Abstract
Circulation of cerebrospinal fluid (CSF) plays an important role during development. In zebrafish embryo, the flow of CSF has been found to be bidirectional in the central canal of the spinal cord. In order to compare conditions and genetic mutants across each other, we recently automated the quantification of the velocity profile of exogenous fluorescent particles in the CSF. We demonstrated that the beating of motile and tilted cilia localized on the ventral side of the central canal was sufficient to generate locally such bidirectionality. Our approach can easily be extended to characterize CSF flow in various genetic mutants. We provide here a detailed protocol and a user interface program to quantify CSF dynamics. In order to interpret potential changes in CSF flow profiles, we provide additional tools to measure the central canal diameter, characterize cilia dynamics and compare experimental data with our theoretical model in order to estimate the impact of cilia in generating a volume force in the central canal. Our approach can also be of use for measuring particle velocity in vivo and modeling flow in diverse biological solutions.
Highlights
We demonstrated that the beating of motile and tilted cilia localized on the ventral side of the central canal was sufficient to generate locally such bidirectionality
In order to interpret potential changes in cerebrospinal fluid (CSF) flow profiles, we provide additional tools to measure the central canal diameter, characterize cilia dynamics and compare experimental data with our theoretical model in order to estimate the impact of cilia in generating a volume force in the central canal
[Background] In zebrafish embryos, cerebrospinal fluid (CSF) flow is observable from 24 h post fertilization in the central canal of the spinal cord (Sternberg et al, 2018) and later extends to the brain ventricles (Olstad et al, 2019)
Summary
To achieve CSF flow profiles in the central canal of the spinal cord, we inject in brain ventricles a solution of 20 nm fluorescent beads diluted in artificial CSF (aCSF). Acquire a single image of TexasRed injected in the brain ventricles using a green laser for excitation to reliably measure the central canal diameter.
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