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Methods to study primary cilia and autophagy in the brain.

Laura de Las Heras-García1, Irati Zabalegui2, Olatz Pampliega1

  • 1Departamento de Neurociencias, Universidad del País Vasco (UPV/EHU), Leioa, Spain; Achucarro Basque Center for Neurosciences, Leioa, Spain.

Methods in Cell Biology
|May 10, 2023
PubMed
Summary

Autophagy and primary cilia (PC) are vital for brain cell homeostasis. This chapter details methods for studying their interplay in brain slices, aiding research into neurological conditions.

Keywords:
AstrocyteAutophagyImmunohistochemistryNeuronPrimary cilium

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Area of Science:

  • Cell Biology
  • Neuroscience
  • Molecular Biology

Background:

  • Autophagy is a critical cellular recycling process essential for maintaining homeostasis, particularly in post-mitotic cells like neurons.
  • The primary cilium (PC) functions as a cellular sensory organelle, and its role alongside autophagy is increasingly recognized in various cell types, including brain cells.
  • Dysregulation of autophagy and PC is implicated in neurological disorders.

Purpose of the Study:

  • To review the current understanding of the interplay between autophagy and primary cilia (PC) in the brain.
  • To highlight the implications of this interplay in both healthy and diseased brain states.
  • To provide practical immunofluorescence methods for studying autophagy and PC in brain tissue.

Main Methods:

  • Immunofluorescence techniques are described for visualizing autophagy and PC.
  • Methods cover tissue preparation, confocal microscopy, and image analysis.
  • Specific protocols for assessing changes in PC length/number and autophagy markers are detailed.

Main Results:

  • The chapter outlines how to practically assess autophagy and PC status in brain slices.
  • It demonstrates how to correlate changes in these pathways with cellular health and disease.
  • User-friendly protocols are provided for reproducible analysis.

Conclusions:

  • Studying the interplay between autophagy and PC is crucial for understanding brain function and disease.
  • The provided immunofluorescence methods offer a practical approach for researchers.
  • This work facilitates further investigation into therapeutic strategies targeting these pathways in neurological conditions.