Research Paper Volume 11, Issue 16 pp 6336—6357

Inhibition of de novo ceramide biosynthesis affects aging phenotype in an in vitro model of neuronal senescence

Figure 1. Effects of aging and L-CS on resting calcium (Ca2+) levels in cortical neurons.(A) The pictogram illustrates the experimental paradigm employed in the study. (B) Representative brightfield micrographs of control and aged neuronal cultures treated either with L-CS or vehicle (scale bar 100 µm). Please, note that aged cultures are devoid of signs of neuronal death. (C) Bar graphs depict the relative abundance of ceramides in vehicle- and L-CS-treated aged neurons (n=3 for both conditions). (D) Representative fluorescent micrograph of a fura-2-loaded cultured cortical neuron (the image reports dye emission when excited at 380 nm, scale bar 25 µm). (E) Bar graphs depict dendritic Ca2+levels of vehicle- or L-CS-treated control neurons (vehicle: n=102 proximal and n=85 distal dendrites from 43 neurons; L-CS: n=115 proximal and n=84 distal dendrites from 38 neurons; p>0.05). (F) Bar graphs depict dendritic Ca2+levels of vehicle- or L-CS-treated aged neurons (vehicle: n=182 proximal and n=156 distal dendrites from 40 neurons; L-CS: n=177 proximal and n=155 distal dendrites from 44 neurons; p>0.05). (G) Bar graphs depict somatic Ca2+levels of vehicle- or L-CS-treated control and aged neurons (ControlVeh: n=1357 cells and ControlL-CSn=1015; AgedVehn=539 cells and AgedL-CSn=497 cells obtained from 10-23 independent experiments). In C and E-F means were compared by unpaired Student t-test. In G means were compared by two-way ANOVA followed by Tukey post-hoc test. * indicates p<0.05, *** p<0.001.