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bioRxiv · 10.1101/2025.04.04.647329

Haploinsufficiency of eNOS mitigates beneficial effects of maternal exercise on fetal heart development during pregestational diabetes

Van Neck, R.·Lu, X.·Wisse, L. J.·DeRuiter, M. C.·Feng, Q.
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Abstract

BackgroundPregestational diabetes (PGD) increases congenital heart defect (CHD) risk over five-fold. Maternal exercise enhances endothelial nitric oxide synthase (eNOS) activity, benefiting embryos, though its causal role remains unclear. This study investigated eNOSs role in maternal exercise-mediated protection of fetal heart development in a PGD mouse model. MethodsPGD was induced in eNOS+/- or wild-type (WT) female mice via streptozotocin before breeding with WT or eNOS+/- males. Pregnant females had access to a running wheel for voluntary exercise or remained sedentary. Fetuses were collected at embryonic day (E) 18.5 for genotyping and CHD assessment. E12.5 hearts were analyzed for proliferation, apoptosis, oxidative stress, and eNOS protein levels. ResultsMaternal exercise normalized litter size and mortality rates in offspring of diabetic eNOS+/- females but did not reduce CHD incidence in offspring of WT or eNOS+/- females with PGD. CHDs included septal defects, double outlet right ventricle, and valve defects. Exercise increased coronary artery density but not capillary density. Proliferation deficits at E12.5 were restored by exercise, yet oxidative stress remained elevated. Maternal exercise in eNOS+/- dams during PGD did not significantly change eNOS protein levels in both eNOS+/+ and eNOS+/- fetal hearts. Offspring genotype did not impact CHD incidence, cell proliferation, apoptosis or oxidative stress. ConclusionsMaternal exercise does not prevent CHDs in PGD offspring of eNOS+/- mice. Its ability to mitigate PGD-induced oxidative stress is eNOS-dependent and essential for improving heart morphology. Graphic Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=178 SRC="FIGDIR/small/647329v1_ufig1.gif" ALT="Figure 1"> View larger version (42K): org.highwire.dtl.DTLVardef@4c9e4forg.highwire.dtl.DTLVardef@2d60fforg.highwire.dtl.DTLVardef@13b2d72org.highwire.dtl.DTLVardef@249a0c_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Van Neck, R., Lu, X., Wisse, L. J., DeRuiter, M. C., Feng, Q.. 2025-04-05. Haploinsufficiency of eNOS mitigates beneficial effects of maternal exercise on fetal heart development during pregestational diabetes. https://doi.org/10.1101/2025.04.04.647329

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