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Fig. S3

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ZDB-IMAGE-170630-9
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Figures for Boulanger-Weill et al., 2017
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Fig. S3

Rrelated to Figure 5. Spontaneous activity correlations between newborn-labeled and mature tectal neurons

(A) Left, example of an optical plane of the optic tectum containing a newborn-labeled neuron (red) chronically imaged from 2 to 4 dpe. Neurons are color-coded according to their pair-wise correlation values with the newborn-labeled neuron. Color scale bar: Pair-wise correlations between the newborn-labeled neuron and the rest of the tectal neurons. Scale bars: 50 μm. Insets: Morphology of the newborn-labeled neuron. Scale bars: 20 μm. Right, the percentage of significant correlations between the developing newborn-labeled neurons and the rest of the tectal cells, with respect to the physical distance between the neurons, for 2 (red), 3 (black) and 4 (blue) dpe. Neurons involving few pixels (less than 5) or with unstable baselines (rapid changes generally indicating movement artifacts) were left unlabeled.

(B) The developmental dynamics of the relationship between the mean of significant correlations and the physical distance between the neurons, for the newborn-labeled neurons (red) and the mature neurons (blue), from 1 to 4 dpe. Confidence intervals: SEM. Gray shaded areas: significant differences between newborn and mature curves (p<0.05, Mann-Whitney U tests).

(C) Examples of 6 spontaneous neuronal assemblies showing their topographies. Magenta: neurons within an assembly.

(D) The maximal principal component loadings for newborn-labeled and mature neurons. Maximal principal component loadings reflect the level of prediction of the activity of a neuron by the activity of the assembly which contains it, assuming that each neuron belongs only to one assembly. *: p<0.05. ***: p<0.001. Error bars: S.E.M, newborn neurons: n= 6, 15, 21, 15; mature neurons: n=1962, 3564, 5296, 3969, from 6, 10, 17 and 11 larvae, respectively.

(E-G) Top, example of an optical section of the optic tectum showing neurons color-coded according to X2 distance (high values indicate low level of significant correlations), the frequency of the spontaneous Ca2+ events (middle), and the amplitude of the spontaneous Ca2+ events (right). The X2 distance is the euclidean distance between the average distribution represented in Figure 5C (mature neurons, blue curve) and the distribution of each neuron, for distances shorter than 60μm. High X2 values indicate neurons that are isolated from others (do not show significant correlations with other neurons). Bottom, averaged distributions of the measured parameters, across the normalized coordinates along the caudo-rostral axis, for all neurons, from 5 to 8 dpf (n=17678 neurons, from 44 larvae). Confidence interval: SEM. Changes in the X2 distance across the caudorostral axis were significant, while the frequency and amplitude of the spontaneous Ca2+ events did not show significant changes (p=4.12x10-6, p=0.55 and p=0.26, respectively, Kruskal-Wallis tests). Neurons involving few pixels (< 5) or with unstable baselines (rapid changes generally indicating movement artifacts) were left unlabeled.

Acknowledgments
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