Spatiotemporal Mapping of Brain Cilia Reveals Region-Specific Oscillation of Length and Orientation
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Abstract
In this study, we conducted high-throughput spatiotemporal analysis of primary cilia length and orientation across 22 mouse brain regions. We developed automated image analysis algorithms, which enabled us to examine over 10 million individual cilia, generating the largest spatiotemporal atlas of cilia. We found that cilia length and orientation display substantial variations across different brain regions and exhibit fluctuations over a 24-hour period, with region-specific peaks during light-dark phases. Our analysis revealed unique orientation patterns of cilia at 45° intervals, suggesting that cilia orientation within the brain is not random but follows specific patterns. Using BioCycle, we identified circadian rhythms of cilia length in five brain regions: nucleus accumbens core, somatosensory cortex, and three hypothalamic nuclei. Our findings present novel insights into the complex relationship between cilia dynamics, circadian rhythms, and brain function, highlighting cilia crucial role in the brain’s response to environmental changes and regulation of time-dependent physiological processes.
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. The cilia were imaged using the Keyence BZ-9000 fluorescence microscopewith a 20x objective lens
It's great you have a way to measure cilia in a straightforward way! Could you include here the resolution of your microscope and how that compares to the size of cilia?
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Cartesian axes (0, 45, 90, 135, 180, 225, 270, and 315), but not values inbetween (Fig. 3a,b
This is a remarkable observation! Is it possible there is some aspect of the imaging, registration, or data analysis that is leading to the strong alignment along cartesian axes? Perhaps in the way the bounding rectangle is drawn?
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averaged at 5.4 ± 0.01 μm
Based on the long right tail for the cilia length distributions, it might be more helpful to report and compare median lengths for the whole brain and each brain region.
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153,448 (in RS) to 977,129 (in DMH)
The wide range in cilia you see between these regions is interesting! Could you also report these numbers normalized to the amount of data you included so it is easier to interpret? For instance to the area of the brain region analyzed, the number of samples included per region, the number of cells in the brain region, etc.
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a specialized program to assess the length and angleof cilia in brain sections from an extensive sample size of over 50 mice (at least four mice per time point)
This is an impressive amount of data! It makes sense that you would need an automated way to quantify cilia from this many mice and brain regions. It would helpful to assess how this program works, is it possible it includes specific types of noise or misses some types of cilia? Could you take a subset of your data and compare the output of your automatic software to a manual analysis?
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