Cells were loaded with the dye at a concentration of 0. Indeed, brain glucose hypometabolism has been observed in AD patients well before the onset of clinical symptoms [ 16 ]. In experiments using differentiated cells, well plates were coated with 0. Mitochondrial respiration The investigation of mitochondrial respiration and cellular glycolysis was performed using the Seahorse Bioscience XF24 analyser. Mounting evidence suggests that mitochondrial dysfunction serves as a catalyst in AD, since the disease is associated with a decline in bioenergetic activity and an increase in oxidative stress that can already be detected at early disease stage in both FAD and SAD [ 10 , 13 — 15 ].
Mitochondrial respiration The investigation of mitochondrial respiration and cellular glycolysis was performed using the Seahorse Bioscience XF24 analyser. The PL cells presented a decreased maximal respiration and spare respiratory capacity, while APP cells exhibited, in addition, a decrease in basal respiration, ATP turnover, and glycolytic reserve. Our findings lend further support to the neuroprotective effects of neurosteroids in AD and may open new avenues for the development of gender-specific therapeutic approaches in AD. For statistical comparisons of two groups, student unpaired t tests were used. Data were extracted from the Seahorse XF software, and bioenergetic parameters basal respiration, ATP turnover, maximal respiration, spare respiratory capacity, and glycolytic reserve were calculated according to the guideline of the company. In , Baulieu and colleagues characterized a new category of steroids that is synthesized in the nervous system and persists in substantial amounts after removal of the peripheral steroidogenic glands [ 20 ]. Transmembrane distribution of the dye is dependent on MMP. Stably transfected cell clones were selected with hygromycin. In line with that study, a growing body of evidence attests to the neuroprotective effects of neurosteroids, especially estrogenic compounds, against AD-related cellular injury reviewed in [ 18 ]. On the basis of our previous findings, it was therefore of interest to examine whether neurosteroids are able to alleviate mitochondrial deficits manifested in these AD cellular models. Cells were plated in 5 replicates into a white well cell culture plate at a density of 1. Electronic supplementary material The online version of this article doi: The bioluminescent method measures the formation of light from ATP and luciferin by luciferase. Of note, both AD cell culture models, APP and PL cells, express a variety of neuronal receptors, including steroid receptors progesterone, estrogen, and androgen receptors and exhibit the characteristics of a mitochondrial malfunction when compared to their respective controls [ 26 — 29 ]. Cells were loaded with the dye at a concentration of 0. Mounting evidence suggests that mitochondrial dysfunction serves as a catalyst in AD, since the disease is associated with a decline in bioenergetic activity and an increase in oxidative stress that can already be detected at early disease stage in both FAD and SAD [ 10 , 13 — 15 ]. In experiments using differentiated cells, well plates were coated with 0. Biliouris , Undine E. However, little is known about the influence of neurosteroids on AD-related mitochondrial dysfunction. Testosterone was from AppliChem Darmstadt, Germany. In particular, their impact on ATP production, mitochondrial membrane potential MMP , mitochondrial respiration, and glycolysis was investigated. They shared decreased ATP levels as well as impaired mitochondrial respiration [ 26 , 27 ] but differed in the underlying mechanisms. We show that while APP and PL cells both display a drop in ATP levels, they present distinct mitochondrial impairments with regard to their bioenergetic profiles. Determination of mitochondrial membrane potential The MMP was measured using the fluorescent dye tetramethylrhodamine, methyl ester, perchlorate TMRM [ 19 ]. Interestingly, no mutations in the tau encoding gene have been identified so far in FAD.
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