Protocol for in utero injection to investigate Zika virus-related fetal microcephaly in mice

Shupeng Dong; Xiaochun Xie; Yujie Chen; Gang Long; Qiming Liang; Zhen Zhao; Jianxiong Zeng

doi:10.1016/j.xpro.2021.101057

Protocol for in utero injection to investigate Zika virus-related fetal microcephaly in mice

STAR Protoc. 2021 Dec 22;3(1):101057. doi: 10.1016/j.xpro.2021.101057. eCollection 2022 Mar 18.

Authors

Shupeng Dong^{1

2}, Xiaochun Xie^{3

4

5

6

7}, Yujie Chen⁸, Gang Long⁸, Qiming Liang^{1

2

9

10}, Zhen Zhao^{3

4}, Jianxiong Zeng^{3

4

5

6

7}

Affiliations

¹ Research Center of Translational Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200062, China.
² Shanghai Institute of Immunology, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
³ Department of Physiology and Neuroscience, University of Southern California, Los Angeles, CA 90033, USA.
⁴ Zikha Neurogenetic Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
⁵ Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences, KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research in Common Diseases, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming Yunnan 650201, China.
⁶ Kunming National High-level Biosafety Research Center for Non-Human Primates, Center for Biosafety Mega-Science, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650107, China.
⁷ National Resource Center for Non-Human Primates, National Research Facility for Phenotypic & Genetic Analysis of Model Animals (Primate Facility), Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650107, China.
⁸ Key Laboratory of Molecular Virology and Immunology, Institute Pasteur of Shanghai, Shanghai Institute for Biological Science, Chinese Academy of Sciences, Shanghai, China.
⁹ Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
¹⁰ State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200340, China.

Abstract

Zika virus (ZIKV) is linked to congenital defects including microcephaly. An infection model that can recapitulate most microcephaly-related phenotypes is crucial for understanding ZIKV pathogenesis. Here, we present a protocol to generate ZIKV from an infectious clone through a reverse genetic system and subsequently perform embryonic brain infection with the rescued ZIKV in pregnant mice. We optimized several aspects of the procedures including virus rescue and in utero injection. This protocol facilitates reproducible investigation of virus-induced cortical development defects. For complete details on the use and execution of this protocol, please refer to Zeng et al. (2020).

Keywords: Microbiology; Molecular Biology; Neuroscience; Stem Cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Body Weights and Measures
Female
Mice
Microcephaly* / etiology
Nervous System Malformations* / complications
Pregnancy
Zika Virus Infection* / congenital
Zika Virus* / genetics

Grants and funding

R01 NS110687/NS/NINDS NIH HHS/United States