title: Distinct activation profiles in microglia of different ages: a systematic study in isolated embryonic to aged microglial cultures.
abstract: Microglia have been implicated in disease progression for several age-related brain disorders. However, while microglia's contribution to the progression of these disorders is accepted, the effect of aging on their endogenous cellular characteristics has received limited attention. In fact, a comprehensive study of how the structure and function of microglia changes as a function of developmental age has yet to be performed. Here, we describe the functional response characteristics of primary microglial cultures prepared from embryonic, neonatal (Neo), 2-3month-old, 6-8month-old, 9-11month-old, and 13-15month-old rats. Microglial morphology, glutamate (GLU) uptake, and release of trophic and inflammatory factors were assessed under basal conditions and in microglia activated with adenosine 5'-triphosphate (ATP) or lipopolysaccharide. We found that microglia from different age groups were both morphologically and functionally distinct. Upon activation by ATP, Neo microglia were the most reactive, upregulating nitric oxide, tumor necrosis factor-α, and brain-derived neurotrophic factor release as well as GLU uptake. This upregulation translated into neurotoxicity in microglia-neuron co-cultures that were not observed with microglia of different developmental ages. Interestingly, 13-15month-old microglia exhibited similar activation profiles to Neo microglia, whereas microglia from younger adults and embryos were activated less by ATP. Our data also identify age-dependent differences in purinergic receptor subtype expression that contribute to the regulation of neuronal survival. Combined, our data demonstrate that microglial activation and purinergic receptor profiles vary non-linearly with developmental age, a potentially important finding for studies examining the role of microglia in neurodegenerative disorders.
authors.affiliation: Neurochemical Research Unit, Department of Psychiatry and Centre for Neuroscience, University of Alberta, Edmonton, Alberta T6G 2K7, Canada.
authors.forename: A Y
authors.initials: AY
authors.lastname: Lai
authors.forename: C D
authors.initials: CD
authors.lastname: Dibal
authors.forename: G A
authors.initials: GA
authors.lastname: Armitage
authors.forename: I R
authors.initials: IR
authors.lastname: Winship
authors.forename: K G
authors.initials: KG
authors.lastname: Todd
mesh_terms: D000367:Age Factors; D000375:Aging / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000166:cytology / Q000196:embryology* / Q000378:metabolism*; D002478:Cells, Cultured; D018920:Coculture Techniques; D017628:Microglia / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
keywords: AG; ANOVA; ATP; BBG; BDNF; Brilliant Blue G; CM; DIV; DMEM; Dulbecco’s Modified Eagle Medium; EDTA; ELISA; Em; FBS; GFAP; GLU; Hyp; Iba1; LPS; MAP-2; NO; Neo; PBS; RB2; Reactive Blue 2; TNF-α; adenosine 5′-triphosphate; ageing; aminoguanidine; analysis of variance; brain-derived neurotrophic factor; conditioned media; cytokines; days in vitro; development; embryonic; enzyme-linked immunosorbent assays; ethylenediaminetetraacetic acid; fetal bovine serum; glial fibrillary acidic protein; glutamate; hypoxia; iNOS; inducible NO synthase; inflammation; ionized calcium-binding adaptor protein 1; lipopolysaccharide; microtubule-associated protein-2; neonatal; neurotoxicity; nitric oxide; phosphate-buffered saline; purinergic receptors; tumor necrosis factor-α