Tandem Mass Tag-Based High-Resolution LC-MS/MS identifies free D-aspartate-induced expression of proteins linked to schizophrenia and autism spectrum disorder

Errico, F.; Russo, R.; Carrillo, F.; Nuzzo, T.; di Vito, R.; Canonico, E.; Pedone, P. V.; Di Cunto, F.; Esposito, T.; Usiello, A.; Chambery, A.

doi:10.1101/2025.04.18.649487

bioRxiv · 10.1101/2025.04.18.649487

Tandem Mass Tag-Based High-Resolution LC-MS/MS identifies free D-aspartate-induced expression of proteins linked to schizophrenia and autism spectrum disorder

Errico, F.·Russo, R.·Carrillo, F.·Nuzzo, T.·di Vito, R.·Canonico, E.·Pedone, P. V.·Di Cunto, F.·Esposito, T.·Usiello, A.·Chambery, A.

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Abstract

D-aspartate is an endogenous agonist of NMDA and mGlu5 receptors, with a distinctive spatiotemporal expression profile that peaks in the prenatal and early postnatal brain. This suggests a critical role for D-aspartate metabolism in modulating neurodevelopmental processes linked to glutamatergic neurotransmission. However, the precise mechanisms through which D-aspartate exerts its effects remain unclear. To elucidate the molecular pathways orchestrated by early D-aspartate signalling, we employed a knockin mouse model characterized by constitutive D-aspartate depletion due to the prenatal expression of its degradative enzyme, D-aspartate oxidase. Using an advanced quantitative proteomic approach based on Tandem Mass Tag isobaric labelling and nano-liquid chromatography coupled with high-resolution tandem mass spectrometry, we investigated the proteomic variations induced by D-aspartate depletion during postnatal brain development comparing Ddo knockin mice with their wild-type littermates. Our findings reveal that D-aspartate modulates the neonatal expression of proteins involved in glutamatergic neurotransmission, nervous system development, and cytoskeleton organization. Moreover, proteomic analysis identified a subset of D-aspartate-regulated proteins mapping molecular pathways associated with autism spectrum disorder and schizophrenia. These findings offer new perspectives on the complex protein networks influenced by D-aspartate metabolism in the developing brain and highlight its potential impact on cerebral function in health and psychiatric disorders.

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Errico, F., Russo, R., Carrillo, F., Nuzzo, T., di Vito, R., Canonico, E., Pedone, P. V., Di Cunto, F., Esposito, T., Usiello, A., Chambery, A.. 2025-04-20. Tandem Mass Tag-Based High-Resolution LC-MS/MS identifies free D-aspartate-induced expression of proteins linked to schizophrenia and autism spectrum disorder. https://doi.org/10.1101/2025.04.18.649487

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