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The Ketone Body, β-Hydroxybutyrate Stimulates the Autophagic Flux and Prevents Neuronal Death Induced by Glucose Deprivation in Cortical Cultured Neurons

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Abstract

Glucose is the major energy substrate in brain, however, during ketogenesis induced by starvation or prolonged hypoglycemia, the ketone bodies (KB), acetoacetate and β-hydroxybutyrate (BHB) can substitute for glucose. KB improve neuronal survival in diverse injury models, but the mechanisms by which KB prevent neuronal damage are still not well understood. In the present study we have investigated whether protection by the D isomer of BHB (D-BHB) against neuronal death induced by glucose deprivation (GD), is related to autophagy. Autophagy is a lysosomal-dependent degradation process activated during nutritional stress, which leads to the digestion of damaged proteins and organelles providing energy for cell survival. Results show that autophagy is activated in cortical cultured neurons during GD, as indicated by the increase in the levels of the lipidated form of the microtubule associated protein light chain 3 (LC3-II), and the number of autophagic vesicles. At early phases of glucose reintroduction (GR), the levels of p62 declined suggesting that the degradation of the autophagolysosomal content takes place at this time. In cultures exposed to GD and GR in the presence of D-BHB, the levels of LC3-II and p62 rapidly declined and remained low during GR, suggesting that the KB stimulates the autophagic flux preventing autophagosome accumulation and improving neuronal survival.

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Abbreviations

GD:

Glucose deprivation

GR:

Glucose reperfusion

LC3:

Microtubule associated protein light chain 3

LC3-II:

Lipidated form of the microtubule associated protein light chain 3

BHB:

β-Hydoxybutyrate

D-BHB:

D isomer of β-hydoxybutyrate

3-MA:

3-Methyl adenine

KB:

Ketone bodies

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Acknowledgments

This study was performed in partial fulfillment of the requirements for the Ph.D. degree in Ciencias Biomédicas of L. Camberos-Luna at the Universidad Nacional Autónoma de México. This work was supported by Programa de Apoyo a Proyectos de Investigación e Inovación Tecnológica (PAPIIT) grant IN204213 and Consejo Nacional de Ciencia y Tecnología (CONACYT) Grant CB-239607 to LM and CONACYT scholarship to L. Camberos-Luna. Authors thank Augusto César Poot-Hernández for his help in vesicle counting.

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    Authors and Affiliations

    1. División de Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México (UNAM), AP 70-253, CP 04510, Mexico, DF, Mexico

      Lucy Camberos-Luna, Cristian Gerónimo-Olvera, Teresa Montiel, Ruth Rincon-Heredia & Lourdes Massieu

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    1. Lucy Camberos-Luna
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    2. Cristian Gerónimo-Olvera
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    Correspondence to Lourdes Massieu.

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    Special Issue: In Honor of Philip Beart.

    Lucy Camberos-Luna and Cristian Gerónimo-Olvera have equally contributed to this work.

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    Camberos-Luna, L., Gerónimo-Olvera, C., Montiel, T. et al. The Ketone Body, β-Hydroxybutyrate Stimulates the Autophagic Flux and Prevents Neuronal Death Induced by Glucose Deprivation in Cortical Cultured Neurons. Neurochem Res 41, 600–609 (2016). https://doi.org/10.1007/s11064-015-1700-4

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