Monodelphis domesticaas a fetal intra-cerebral inoculation model for Zika virus pathogenesis

Abstract

34Monodelphis domestica, also known as the laboratory opossum, is a marsupial native to South 35 America. At birth, these animals are developmentally equivalent to human embryos at 36 approximately 5 weeks of gestation which, when coupled with other characteristics including the 37 size of the animals, the development of a robust immune system during juvenile development, 38 and the relative ease of experimental manipulation, have made M. domestica a valuable model in 39 many areas of biomedical research. However, their suitability as models for infectious diseases, 40 especially diseases caused by viruses such as Zika virus (ZIKV), is currently unknown. Here, we 41 describe the replicative effects of ZIKV using a fetal intra-cerebral model of inoculation. Using 42 immunohistochemistry and in situ hybridization, we found that opossum embryos and fetuses are 43 susceptible to infection by ZIKV administered intra-cerebrally, that the infection persists long 44 term, and that the infection and viral replication consistently results in neural pathology and may 45 occasionally result in global growth restriction. These results demonstrate the utility of M. 46 domestica as a new animal model for investigating ZIKV infection in vivo. This new model will 47facilitate further inquiry into viral pathogenesis, particularly for those viruses that are 48 neurotropic, that may require a host with the ability to support sustained viral infection, and/or 49 that may require intra-cerebral inoculations of large numbers of embryos or fetuses. 50 AUTHOR SUMMARY 51Here we show that the laboratory opossum (Monodelphis domestica) is a valuable new model for 52 studying Zika virus pathogenesis. Newborns are at the developmental stage of 5-week human 53 embryos. Zika virus inoculated on a single occasion into the brains of pups at the human 54 developmental stages of 8-20 weeks post conception replicated in neuronal cells and persisted as 55 a chronic infection until the experimental endpoint at 74-days post infection. In addition, we 3 56 observed global growth restriction in one of 16 inoculated animals; global growth restriction has 57 been observed in humans and other animal models infected with Zika virus. The results illustrate 58 great potential for this new animal model for high throughput research on the neurological 59 effects of Zika virus infection of embryos and fetuses. 60 INTRODUCTION 61 Zika virus (ZIKV) is a small, enveloped positive-sense RNA virus from the family 62 Flaviviridae. Typically transmitted in a zoonotic cycle that alternates between a vertebrate host 63 and an invertebrate vector, ZIKV gained notoriety following the 2015 outbreak in Brazil, which 64 saw a dramatic increase in the number of neurological abnormalities in infants born to ZIKV-65 infected mothers [1]. Significant increases in Guillain-Barre syndrome and microcephaly during 66 this outbreak were also observed when compared to previous years [2], perhaps fulfilling the 67 theory posited by Hayes when he declared ZIKV to be neurovirulent [3...

Genetic and genomic architecture in eight strains of the laboratory opossum Monodelphis domestica

“…Marsupials are also valuable for investigating major epigenetic processes such as X-chromosome inactivation ( Hornecker et al 2007 ; Grant et al 2012 ; Rodriguez-Delgado et al 2014 ; Wang et al 2014 ; Waters et al 2018 ; Mahadevaiah et al 2020 ) and genomic imprinting ( Weidman et al 2006 ; Lawton et al 2008 ; Das et al 2012 ; Douglas et al 2014 ; Suzuki et al 2018 ). Monodelphis domestica has been used in many medical and disease studies, such as gene expression during neural development ( Dooley et al 2012 ; Sears et al 2012 ; Pavan et al 2014 ; Wheaton et al 2021 ), hypercholesterolemia and steatohepatitis ( Chan et al 2010 , 2012 ; Chan and VandeBerg 2011 ), cancer therapy and prevention ( Nair and VandeBerg 2012 ; Nair et al 2014 ), immunogenomics ( Parra et al 2008 ; Morrissey et al 2021 ; Schraven et al 2021 ), viral pathogenesis ( Thomas et al 2019 ), and influence of biological sex on social behavior, individual recognition, and associative learning ( Gil et al 2019 ).…”

CISH and IHC for the Simultaneous Detection of ZIKV RNA and Antigens in Formalin‐Fixed Paraffin‐Embedded Cell Blocks and Tissues

“…Non‐commercial probes can be quite useful for resource‐limited laboratories dedicated to routine diagnosis or research, particularly those analyzing large numbers of samples, because commercial probes (e.g., RNAscope ® , Advanced Cell Diagnostics) can be quite expensive. There are, however, only a few protocols available that use non‐commercial probes to detect ZIKV (Beaufrère et al., 2019; Thawani, Sirohi, Kuhn, & Fekete, 2018; Thomas et al., 2019). In addition, ISH is frequently combined with IHC to obtain detailed information about the relationship between ZIKV replication location and infected target cells in FFPE samples, for both diagnosis and research of this arbovirus (Calvet et al., 2016; OPS, 2018), but detailed step‐by‐step procedures are lacking.…”

Genetic and genomic architecture in eight strains of the laboratory opossumMonodelphis domestica

“…Marsupials are also valuable for investigating major epigenetic processes such as X-chromosome inactivation (Hornecker et al 2007; Grant et al 2012; Rodriguez-Delgado et al 2014; Wang et al 2014; Waters et al 2018; Mahadevaiah et al 2020) and genomic imprinting (Weidman et al 2006; Lawton et al 2008; Das et al 2012; Douglas et al 2014; Suzuki et al 2018). M. domestica has been used in many medical and disease studies, such as gene expression during neural development (Dooley et al 2012; Sears et al 2012; Pavan et al 2014; Wheaton et al 2021), hypercholesterolemia and steatohepatitis (Chan et al 2010; Chan and VandeBerg 2011; Chan et al 2012), cancer therapy and prevention (Nair and VandeBerg 2012; Nair et al 2014), immunogenomics (Parra et al 2008; Morrissey et al 2021; Schraven et al 2021), viral pathogenesis (Thomas et al 2019), and influence of biological sex on social behavior, individual recognition, and associative learning (Gil et al 2019).…”