Dynamics and heterogeneity of a fate determinant during transition towards cell differentiation
- Northwestern University, United States
- Howard Hughes Medical Institute, University Northwestern, United States
- University of Chicago, United States
Figures
Figure 1 with 4 supplements
Development and patterning of the compound eye.
(A) Differentiation is initiated in the developing eye by the MF, which moves across the eye epithelium. On the furrow’s posterior side, G1-arrested progenitor cells undergo differentiation (light …
Figure 1—figure supplement 1
Expression and activity of the Yan-YFP transgene.
Anterior is to left. (A) Adult eye of animal carrying the Yan-YFP transgene in a yan null mutant background (yanER443/yanE884). (B) Anti-Yan fluorescence pattern (left) compared to Yan-YFP …
Figure 1—figure supplement 2
Segmentation and identification of cell nuclei in eye discs.
(A) An optical section of His2Av-mRFP fluorescence after nuclear segmentation. Segmented contours are shown in yellow, and the inset shows a magnified view of the region bounded by white rectangle. …
Figure 1—figure supplement 3
Accuracy of cell-type identification.
Cells were identified by nuclear position and shape in the His2Av-mRFP channel with the marker protein channel turned off. Identifications were then compared to the cell-type specific markers, as …
Figure 1—figure supplement 4
His2Av-mRFP fluorescence properties.
(A,B) Heat maps of fluorescence intensities in the red (A) and yellow (B) channels for an optical section of a disc expressing only His2Av-mRFP. Intensities are averaged in bins of 32 x 32 pixels …
Figure 2 with 1 supplement
Dynamics of Yan-YFP in eye cells.
(A) Average time at which initiation of differentiation is first detected by apical migration of committing cell nuclei. Time zero is set to when R8 differentiation initiates. Differentiation …
Figure 2—figure supplement 1
Mapping identified nuclei within XY space of eye discs.
Anterior is to left for each panel. (A–H) Maps of identified nuclei in an eye disc. Progenitor cell nuclei (A); R8 cell nuclei (B–H); R2 and R5 cell nuclei (C); R3 and R4 cell nuclei (D); R1 and R6 …
Figure 3 with 1 supplement
EGFR/Ras and Pnt regulate Yan-YFP levels.
(A–D) Moving averages of Yan-YFP in different cell types. Shown with shading is the standard error of the mean for each moving average. (A,C) Wildtype and EGFRts mutants incubated at the …
Figure 3—figure supplement 1
Yan-YFP levels in R2/R5 correlate with distance from R8 cells when EGFR is active.
Correlation between log-transformed detrended Yan-YFP levels in R2 and R5 nuclei and their distances from the nearest R8 nucleus as a function of developmental time. (A,B) The Pearson correlation …
Figure 4 with 2 supplements
Cell state transitions are unaffected by Yan-YFP gene copy number.
(A–C) Moving averages of Yan-YFP in eye discs containing two versus four copies of the Yan-YFP transgene. (A) R2/R5 and progenitor cells. (B) R3/R4 and progenitor cells. (C) R7 and progenitor cells. …
Figure 4—figure supplement 1
Model fitting Yan-YFP decay in eye disc cells.
(A) Progenitors, (B) R8 cells, (C) R2 and R5 cells, (D) R3 and R4 cells, (E) R1 and R6 cells, (F) R7 cells. For each dataset, four models are shown: linear, exponential, second-order polynomial, and …
Figure 4—figure supplement 2
Exponential decay models.
Yan-YFP levels in cells within a representative eye disc. Each cell type is color-coded as indicated. (A) Progenitors with fitted Hill (blue line) and exponential decay functions (black line) …
Figure 5 with 2 supplements
Yan protein half-life is largely unaffected by yan gene copy number.
Exponential decay of Yan-YFP levels. Note the robustness of Yan-YFP half-lives across replicates and yan gene copy number. Note also how half-lives are nearly twice as long for progenitor cells …
Figure 5—figure supplement 1
Mean half-lives for Yan-YFP decay versus yan gene copy number (2, 4, 6x).
Error bars represent 95% confidence intervals for estimation of the mean. Each estimation of the mean is based on bootstrapping data from one disc sample. Four discs were analyzed for each copy …
Figure 5—figure supplement 2
Compound eyes of adults carrying two, four, or six copies of the yan gene.
Anterior is to left.
Figure 6 with 4 supplements
Noise in Yan-YFP expression is highly dynamic.
Moving averages of Yan-YFP levels and noise (detrended fluctuations) for (A) progenitors, (B) R2/R5, (C) R3/R4, (D) R1/R6, and (E) R7 cells. (F) Comparative noise dynamics for all cells analyzed in …
Figure 6—figure supplement 1
Comparison of methods to measure Yan-YFP noise based on sensitivity to window size.
(A–H) Yan-YFP levels are analyzed in sliding windows of size 20, 150, 250, or 500 progenitor cells. Noise is estimated by detrended fluctuation (A,C,E,G) or coefficient of variation (B,D,F,H) within …
Figure 6—figure supplement 2
Measurement of Yan-YFP noise in all cell types.
(A,C,E,G,I,K) Yan-YFP noise is estimated by detrended fluctuations for progenitors (A), R8 (C), R2/R5 (E), R3/R4 (G), R1/R6 (I), and R7 cells (K). (B,D,F,H,J,L) Yan-YFP noise is estimated by the …
Figure 6—figure supplement 3
Comparison of Yan-YFP and endogenous Yan protein dynamics.
(A,C,E) His2Av-mRFP discs stained with anti-Yan antibody to measure endogenous Yan. (B,D,F) Yan-YFP measurements of His2Av-mRFP Yan-YFP discs in which endogenous Yan has been deleted. (A,B) Moving …
Figure 6—figure supplement 4
Moving averages of anti-Pros fluorescence levels and noise (coefficient of variation) in R7 cells.
https://doi.org/10.7554/eLife.08924.022
Figure 7
Summary of analysis.
(A) Top: a hypothetical bistable behavior would be where Yan is in a stable high state within progenitor cells and in a stable low state within differentiated cells. Bottom: the observed behavior of …
