Fig. 6: Mitochondria-nuclear communication at fertilization determines ICM telomere length.
From: Telomere length in offspring is determined by mitochondrial-nuclear communication at fertilization
Ovulated eggs from control or rotenone-exposed females underwent in vitro fertilization and embryo culture in media containing 10 µM BGP-15, 50 nM MitoQ, or 1 nM SS-31 (+) or media with the equivalent volume of vehicle (-) for 8 h and zygotes were then cultured in standard (untreated) media to the blastocyst stage (a) Created in BioRender. Gordon, Y. (2025) https://BioRender.com/y04d797. At 6 h post-fertilization, zygotes were labelled with MitoSox Red superoxide (mtROS) indicator (red, and DNA stain Hoechst-3342; blue) and red fluorescence measured (b: n = 31 control, n = 29 rotenone, n = 18 rotenone+BGP-15; d: n = 19 control, n = 13 control+MitoQ, n = 13 rotenone, n = 14 rotenone+MitoQ; f: n = 12 control, n = 12 control+SS-31, n = 16 rotenone, n = 18 rotenone+SS-31). At the blastocyst stage ICM telomere length (telomere/ Rn18s (2-ΔΔCt)) was measured by qPCR (c: n = 35 control, n = 31 control+BGP-15, n = 18 rotenone, n = 30 rotenone+BGP-15; e: n = 24 control, n = 19 control+MitoQ, n = 21 rotenone, n = 18 rotenone+MitoQ; g: n = 19 control, n = 26 control+SS-31, n = 16 rotenone, n = 19 rotenone+SS-31). Mice were exposed to rotenone (150 ppm in chow) for three weeks before hormone stimulation, mating and zygote collection. Pronuclei were transferred between zygotes derived from control (cont) or rotenone-exposed (rote) females in each possible combination, and reconstructed embryos cultured to blastocyst (h) Created in BioRender. Gordon, Y. (2025) https://BioRender.com/h97n093. ICMs from blastocysts were analyzed for telomere length (telomere/ Rn18s (2-ΔΔCt)) (i; n = 33 cont-cont, n = 24 rote-cont, n = 27 cont-rote, n = 28 rote-rote). Individual data points are plotted, horizontal lines are mean ± SEM (b, d, f). Violin plots show population distribution, and horizontal lines are mean ± SEM (c, e, g, i). qPCR data was log transformed for statistical analysis via two-way ANOVA (c, e, g) or one-way ANOVA (i), MitoSox Red analyzed via one-way ANOVA (b, d, f); *p < 0.05, **p < 0.008, ***p ≤ 0.0004, ****p < 0.0001. For exact P values see Supplementary Table 6. Representative images shown, scale bar: 20 µm (b, d, f). Source data are provided as a Source Data File.
