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The primary cilium, made up of microtubules, acts as antennae on the cell surfaces for relaying external stimuli into the cells. These fine hair-like structures are present, generally one per cell. These are non-motile cilia in a 9+0 microtubules arrangement, where the central pair of microtubules are absent. The primary cilia arise from the basal body embedded in the cell membrane. Intraflagellar transport (IFT) carries requisite proteins from the cytoplasm to the cilium because the primary...
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Using Primary Neurosphere Cultures to Study Primary Cilia
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Using Primary Neurosphere Cultures to Study Primary Cilia.

Issei S Shimada1, Hemant Badgandi2, Bandarigoda N Somatilaka2

  • 1Department of Cell Biology, University of Texas Southwestern Medical Center; Issei.Shimada@utsouthwestern.edu.

Journal of Visualized Experiments : Jove
|April 28, 2017
PubMed
Summary

Primary cilia are crucial for neural stem cell proliferation and neuron differentiation. This study presents neurosphere culturing methods to study primary cilia in neural stem/progenitor cells and neurons.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Developmental Biology

Background:

  • Primary cilia are essential for neural stem/progenitor cell proliferation and neuronal differentiation.
  • Differentiated neurons utilize primary cilia for signaling pathways, including Sonic Hedgehog and G-protein-coupled receptors.
  • Primary cilia defects are linked to brain developmental anomalies and cognitive deficits.

Purpose of the Study:

  • To establish and validate neurosphere-based culturing methods for studying primary cilia in neural stem/progenitor cells and neurons.
  • To investigate ciliogenesis and ciliary trafficking in neural contexts.
  • To enable the study of cilia-regulated signaling pathways in neural development and function.

Main Methods:

  • Utilized primary neurosphere cultures for mixed neural stem/progenitor cells.
  • Demonstrated amenability to multiple passages and freeze-thaw cycles.
  • Confirmed the presence of primary cilia and relevant signaling molecules in cultured neural stem/progenitor cells and differentiated neurons.

Main Results:

  • Neurosphere-derived neural stem/progenitor cells and differentiated neurons exhibit primary cilia in culture.
  • Key signaling molecules for ciliary function are localized within these primary cilia.
  • Established methods for studying ciliogenesis and ciliary trafficking in these neural cell types.

Conclusions:

  • Neurosphere-based culturing is a robust method for studying primary cilia in neural stem/progenitor cells and neurons.
  • These methods facilitate research into cilia-regulated pathways like Sonic Hedgehog and GPCR signaling in neural contexts.
  • This approach aids in understanding the role of primary cilia in neural development and function, potentially informing treatments for related disorders.