Inhibition of calcium-sensing receptor by its antagonist protects dopaminergic neurons from MPTP/MPP+-induced neurotoxicity via regulating mitochondrial function, autophagy, and apoptosis in vivo and in vitro

Parkinson's disease (PD)‌ is characterized by progressive degeneration of dopaminergic neurons. The role of calcium-sensing receptor (CaSR) in the pathogenesis of PD remains poorly understood. We employed the CaSR Antagonist NPS-2143 in bothin vivoandin vitroexperiments to investigate the therapeutic potential of CaSR modulation. Our findings revealed that MPTP/MPP⁺ exposure significantly upregulated CaSR expression. Functionally, CaSR overexpression exacerbated intracellular CA²⁺ dyshomeostasis under MPTP/MPP⁺ toxicity. Inhibition of CaSR with NPS-2143 demonstrated marked neuroprotection, evidenced by improved motor function and preservation of dopaminergic neurons in MPTP-treated mice, alongside reduced cellular Apoptosis in MPP⁺-injured MN9D cells. Mechanistically, NPS-2143 enhanced Autophagy in MPP⁺-exposed cells while reversing 3-MA-induced Autophagy suppression. Furthermore, NPS-2143 mitigated mitochondrial dysfunction, as shown by reduced Reactive Oxygen Species accumulation, restored mitochondrial membrane potential, and normalized ATP production in MPP⁺-treated cells. These results collectively demonstrate that CaSR antagonism protects dopaminergic neurons through coordinated regulation of mitochondrial homeostasis, autophagic flux, and apoptotic pathways. Our study highlights CaSR as a promising therapeutic target for PD prevention and identifies NPS-2143 as a potential neuroprotective agent targeting CaSR.