A NOVEL IN VITRO HUMAN NEURONAL MODEL OF CORTICAL SPREADING DEPOLARIZATION

Migraine is the third most prevalent neurological disorder worldwide, and its pathophysiology has been extensively investigated. Among the proposed mechanisms, cortical spreading depolarization (CSD) is the most accepted phenomenon underlying migraine, particularly migraine with aura. CSD is characterized by a slowly propagating wave of near-complete neuronal and glial depolarization accompanied by profound ionic, metabolic, and vascular changes. Importantly, CSD is not unique to migraine but has also been implicated in other neurological conditions, including stroke, traumatic brain injury, and epilepsy, highlighting its broader relevance in brain pathology.
While CSD has been characterized in animal models, studies investigating this phenomenon in human neuronal systems remain limited. To address this gap, we developed a novel in vitro human neuronal model of CSD using neurons differentiated from the SH-SY5Y neuroblastoma cell line. This model enables the investigation of CSD-related cellular mechanisms in a human neuronal context.SH-SY5Y cells were differentiated into neuron-like cells, and neuronal identity was confirmed by immunofluorescence and Western blot. CSD was induced using a combination of microneedle scratching and potassium chloride (KCl) application in neurons cultured on plates prepared for this paradigm.
Following CSD induction, pannexin channel activation was associated with increased propidium iodide uptake, indicating enhanced membrane permeability.While the neurogenic inflammatory markers IL-1β and TLR4 were not detectable under control conditions, their expression was increased in CSD-induced neurons.
This study presents a novel in vitro human neuronal model of CSD and provides a platform for investigating shared CSD-related mechanisms, with potential implications for the development of therapeutic strategies.