ROLE OF ASTROCYTIC TRPA1 IN CALCIUM DYSREGULATION UNDER PATHOLOGICAL CONDITIONS

Astrocytic calcium signalling plays a critical role in regulating neuronal excitability and maintaining brain homeostasis. Dysregulation of astrocyte function is a common feature of neurodegenerative and inflammatory conditions and is often associated with astrogliosis, neuroinflammation, and increased seizure susceptibility. Transient receptor potential ankyrin 1 (TRPA1) is a non-selective cation channel expressed in astrocytes and has been proposed to regulate basal astrocytic calcium levels; however, its functional role under pathological conditions remains incompletely understood.
In this study, we employ ex vivo brain slice and cellular models to investigate the contribution of astrocytic TRPA1 to calcium signalling under pathological stress. Using Aldh1l1-Cre TRPA1 conditional knockout mice, astrocytic calcium activity is examined under normoxic (20% O₂) and hypoxic (0% O₂) conditions, as well as following exposure to disease-related protein stimuli, including tau. Calcium imaging is used to assess astrocytic responses, and functional TRPA1 expression is further evaluated by pharmacological activation with the TRPA1 agonists JT010 and AITC. In addition, Aβ treatment is planned to model neurodegenerative conditions.
To explore the potential functional consequences of astrocytic TRPA1 alteration at the network level, pentylenetetrazol-induced seizure paradigms are used to assess seizure severity and latency. Immunohistochemical analyses are performed to evaluate glial reactivity and neuronal status, including GFAP and IBA1 for astrocytes and microglia, as well as NeuN and cFos for neuronal integrity and activation.
Together, this work aims to determine whether astrocytic TRPA1 contributes to calcium dysregulation under pathological conditions and whether such changes are associated with glial reactivity, altered neuronal excitability, and seizure-related phenotypes.