Abstract
Microglia are implicated in multiple neurodegenerative disorders, many of which display sexual dimorphisms and have symptom onsets at different ages. P2 purinergic receptors are critical for regulating various microglial functions, but little is known about how their expression varies with age or sex. Therefore, comprehensive information about purinergic receptor expression in normal microglia, in both sexes, over age is necessary if we are to better understand their roles in the healthy and diseased CNS. We analyzed the expression of all fourteen rodent P2X and P2Y receptors in CD11b+ cells freshly-isolated from the brains of C57Bl/6 mice at five different ages ranging from postnatal day 3 to 12 months, in males and females, using quantitative RT-PCR. We also compared purinergic receptor expression in microglia freshly-isolated from 3 day-old pups to that in primary neonatal microglial cultures created from mice of the same age. We observed patterns in P2 receptor expression with age, most notably increased expression with age and age-restricted expression. There were also several receptors that showed sexually dimorphic expression. Lastly, we noted that in vitro culturing of neonatal microglia greatly changed their P2 receptor expression profiles. These data represent the first complete and systematic report of changes in purinergic receptor expression of microglia with age and sex, and provide important information necessary for accurate in vitro modeling of healthy animals.
Highlights
Microglia are the primary resident immune cell population in the central nervous system (CNS)
We evaluated the expression of all rodent P2Rs in freshly-isolated mouse microglia, and we display them here in groups according to their overall expression patterns
P2X1 and P2X4 expression varies over age We found that mRNA expression of both P2X1 (Figure 2A) and P2X4 (Figure 2B) vary with age
Summary
Microglia are the primary resident immune cell population in the central nervous system (CNS). They phagocytose debris following neuronal remodeling processes, help maintain CNS integrity, and perform neuronal support functions through the production of neurotrophins and growth factors [1]. Uncontrolled microglial activation and their resulting production of neurotoxic cytokines and reactive oxygen and nitrite species is thought to contribute to the pathology of many neurodegenerative disorders. Agents that function to reduce microglial inflammatory activities are currently being sought. Work from our laboratory and others' has pointed to a role for P2 purinergic receptors (P2Rs) in reducing microglial production of inflammatory mediators [4,5,6]. The two major P2 receptor families are (page number not for citation purposes)
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