Abstract

Background Dandy–Walker malformation (DWM) is a rare congenital brain defect whose mechanism is still not fully understood. Genetic studies have suggested that NADH Dehydrogenase 1 alpha Subcomplex 4 (NDUFA4) may be associated with DWM; however, the functional consequences of NDUFA4 dysregulation in experimental neural models remain unclear. In this study, we investigated the cellular effects of NDUFA4 deficiency, rather than aiming to establish a causal mechanism for DWM. Methods NDUFA4‐related differentially expressed proteins were analyzed using iTRAQ tandem mass spectrometry, followed by functional, pathway, and protein interaction network analyses. Then, voltage‐dependent anion Channel 1 (VDAC1), apoptosis‐, endoplasmic reticulum (ER) stress–related proteins, and ETC Complex IV activity were assessed in NDUFA4 knockout mice. After NDUFA4 and VDAC1 knockdown, cell proliferation, apoptosis, mitochondrial status, and ER function were evaluated by CCK‐8, Edu staining, flow cytometry, and Western blot in C8‐D1A cells. In addition, the interaction between NDUFA4 and VDAC1 was evaluated using immunofluorescence and coimmunoprecipitation. Results NDUFA4 knockout upregulated VDAC1, ER stress‐ and apoptosis‐related proteins, and inhibited ETC complex IV activity in mice. NDUFA4 knockdown inhibited proliferation and promoted apoptosis, ER stress, and mitochondrial damage in C8‐D1A cells, and these changes were partially reversed by VDAC1 knockdown. Moreover, NDUFA4 and VDAC1 colocalized in C8‐D1A cells and mouse cerebellar tissue, and NDUFA4 was found to interact with VDAC1. Conclusions NDUFA4 deletion was associated with VDAC1 upregulation, mitochondrial damage, ER stress, and apoptosis‐related changes in our experimental models. These findings may provide insight into cellular changes potentially relevant to DWM; however, they do not establish a direct causal relationship.