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Neuronal Subtype-Specific Expression of γ-Enolase: Its Role in Neuronal Differentiation.

Selena Horvat1, Urša Pečar Fonović1, Nace Zidar2

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Gamma-enolase supports neuronal differentiation, especially in cholinergic-like neurons. Its activity is regulated by cathepsin X, offering potential for neuroregeneration therapies.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Neuronal differentiation is vital for nervous system development, influenced by neurotrophic factors.
  • Gamma-enolase (ENO2), a neuron-specific enzyme, has shown neurotrophic-like properties.
  • The specific role of gamma-enolase in different neuronal subtypes is not well understood.

Purpose of the Study:

  • To investigate the role of gamma-enolase in the differentiation of dopaminergic-, cholinergic-, and adrenergic-like neuronal cells.
  • To determine how gamma-enolase influences neuronal morphological maturation and cytoskeletal markers.
  • To explore the regulatory interaction between gamma-enolase and cathepsin X in neuronal differentiation.

Main Methods:

  • Quantitative analysis of gamma-enolase expression in differentiated neuronal cells.
  • Assessment of neurite outgrowth and beta-tubulin expression following gamma-enolase manipulation (full-length, truncated, peptide, silencing).
  • Confocal microscopy to visualize co-localization of gamma-enolase and cathepsin X; inhibition of cathepsin X.

Main Results:

  • Gamma-enolase expression significantly increased upon neuronal differentiation, with highest levels in cholinergic-like neurons.
  • Full-length gamma-enolase and its C-terminal peptide enhanced neurite outgrowth and beta-tubulin expression.
  • Silencing gamma-enolase reduced neurite length, confirming its role in maturation.
  • Cathepsin X cleaves gamma-enolase, reducing its neurotrophic effects; inhibition of cathepsin X preserved active gamma-enolase and promoted differentiation.

Conclusions:

  • Gamma-enolase plays a significant role in neuronal differentiation, particularly in cholinergic-like neurons.
  • Cathepsin X acts as a negative regulator of gamma-enolase activity.
  • The gamma-enolase/cathepsin X interaction presents potential therapeutic targets for neuroregeneration.