Transient, early, female-specific increase in cortical glial fibrillary acidic protein distribution in the Syrian hamster model of mild peripheral COVID-19

Abstract

BackgroundMild-moderate respiratory COVID-19 is commonly associated with a range of neurological symptoms. The mechanisms linking this peripheral disease to cognitive symptoms are thought to include heightened circulating cytokines and other inflammatory mediators resulting in a leaky blood-brain barrier and increased neuroinflammation (i.e., inflammation taking place in the brain). This can lead to aberrant synaptic transmission and cognitive dysfunction. A key component of neuroinflammation is the reactivity of astrocytes, in a process termed astrogliosis, associated with altered morphology, proliferative capacity, gene expression, and function. Accumulating evidence suggests astrogliosis likely occurs in mild-moderate COVID-19; however, there has been limited investigation. In this study, we quantified changes to astrocytes in a Syrian hamster model of mild-moderate respiratory COVID-19. MethodsWe used an intranasal inoculation model to produce mild-moderate respiratory COVID-19 in 8-10-week-old male and female Syrian hamsters. We extracted brains at 1-, 3-, 5-, 7-, and 31-days post-inoculation and from uninfected controls, and immunolabelled brain sections with astrocyte- (GFAP and SOX9) and neuron-specific (NEUN) markers. We captured tiled confocal micrographs of entire brain sections and analyzed the resulting signals from five regions of interest: cortex, corpus callosum, hippocampus, third ventricle, and dorsal striatum. ResultsTo systematically quantify cell-type-specific labelling for astrogliosis markers, we first developed an unbiased pipeline. We found a transient increase in GFAP signal density in female hamster, specifically in the cortex at 3 days post-inoculation. There were no corresponding changes noted in astrocyte (SOX9), neuron (NEUN) or total cell (Hoechst) numbers. Moreover, there were no changes in male hamsters at any timepoint in any region of interest. ConclusionsOur findings provide the first spatiotemporal insight into astrogliosis in a hamster model of mild-moderate respiratory COVID-19. We identified a transient and sex-specific increase in GFAP signal density, indicative of astrogliosis. Our findings contribute to the literature surrounding sex differences in (neuro)immune responses and add to the growing body of COVID-19 literature, in which sex-specific outcomes are apparent in both human patient populations and rodent experimental models.