20.500.12556/RUL-181041 Neuronal subtype-specific expression of γ-enolase its role in neuronal differentiation Neuronal differentiation into specific subtypes is crucial for nervous system development and function, guided by neurotrophic factors. γ-Enolase, a neuron-specific glycolytic enzyme, exhibits neurotrophic-like properties and supports neuronal differentiation; however, its role in specific neuronal subtypes remains unknown. Here, we investigate the role of γ-enolase in differentiation dopaminergic-, cholinergic-, and adrenergic-like neuronal cells. Our results demonstrate that γ-enolase expression is significantly upregulated in differentiated cells, with the highest expression observed in cholinergic-like neurons. Full-length γ-enolase, compared to its truncated form, promoted enhanced neurite outgrowth and increased β-tubulin, a cytoskeletal marker. Conversely, silencing endogenous γ-enolase significantly reduced neurite length, confirming its essential role in driving neuronal morphological maturation. Furthermore, a γ-enolase-derived peptide corresponding to the active C-terminus of γ-enolase significantly promoted neurite outgrowth and increased β-tubulin expression, particularly in cholinergic-like neuronal cells. Notably, γ-enolase activity is regulated by cathepsin X, a lysosomal peptidase that cleaves γ-enolase at its C-terminus, reducing its neurotrophic effects. Confocal microscopy revealed increased co-localization of γ-enolase and cathepsin X in differentiated neuronal cells, emphasizing their interaction in cholinergic-like neurons. Inhibiting cathepsin X preserved active γ-enolase, promoted neuronal differentiation, and altered cytoskeletal marker expression. These findings suggest an important role for γ-enolase in cholinergic-like neuronal cells and propose cathepsin X as a regulatory modulator of γ-enolase activity, suggesting novel therapeutic strategies for neuroregeneration. cathepsin x regulation enolase isoforms neuronal specific subtype neurotrophic activity γ-enolase regulacija katepsina X ozooblike enolaze nevronsko specifični podtip nevrotrofična aktivnost γ-enolaza katepsin X true false true Angleški jezik Slovenski jezik Članek v reviji 2026-03-23 14:59:21 2026-03-23 14:59:27 2026-03-24 04:26:38 0000-00-00 00:00:00 2026 0 0 22 str. iss. 1, art. 4 Vol. 28 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.152.34 1559-1174 10.1007/s12017-025-08902-9 272309507 s12017-025-08902-9_(2).pdf s12017-025-08902-9_(2).pdf 1 12B008136EBCFF9A4A4D8E8AD5285212 33201e0a8c92b87dbdced4cf79fc030b1186eb19db141f7f8b5b7bf56d68d4de ad2cd926-26c1-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=231137 https://link.springer.com/article/10.1007/s12017-025-08902-9 1 66ab2212-26c0-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=231135 Fakulteta za farmacijo 0 0 0