title: Deimination level and peptidyl arginine deiminase 2 expression are elevated in astrocytes with increased incubation temperature.
abstract: Astrocytes respond to environmental cues, including changes in temperatures. Increased deimination, observed in many progressive neurological diseases, is thought to be contributed by astrocytes. We determined the level of deimination and expression of peptidyl arginine deiminase 2 (PAD2) in isolated primary astrocytes in response to changes on either side (31°C and 41°C) of the optimal temperature (37°C). We investigated changes in the astrocytes by using a number of established markers and accounted for cell death with the CellTiter-Blue assay. We found increased expression of glial fibrillary acidic protein, ALDH1L1, and J1-31, resulting from increased incubation temperature and increased expression of TSP1, S100β, and AQP4, resulting from decreased incubation temperature vs. optimal temperature, suggesting activation of different biochemical pathways in astrocytes associated with different incubation temperatures. Mass spectrometric analyses support such trends. The PAD2 level was increased only as a result of increased incubation temperature with a commensurate increased level of deimination. Actin cytoskeleton and iso[4]LGE, a lipid peroxidase modification, also showed an increase with higher incubation temperature. Altogether, these results suggest that temperature, as an environmental cue, activates astrocytes in a different manner on either side of the optimal temperature and that increase in deimination is associated only with the higher temperature side of the spectrum.
authors: Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami, Miami, Florida.|CIBICI, Department of Clinical Biochemistry, Faculty of Chemistry, National University of Córdoba, Córdoba, República Argentina.
authors: Mabel
authors: M
authors: Enriquez-Algeciras
authors: Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami, Miami, Florida.
authors: Sanjoy K
authors: SK
authors: Bhattacharya
authors: CIBICI, Department of Clinical Biochemistry, Faculty of Chemistry, National University of Córdoba, Córdoba, República Argentina.
authors: Horacio M
authors: HM
authors: Serra
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D051401:Aquaporin 4 / Q000378:metabolism; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D001703:Biophysics; D003994:Bucladesine / Q000494:pharmacology; D002470:Cell Survival; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004797:Enzyme-Linked Immunosorbent Assay; D005904:Glial Fibrillary Acidic Protein; D006867:Hydrolases / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D000080064:Protein-Arginine Deiminase Type 2; D000076342:Protein-Arginine Deiminases; D064568:S100 Calcium Binding Protein beta Subunit / Q000378:metabolism; D013696:Temperature
chemical_list: C496031:Aqp4 protein, mouse; D051401:Aquaporin 4; D005904:Glial Fibrillary Acidic Protein; D064568:S100 Calcium Binding Protein beta Subunit; D003994:Bucladesine; D000444:Aldehyde Dehydrogenase; D006867:Hydrolases; C000620353:Padi2 protein, mouse; D000080064:Protein-Arginine Deiminase Type 2; D000076342:Protein-Arginine Deiminases
keywords: activation markers; astrocytes; deimination; environmental cues; peptidyl arginine deiminase; temperature incubation