the N/C volume ratio changes dramatically during early Xenopus development [8]. We investigated how changes in nuclear size and the N/C volume ratio during early development contribute to the regulation of MBT timing. While previous studies suggested a role for the N/C volume ratio in MBT timing [1, 9-13], none directly tested the effects of altering nuclear size. In this study, we first quantify blastomere and nuclear sizes in X. laevis embryos, demonstrating that the N/C volume ratio increases prior to the MBT. We then manipulate nuclear volume in embryos by microinjecting different nuclear scaling factors, including import proteins, lamins, and reticulons. Using this approach, we show that increasing the N/C volume ratio in pre-MBT embryos leads to premature activation of zygotic gene transcription and early onset of longer cell cycles. Conversely, decreasing the N/C volume ratio delays zygotic transcription and leads to additional rapid cell divisions. While the DNA-to-cytoplasmic ratio has been implicated in MBT timing [1, 9-18], our data show that nuclear size also contributes to the regulation of MBT timing, demonstrating the functional significance of nuclear size during development.

show abstract

Nuclear size regulation: from single cells to development and disease

Cited by 111 publications

References 65 publications

Open Access to High-Content Clonogenic Analysis

Open Access to High-Content Clonogenic Analysis

Genome Biology and the Evolution of Cell-Size Diversity

Nuclear Size Scaling during Xenopus Early Development Contributes to Midblastula Transition Timing

Contact Info

Product

Resources

About