Fig. 3: Intracytoplasmic sperm injection (ICSI) of sperm incubated with extracellular vesicles (EVs) secreted following stress recovery alter offspring neurodevelopment. | Nature Communications

Fig. 3: Intracytoplasmic sperm injection (ICSI) of sperm incubated with extracellular vesicles (EVs) secreted following stress recovery alter offspring neurodevelopment.

From: Reproductive tract extracellular vesicles are sufficient to transmit intergenerational stress and program neurodevelopment

Fig. 3

a Schematic representation of ICSI paradigm using sperm incubated with vehicle (EVVeh) or corticosterone-treated (EVCort) EVs, followed by microinjection into super-ovulated oocytes obtained from the same female donors. Zygotes from both EV-treatment groups were transferred into the designated right or left side of the same naive foster females to assess offspring neurodevelopment at mid-gestation (E12.5, top), or into separate recipient females for assessment of an F1 stress phenotype in adulthood (bottom). b, c Gene set enrichment analysis (GSEA) was used to analyze RNA-sequencing of E12.5 brains, with significant enrichment of gene sets related to (b) synaptic signaling and (c) neurotransmitter transport in embryos from EVCort sperm compared with EVVeh sperm (N = 6 embryos/EV treatment, GSEA normalized enrichment score (NES) > |1.8|, FDR < 0.05). d The top three significant clusters of gene ontology (GO) terms enriched in EVCort E12.5 brains determined by GSEA and clustered under parent terms were related to synaptic signaling (N = 6 embryos/treatment, NES > |1.8| and FDR < 0.05). e EVCort adult F1 offspring showed the same phenotypic pattern of stress reactivity to an acute 15-min restraint (gray bar) as paternal stress F1 offspring, compared with EVVeh F1 offspring (two-way rmANOVA, interaction of EV treatment × time F(3,21) = 8.480, p = 0.0007, time (F(3,21) = 122.6, p = 1.8125−13), EV treatment (F(1,7) = 1.868, p = 0.2139, N = 4–5 offspring/EV treatment), with a significant reduction in the maximal rise of corticosterone at 30-min (Bonferroni’s post hoc test, t(28) = 2.733, *adjusted p = 0.043). Error bars represent mean ± SEM.

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