Scale bars, 10 μm (main images; top) and 5 μm (magnified views of the boxed regions; middle and bottom). Arrowheads indicate tMac-derived mitoD2+ mitochondria within LCs. Following sorting from WT mice of different ages, LCs were co-cultured with MHCIIhi tMacs from three-month-old Cx3cr1CreER; R26mitoD2 (mitoD2) mice treated with TAM. Scale bars, 15 μm (main images; top) and 10 μm (magnified views of the boxed regions; middle and bottom). Accordingly, knockout of PGC-1α in mice dramatically increases sensitivity to oxidative stress-induced damage in SN dopaminergic cells and hippocampal neurons, while increasing PGC-1α levels protects cultured neurons from oxidative stress-mediated death . Our study showed that testosterone supplementation increased PGC-1α expression and stimulated the transcription, translation, and functional activity of GSH-PX, CAT, and Mn-SOD in the aged male rat brain. Both physiological (e.g., age-related) and therapeutically induced (e.g., hormone replacement) alterations in circulating hormone levels have shown to influence changes in mitochondrial function 16, 18, 33, 34. Leydig cells in the testes utilize ATP produced by mitochondria to create testosterone. Nutrition plays a significant role as well, with nutrients like vitamin D and omega-3 fatty acids positively influencing mitochondrial function and testosterone production. Factors such as oxidative stress and mitochondrial dysfunction can negatively affect testosterone levels. Now you know mitochondria not only power your cells but are also essential for hormone production, especially testosterone. However, these should always be used with caution and under medical supervision, as they can affect mitochondrial gene expression and protein levels. Exercise isn't only good for your muscles, it's also an effective way to promote mitochondrial biogenesis (the creation of new mitochondria) and improve testosterone levels. Conversely, LCs acquire EVs containing functional mitochondria from MHCIIhi tMacs via ITGβ1–VCAM1 interactions. Second, although mitochondria were identified as key functional components, both EV populations contain additional cargos whose contributions cannot be excluded. Although this study highlights the critical role of the mitochondrial transfer network between LCs and tMacs, two important questions remain unresolved. However, we cannot exclude the possibility that alternative mechanisms may also contribute to the observed mitochondrial transfer network between LCs and tMacs. Deletion of Vcam1 in LCs or treatment with a VCAM1-blocking antibody significantly impaired mitochondrial uptake, suggesting that VCAM1 confers specificity for the recognition of tMac-EVs. Interestingly, VCAM1 is also recognized as a marker of LCs, although its specific role in these cells has not been well defined52. VCAM1 is traditionally known to be expressed on the surface of endothelial cells where it mediates cell adhesion by interacting with adhesion molecules on target cells51. Meanwhile, compared to 6Mon animals, the activities of mitochondrial complexes I and IV in the HIPP showed a slight, non-significant reduction in 24Mon control rats. These findings demonstrate that TP supplementation attenuates neuronal dysfunction and neuronal loss in the SN and HIPP of aged male rats. TP supplementation increases the expression of TH and DAT in the SN of aged male rats. Uptake of EVs has been reported to occur via endocytosis or direct fusion with the acceptor cell membrane39. Consistent with this, EV-contained mitochondria exhibited markedly lower MitoSOX signals (Fig. 5q,r). Functionally, EV-encapsulated mitochondria maintained a responsive MMP, depolarizing with FCCP and hyperpolarizing with oligomycin (Fig. 5l). Staining with MitoTracker Red (MTR) revealed that the majority (77.37 ± 1.92%) of tMac-EVs contained mitochondria (Fig. 5i). Among the 1,057 identified proteins, almost 60% were ascribed as components of mitochondrion (26.35%), plasma membrane (25.99%) and cytoplasmic vesicle (6.56%; Fig. 5f). M,o,q, Scale bars, 3 μm (left) and 2 μm (right; magnified views of the boxed regions).