TY  - EJOU
AU  - Latino, Debora 
AU  - Falvo, Sara 
AU  - Grillo, Giulia 
AU  - Santillo, Alessandra 
AU  - Baccari, Gabriella Chieffi 
AU  - Venditti, Massimo 
AU  - Fiore, Maria Maddalena Di 

TI  - Mitochondria-Associated Endoplasmic Reticulum Membranes (MAMs) and Mitochondrial Dynamics as a Target of D-Aspartate in TM3 Leydig Cells
T2  - BIOCELL

PY  - 
VL  - 
IS  - 
SN  - 1667-5746

AB  - <b>Objectives:</b> Mitochondria-associated endoplasmic reticulum membranes (MAMs) are key communication sites between the endoplasmic reticulum (ER) and mitochondria, coordinating lipid transfer, calcium signaling, mitochondrial dynamics, and cellular homeostasis. This study investigated the effects of D-aspartate (D-Asp) on MAMs function and mitochondrial quality in TM3 Leydig cells. <b>Methods:</b> Murine TM3 cells were treated with 100 μM D-Asp for 24 h. Protein expression of MAM- and mitochondrial-related markers was assessed by Western blotting, while mitochondrial membrane potential (MMP) was evaluated using the TMRE assay. <b>Results:</b> D-Asp significantly increased ATAD3A (<i>p</i> = 0.018) and SOAT1 (<i>p</i> = 0.023), markers of lipid transfer, and GRP75 (<i>p</i> = 0.011) and VDAC (<i>p</i> = 0.021), involved in calcium signaling, without significantly affecting the ER stress marker GRP78/BiP. D-Asp also enhanced mitochondrial biogenesis and fusion, increasing NRF1 (<i>p</i> = 0.015), TFAM (<i>p</i> = 0.010), MFN1 (<i>p</i> = 0.019), MFN2 (<i>p</i> = 0.03), and OPA1 (<i>p</i> = 0.05) expression, together with TOMM20 (<i>p</i> = 0.049), indicating increased mitochondrial mass. Conversely, DRP1 expression, a marker of mitochondrial fission, was reduced (<i>p</i> = 0.011). Increased TMRE fluorescence confirmed improved mitochondrial membrane potential and functionality. In addition, D-Asp decreased Bax and cytochrome c expression, suggesting reduced apoptotic signaling. <b>Conclusion:</b> D-Asp directly modulates MAM-related processes and mitochondrial homeostasis in TM3 Leydig cells. It promotes lipid trafficking and calcium signaling, enhances mitochondrial biogenesis, fusion, mass, and membrane potential, reduces mitochondrial fission, and attenuates apoptosis. These findings support a broad regulatory role for D-Asp in maintaining mitochondrial function and cellular homeostasis in Leydig cells.
KW  - Leydig cells; mitochondria; endoplasmic reticulum; mitochondria-associated membranes; lipid transfer; calcium signaling; mitochondrial dynamics; mitochondrial membrane potential; apoptosis

DO  - 10.32604/biocell.2026.087384