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A Novel In Vitro Live-imaging Assay of Astrocyte-mediated Phagocytosis Using pH Indicator-conjugated Synaptosomes
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Autophagy-Lysosomal Axis Stimulation by Beta-Hydroxybutyrate in Astrocytes.

Perla Coronado-Monroy1, Teresa Montiel1, Lizbeth García-Velázquez1

  • 1División de Neurociencias. Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México, CP, 04510, México.

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|April 20, 2026
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Beta-hydroxybutyrate (D-BHB) enhances astrocyte autophagy and lysosomal biogenesis, improving cell survival under stress. This suggests D-BHB contributes to neuroprotection by supporting glial cells, crucial for brain health.

Keywords:
AMP kinaseKetone bodiesLysosome biogenesismTORC1

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Astrocyte function is vital for neuronal support, relying on autophagy for cellular maintenance.
  • Beta-hydroxybutyrate (D-BHB), a ketone body, shows neuroprotective effects but its impact on astrocyte autophagy is unknown.

Purpose of the Study:

  • To investigate the effect of D-BHB on autophagy induction in cultured astrocytes.
  • To explore the molecular pathways involved in D-BHB-mediated autophagy in astrocytes.
  • To determine if D-BHB-induced autophagy enhances astrocyte survival under stress.

Main Methods:

  • Cultured astrocytes were exposed to D-BHB.
  • Autophagy markers (LC3-II, SQSTM1/p62), autophagosome formation, and lysosomal parameters (TFEB, LAMP1) were assessed.
  • Key signaling pathways (SIRT1, AMPK, ULK1) and astrocyte survival under oxygen-glucose deprivation (OGD) were evaluated.

Main Results:

  • D-BHB increased LC3-II levels, autophagosome formation, and TFEB activation, while decreasing SQSTM1/p62.
  • D-BHB enhanced LAMP1 abundance and lysosomal number.
  • D-BHB-induced autophagy activation was dependent on the SIRT1/AMPK/ULK1 pathway.
  • D-BHB improved astrocyte survival during OGD.

Conclusions:

  • D-BHB activates the autophagy-lysosomal pathway in astrocytes via the SIRT1/AMPK/ULK1 signaling cascade.
  • This activation enhances astrocyte resilience to ischemic conditions.
  • Astrocytes may mediate the neuroprotective effects of D-BHB, contributing to brain injury recovery.