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Male offspring born to mildly ZIKV-infected mice are at risk of developing neurocognitive disorders in adulthood

Abstract

Congenital Zika virus (ZIKV) syndrome may cause fetal microcephaly in ~1% of affected newborns. Here, we investigate whether the majority of clinically inapparent newborns might suffer from long-term health impairments not readily visible at birth. Infection of immunocompetent pregnant mice with high-dose ZIKV caused severe offspring phenotypes, such as fetal death, as expected. By contrast, low-dose (LD) maternal ZIKV infection resulted in reduced fetal birth weight but no other obvious phenotypes. Male offspring born to LD ZIKV-infected mothers had increased testosterone (TST) levels and were less likely to survive in utero infection compared to their female littermates. Males also presented an increased number of immature neurons in apical and basal hippocampal dendrites, while female offspring had immature neurons in basal dendrites only. Moreover, male offspring with high but not very high (storm) TST levels were more likely to suffer from learning and memory impairments compared to females. Future studies are required to understand the impact of TST on neuropathological and neurocognitive impairments in later life. In summary, increased sex-specific vigilance is required in countries with high ZIKV prevalence, where impaired neurodevelopment may be camouflaged by a healthy appearance at birth.

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Fig. 1: Maternal ZIKV pathogenesis in immunocompetent allogeneic pregnant mice.
Fig. 2: Vertical ZIKV transmission.
Fig. 3: Reproductive outcome.
Fig. 4: TST levels of pregnant women and newborns.
Fig. 5: Apical and basal dendrites of adult male and female offspring.
Fig. 6: Assessment of spatial learning and memory of adult male and female offspring.

Data availability

Accession numbers of the ZIKV strains used to perform the phylogenetic analysis are indicated in Supplementary Fig. 1. The data that support the findings of this study are available from the corresponding author on request.

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Acknowledgements

The Heinrich Pette Institute, Leibniz Institute for Experimental Virology is supported by the Free and Hanseatic City of Hamburg and the Federal Ministry of Health. This study was supported by the Federal Ministry of Health (G.G.), the German Research Center for Infection (DZIF) (G.G.), the Niedersachsen-Research Network on Neuroinfectiology (N-RENNT) of the Ministry of Science and Culture of Lower Saxony, Germany (W.B., W.L.) and the German Federal Ministry of Education and Research (Infrafrontier grant 01KX1012) (M.H.A.). F.C.d.A. is supported by Deutsche Forschungsgemeinschaft (DFG) Grant (FOR 2419, CA1495/1-1 and CA 1495/4-1), ERA-NET Neuron Grant (Bundesministerium für Bildung und Forschung, BMBF, 01EW1410 ZMNH AN B1), Landesforschungsförderung Hamburg (Z-AN LF) and University Medical Center Hamburg-Eppendorf (UKE). R.B. is supported by the Deutsche Forschungsgemeinschaft (BA 1505/8-1). We are grateful for the excellent technical contribution of all staff at the technology platform small animal models of the Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg and the technical staff of the Department of Pathology, University of Veterinary Medicine, Hannover. We thank T. Andreas from the Department of Obstetrics and Fetal Medicine at the University Medical Center Hamburg-Eppendorf for his excellent technical support with the preparation of pregnant mice. We thank P. Pruunsild and H. Bading, Department of Neurobiology, Interdisciplinary Center for Neurosciences at Heidelberg University for providing human NPCs. We thank U. Markert, Department of Obstetrics, Placenta-Lab, University Hospital Jena for providing the human placental cells.

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G.G. conceived the study. S.S.-B. and G.G. designed and coordinated the experiments. S.S.-B., K.W.-G., C.D., A.P., G.P.-S., U.M. and S.Thi. performed all infection studies in mice. TST treatment of mice was performed by H.L., J.S. and S.H. S.Tha., I.A.A., T.S., N.M.K., C.D. and K.W.-G. performed the qRT–PCR assays. S.Tha., C.D., K.W.-G. and A.P. performed the hormone ELISA as well as the cytokine assays. T.R. measured the TST levels. V.H., W.B., V.M.P. and I.G. performed the histological and immunohistological examinations, TUNEL and cytokeratin staining, and the in situ hybridization and analysis. B.S. performed the Iba1 staining and respective analysis of fetal brains. H.I. performed the MRI scans on the brains of the offspring. M.R. performed Golgi staining and assisted with spine analysis. R.S. performed image acquisition, and the dendritic and spine analyses. F.C.d.A. coordinated the brain analysis. U.B. performed the immunohistochemical analyses of retinas. T.M. and R.B. performed the ZIKV replication kinetics in cell culture. Histopathological findings were analysed and discussed by P.A., M.A.F., V.H., I.G., V.M.P. and W.B. S.J. and T.S. measured viral IgG and IgM titres. D.C. performed the phylogenetic analysis. S.M.H., O.A., F.M., V.K., R.D., L.S., W.L., I.W. and C.K. performed and/or analysed the behavioural experiments. H.F. and M.H.d.A. coordinated and conceived the animal phenotypic tests. S.B. and L.R. were responsible for the recruitment of the mother–child cohort in Iquitos, Peru and for the qRT–PCR analysis of patient sera. J.S.-C., O.V. and M.G. provided material, analysed the data and discussed the study. S.S.-B. and G.G. wrote the manuscript. All authors revised the manuscript.

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Correspondence to Gülsah Gabriel.

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Stanelle-Bertram, S., Walendy-Gnirß, K., Speiseder, T. et al. Male offspring born to mildly ZIKV-infected mice are at risk of developing neurocognitive disorders in adulthood. Nat Microbiol 3, 1161–1174 (2018). https://doi.org/10.1038/s41564-018-0236-1

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