Journal: Journal of Oncology

Article Title: Genotypic and Phenotypic Variables Affect Meiotic Cell Cycle Progression, Tumor Ploidy, and Cancer-Associated Mortality in a brca2 -Mutant Zebrafish Model

doi: 10.1155/2019/9218251

Figure Lengend Snippet: brca2 and tp53 mutations alter distribution of cells according to DNA content in adult zebrafish testes. (a) Comparison of cell progression through meiosis and mitosis with corresponding DNA content (designated as 1C, 2C, or 4C). BRCA2 participates in DNA repair during prophase I of meiosis I and performs multiple functions between late G2 and M phases of mitosis, as indicated by the positions of the yellow ovals. ((b)–(e)) Propidium iodide (PI) fluorescence histograms of testes derived from wild type (b), tp53 m/m (c), brca2 m/m (d), and brca2 m/m;tp53 m/m zebrafish testes (e). (f) Mean percent of gated cells clustered by DNA content for wild type, tp53 m/m , brca2 m/m , and brca2 m/m;tp53 m/m zebrafish testes. The mean percent of gated cells for each DNA content category is indicated. (g) Comparison of testicular morphology in wild type, tp53 m/m , brca2 m/m , and brca2 m/m;tp53 m/m zebrafish, Toluidine blue stain. Insets show type A (A, red) and type B (B, yellow) spermatogonia. Orange arrows indicate representative regions of stromal tissue and orange asterisks indicate examples of blood vessels within the stroma. (h) Comparison of the mean number of spermatogonia per 400X field (see Materials and Methods) in wild type, tp53 m/m , brca2 m/m , and brca2 m/m;tp53 m/m zebrafish testes. The mean numbers of type A and type B spermatogonia are shown in the appropriate portion of each column. SG, spermatogonia; ∗ , p = 0.01-0.05; ∗∗ , p = 0.001–0.01; ∗∗∗ , p = 0.0001–0.01; ∗∗∗∗ , p < 0.0001. Error bars represent the range of the data. See for specific p-values.

Article Snippet: After fixation, cell suspensions were washed with 1X PBS and stained with Propidium Iodide staining solution containing RNase (Cellometer PI Cell Cycle Kit, CSK-0112, Nexcelom, Lawrence, MA).

Techniques: Fluorescence, Derivative Assay, Staining

Journal: Journal of Oncology

Article Title: Genotypic and Phenotypic Variables Affect Meiotic Cell Cycle Progression, Tumor Ploidy, and Cancer-Associated Mortality in a brca2 -Mutant Zebrafish Model

doi: 10.1155/2019/9218251

Figure Lengend Snippet: brca2 mutation does not alter DNA content of nonneoplastic somatic zebrafish tissue. (a) Comparison of G0/G1 peak propidium iodide (PI) fluorescence intensity values for nonneoplastic somatic cells (upper panel) and cancer cells (lower panel) derived from 49 brca2 m/m;tp53 m/m (black circles) and 50 tp53 m/m (white circles) cancer-bearing zebrafish. Each circle indicates the G0/G1 peak value for a single sample. Two matched samples (nonneoplastic somatic cells and cancer cells) were analyzed from each individual zebrafish. For every individual zebrafish, the matched nonneoplastic somatic cell sample and cancer cell sample were analyzed in the same experiment. The experiment number indicated on the x -axis refers to each independent cell cycle analysis. In experiment 23, the cancer cell sample was excluded (see Materials and Methods), and the G0/G1 peak is only reported for the matched nonneoplastic somatic cell sample. (b) Normal distribution of G0/G1 peak PI fluorescence intensity values for nonneoplastic somatic cells derived from brca2 m/m;tp53 m/m and tp53 m/m zebrafish. In the normal quantile plot, filled black circles represent individual data points and dashed red lines indicate the Lilliefores confidence bounds. In the outlier box plot, the vertical line represents the median sample value; the diamond contains the mean and upper and lower 95% of the mean; the box ends represent the 25 th and 75 th quantiles; the whiskers extend to the outermost data points; and the red bracket indicates the shortest half (most dense 50% of observations). In the histogram, vertical bars represent G0/G1 peak intensity values by bin and the overlying red curve fits a smooth curve using nonparametric density estimation.

Article Snippet: After fixation, cell suspensions were washed with 1X PBS and stained with Propidium Iodide staining solution containing RNase (Cellometer PI Cell Cycle Kit, CSK-0112, Nexcelom, Lawrence, MA).

Techniques: Mutagenesis, Fluorescence, Derivative Assay, Cell Cycle Assay

Journal: Journal of Oncology

Article Title: Genotypic and Phenotypic Variables Affect Meiotic Cell Cycle Progression, Tumor Ploidy, and Cancer-Associated Mortality in a brca2 -Mutant Zebrafish Model

doi: 10.1155/2019/9218251

Figure Lengend Snippet: brca2 mutation status and sex alter ploidy outcomes in zebrafish cancers. (a) Representative propidium iodide (PI) fluorescence histograms from cancers that are diploid or aneuploid or exhibit complex aneuploidy. Software-identified diploid populations are depicted in red; aneuploid populations are depicted in blue and green. ((b)–(d)) Ploidy outcomes segregated by brca2 genotype alone (b) and in combination with tumor location (c) or sex (d). ((e)–(g)) Kaplan-Meier survival curves for the total study population (e), the brca2 m/m;tp53 m/m cohort (f), and the tp53 m/m cohort (g). (h) Distribution of ages at tumor diagnosis segregated by brca2 genotype and tumor ploidy. Median ages at tumor diagnosis are indicated by a red bar and are shown in red text. (i) Distribution of ages at tumor diagnosis segregated by brca2 genotype, sex, and tumor ploidy. Median ages at tumor diagnosis are indicated by a red bar and shown in red text.

Article Snippet: After fixation, cell suspensions were washed with 1X PBS and stained with Propidium Iodide staining solution containing RNase (Cellometer PI Cell Cycle Kit, CSK-0112, Nexcelom, Lawrence, MA).

Techniques: Mutagenesis, Fluorescence, Software