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Through TEM analysis of testes, we identified mitochondria within the EVs with cristae area fractions comparable to mitochondria in tMacs (Fig. 5j,k). A comparison of the tMac-EV proteome with the MitoCarta3.0 and Vesiclepedia databases24,25 revealed 183 and 864 overlapping proteins, respectively (Fig. 5g). G, Venn diagrams of the tMac-EV proteome with proteins reported in the MitoCarta3.0 and Vesiclepedia database. We purified GFP+ particles devoid of nuclei from the testes of Cx3cr1GFP mice (Extended Data Fig. 8a–c). Following the administration of hCG to induce elevated testosterone levels, we observed a noticeable increase in the number of tdTomato+ LCs with GFP+ particles (Extended Data Fig. 7c–g). To confirm the transfer of tMac-derived particles to LCs, we collected testes from Cyp17a1Cre; R26tdTomato; Cx3cr1GFP mice for analysis.
For imaging at ×25 effective magnification, each sample was scanned by three-tile light sheets axially at a 3.5-μm Z-step size to image the whole sample. For imaging at ×6.3 effective magnification, each sample was scanned by one-tile light sheets axially at a 3.5-μm Z-step size to image the whole sample. Briefly, the PFA-fixed testes were immersed in CUBIC-L solution with shaking at 37 °C for three days. The testes were dissected and fixed overnight in 4% PFA at 4 °C, followed by extensive washing with PBS as described previously65.
Use red light therapy, which has been shown to support mitochondrial function and enhance testosterone production by stimulating ATP production in cells. Supplements of testosterone propionate to castrated male rats ameliorated the activity of mitochondrial complex I and upregulated the expression of mitochondrial ND1 and ND4. In conclusion, our study revealed that testosterone supplementation improved exploratory behavior, attenuated neuronal dysfunction and neuronal loss, and ameliorated mitochondrial dysfunction by enhancing both mitochondrial antioxidative capacity and mitochondrial biogenesis of aged male rats. Increased PINK1/Parkin and decreased P62 expression in the SN and HIPP of testosterone-supplemented rats further suggested that enhanced mitophagy likely contributes to the beneficial effect of testosterone supplementation against mitochondrial dysfunction in the aged rat brain. These findings, along with those of the present study, demonstrated that cognitive/behavioral deficits and mitochondrial dysfunction in the aged male brain are, to some extent, related to decreased serum testosterone levels. In turn, orchiectomy was shown to disturb mitochondrial function, evidenced by increased mitochondrial H2O2 production and decreased MMP in the SN of adult male rats . These findings strongly suggest that testosterone supplementation ameliorates age-related brain mitochondria dysfunction in male rats by enhancing both mitochondrial antioxidative capacity and mitochondrial biogenesis.
Enhanced mitochondrial activity correlates with increased testosterone levels, improving physical performance and mental clarity. Cultivating mitochondrial health provides a foundation for optimizing testosterone levels, which can benefit both athletic performance and overall quality of life. Addressing mitochondrial health can optimize testosterone levels, bolstering physical strength, and mental clarity.
Arrows indicate tMac-derived mitoD2+ mitochondria within tdTomato+ LCs. Confocal microscopy revealed that tdTomato+ host LCs contained significantly more mitoD2+ mitochondria from MHCIIhi tMacs than from CD206hi tMacs (Fig. 7c,d). Unexpectedly, confocal analysis revealed that significantly fewer CD206hi tMacs transferred mitoD2+ mitochondria to LCs compared with MHCIIhi tMacs (Fig. 7a,b). To test this, we sorted CD206hi and MHCIIhi tMacs by FACS from TAM-treated adult Cx3cr1CreER; R26mitoD2 mice and co-cultured them with tdTomato+ LCs in vitro (Fig. 7a,b and Extended Data Fig. 9d). By contrast, antimycin A-treated tMac-EVs failed to augment testosterone or ATP (Fig. 6m,n). Q, ATP levels of LCs from the three groups. P, Testosterone production of LCs from the three groups at 6 h.
TREM2 has been shown to play a key role in macrophages by mediating the removal of dysfunctional subcellular fragments in various tissues36,37. L, Representative images of the testes of control and cKO mice nine days after mitoD2+ LC transplantation. Collectively, these data indicate that LCs preferentially transfer mitochondria to CD206hi tMacs. High-resolution imaging analysis of Cyp17a1Cre; R26mitoD2 testes confirmed that LC-derived mitochondria were predominantly found in CD206hi tMacs, with negligible uptake by MHCIIhi tMacs (Fig. 4c,d). - http://111.230.243.127:3000/lisasugerman77
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