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Sex Differences in Early Postnatal Microglial Colonization of the Developing Rat Hippocampus Following a Single-Day Alcohol Exposure

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Abstract

Microglia are involved in various homeostatic processes in the brain, including phagocytosis, apoptosis, and synaptic pruning. Sex differences in microglia colonization of the developing brain have been reported, but have not been established following alcohol insult. Developmental alcohol exposure represents a neuroimmune challenge that may contribute to cognitive dysfunction prevalent in humans with Fetal Alcohol Spectrum Disorders (FASD) and in rodent models of FASD. Most studies have investigated neuroimmune activation following adult alcohol exposure or following multiple exposures. The current study uses a single day binge alcohol exposure model (postnatal day [PD] 4) to examine sex differences in the neuroimmune response in the developing rat hippocampus on PD5 and 8. The neuroimmune response was evaluated through measurement of microglial number and cytokine gene expression at both time points. Male pups had higher microglial number compared to females in many hippocampal subregions on PD5, but this difference disappeared by PD8, unless exposed to alcohol. Expression of pro-inflammatory marker CD11b was higher on PD5 in alcohol-exposed (AE) females compared to AE males. After alcohol exposure, C-C motif chemokine ligand 4 (CCL4) was significantly increased in female AE pups on PD5 and PD8. Tumor necrosis factor-α (TNF-α) levels were also upregulated by AE in males on PD8. The results demonstrate a clear difference between the male and female neuroimmune response to an AE challenge, which also occurs in a time-dependent manner. These findings are significant as they add to our knowledge of specific sex-dependent effects of alcohol exposure on microglia within the developing brain.

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Acknowledgements

The authors would like to thank Julia Johansson and Shaqran Shareeq for assistance with immunohistochemistry, and all of the undergraduate students for their help with animal generation and care. This work was supported by the National Institutes of Health/NIGMS COBRE: The Delaware Center for Neuroscience Research IP20GM103653-01A1 grant to AYK.

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  1. Department of Psychological and Brain Sciences, University of Delaware, 108 Wolf Hall, Newark, DE, 19716, USA

    M. J. Ruggiero, K. E. Boschen, T. L. Roth & A. Y. Klintsova

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  1. M. J. Ruggiero
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  2. K. E. Boschen
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  3. T. L. Roth
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  4. A. Y. Klintsova
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Correspondence to A. Y. Klintsova.

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Online Resource 1

Estimated number of microglia in hippocampal subregions of females on PD5 and PD8 following PD4 alcohol exposure. Cell counts were conducted in the following the hippocampal subregions: A) dentate gyrus molecular layer, B) granule cell layer (GCL), and C) hilus; D) CA1 stratum oriens, E) pyramidal cell layer (PCL), and F) stratum radiatum; G) CA3 stratum oriens, H) PCL, and I) stratum radiatum. Main effects of day are indicated with lines/asterisks and main effects of treatment are indicated by the asterisks directly over the bars. ** = p < .01, *** = p < 0.001, **** = p < 0.0001. SC = suckle control, SI = sham-intubated, AE = alcohol-exposed. All data are expressed as mean ± SEM. (GIF 160 kb)

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Online Resource 2

Estimated number of microglia in hippocampal subregions of male rats on PD5 and PD8 following PD4 alcohol exposure. Cell counts were conducted in the following the hippocampal subregions: A) dentate gyrus molecular layer, B) granule cell layer (GCL), and C) hilus; D) CA1 stratum oriens, E) pyramidal cell layer (PCL), and F) stratum radiatum; G) CA3 stratum oriens, H) PCL, and I) stratum radiatum. Main effects of day are indicated with lines/asterisks. **** = p < 0.0001. SC = suckle control, SI = sham-intubated, AE = alcohol-exposed. All data are expressed as mean ± SEM. (GIF 173 kb)

High Resolution Image (TIFF 390 kb)

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Ruggiero, M.J., Boschen, K.E., Roth, T.L. et al. Sex Differences in Early Postnatal Microglial Colonization of the Developing Rat Hippocampus Following a Single-Day Alcohol Exposure. J Neuroimmune Pharmacol 13, 189–203 (2018). https://doi.org/10.1007/s11481-017-9774-1

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