title: Primary glia expressing the G93A-SOD1 mutation present a neuroinflammatory phenotype and provide a cellular system for studies of glial inflammation.
abstract: Detailed study of glial inflammation has been hindered by lack of cell culture systems that spontaneously demonstrate the "neuroinflammatory phenotype". Mice expressing a glycine --> alanine substitution in cytosolic Cu, Zn-superoxide dismutase (G93A-SOD1) associated with familial amyotrophic lateral sclerosis (ALS) demonstrate age-dependent neuroinflammation associated with broad-spectrum cytokine, eicosanoid and oxidant production. In order to more precisely study the cellular mechanisms underlying glial activation in the G93A-SOD1 mouse, primary astrocytes were cultured from 7 day mouse neonates. At this age, G93A-SOD1 mice demonstrated no in vivo hallmarks of neuroinflammation. Nonetheless astrocytes cultured from G93A-SOD1 (but not wild-type human SOD1-expressing) transgenic mouse pups demonstrated a significant elevation in either the basal or the tumor necrosis alpha (TNFalpha)-stimulated levels of proinflammatory eicosanoids prostaglandin E2 (PGE2) and leukotriene B4 (LTB4); inducible nitric oxide synthase (iNOS) and *NO (indexed by nitrite release into the culture medium); and protein carbonyl products. Specific cytokine- and TNFalpha death-receptor-associated components were similarly upregulated in cultured G93A-SOD1 cells as assessed by multiprobe ribonuclease protection assays (RPAs) for their mRNA transcripts. Thus, endogenous glial expression of G93A-SOD1 produces a metastable condition in which glia are more prone to enter an activated neuroinflammatory state associated with broad-spectrum increased production of paracrine-acting substances. These findings support a role for active glial involvement in ALS and may provide a useful cell culture tool for the study of glial inflammation.
authors.affiliation: Free Radical Biology and Aging Research Program, Oklahoma Medical Research Foundation (OMRF), 825 NE 13th Street, Oklahoma City, OK 73104, USA . kenneth-hensley@omrf.ouhsc.edu
authors.forename: Kenneth
authors.initials: K
authors.lastname: Hensley
authors.forename: Haitham
authors.initials: H
authors.lastname: Abdel-Moaty
authors.forename: Jerrod
authors.initials: J
authors.lastname: Hunter
authors.forename: Molina
authors.initials: M
authors.lastname: Mhatre
authors.forename: Shenyun
authors.initials: S
authors.lastname: Mou
authors.forename: Kim
authors.initials: K
authors.lastname: Nguyen
authors.forename: Tamara
authors.initials: T
authors.lastname: Potapova
authors.forename: Quentin N
authors.initials: QN
authors.lastname: Pye
authors.forename: Min
authors.initials: M
authors.lastname: Qi
authors.forename: Heather
authors.initials: H
authors.lastname: Rice
authors.forename: Charles
authors.initials: C
authors.lastname: Stewart
authors.forename: Katharine
authors.initials: K
authors.lastname: Stroukoff
authors.forename: Melinda
authors.initials: M
authors.lastname: West