PDZD8 Dysregulation Mediates RVLM Neuronal Hyperexcitation Via Activation of Ca2+-Calpain-2 Signaling in Stress-Induced Hypertension

Neuronal hyperexcitation in the rostral ventrolateral medulla (RVLM) is crucial in the pathogenesis of stress-induced hypertension (SIH). PDZD8 connects endoplasmic reticulum (ER) to mitochondria, and is involved in SIH through regulating RVLM neuronal mitochondrial physiological function. However, the underlying mechanisms of the PDZD8 dysregulation-mediated mitochondrial dysfunction of RVLM neurons, affecting neuronal excitability during SIH, are not fully clarified. An SIH rat model was established by administering intermittent electric foot shocks combined with noise exposure for 2 h twice daily over a period of 15 days. The impacts of PDZD8 on regulating RVLM neuronal ER stress, mitochondrial function, Apoptosis, and blood pressure (BP) of SIH rats, along with the related signaling pathway, were explored through using in-vivo and in-vitro techniques like RVLM microinjection, Western blot, flow cytometry, and immunofluorescence. We demonstrated that the ratio of c-Fos-positive Tyrosine Hydroxylase (TH) neurons, renal sympathetic nerve activity (RSNA), plasma norepinephrine (NE) levels, BP, and heart rate (HR) increased in SIH rats. The activated neuronal ER stress, impaired mitochondrial function, and Apoptosis were observed in the RVLM of SIH rats and PDZD8-deficient N2a cells. ER stress inhibitor (4-phenylbutyric acid, 4-PBA) administration effectively alleviated PDZD8 dysregulation-induced mitochondrial dysfunction and Apoptosis. Mechanistically, PDZD8 negatively regulated Calpain-2 (CAPN2) expression through modulating cytoplasmic CA²⁺ levels. In vitro, CAPN2 inhibition rescued PDZD8 deficiency-induced ER stress, mitochondrial dysfunction, and Apoptosis. In vivo, PDZD8 upregulation in the RVLM of SIH rats attenuated neuronal ER stress, mitochondrial dysfunction, and Apoptosis, thus reducing RVLM neuronal excitability, RSNA, plasma NE, BP, and HR. These effects were blocked by CAPN2 overexpression. Overall, this study revealed that PDZD8 dysregulation induced RVLM neuronal ER stress, mitochondrial damage, and Apoptosis by activating the CA²⁺-CAPN2 axis, playing a crucial pathological role in SIH progression.

Apoptosis; Ca2+-CAPN2 axis; Endoplasmic reticulum stress; Mitochondrial dysfunction; Neuronal hyperexcitation; PDZD8; Stress-induced hypertension.