title: Rapid manifestation of reactive astrogliosis in acute hippocampal brain slices.
abstract: A flurry of studies over the past decade has shown that astrocytes play a more active role in neural function than previously recognized. Hippocampal slices prepared from young rodent pups have served as a popular model for studying the pathways by which astrocytes participate in synaptic transmission. It is, however, not known how well astrocytes tolerate traumatic injury and hypoxia, which are unavoidable when preparing acute slices. We here showed that astrocytes exhibit striking changes in expression of several receptors and structural proteins, including re-expression of the developmental marker nestin within 90 min following preparation of live vibratome slices. Moreover, immunoelectron microscopy showed a 2.7-fold loss of astrocytic processes in acute hippocampal slices prepared from glial fibrillary acidic protein-green fluorescent protein reporter mice. A sharp decrease in the number of mitochondria was also noted in acute slices, concurrently with an increase in mitochondrial size. Glycogen content decreased 3-fold upon slice preparation and did not recover despite stable recordings of field excitatory postsynaptic current. Analysis of Ca(2+) signaling showed that astrocytic responses to purine receptor and mGluR5 agonists differed in slice versus in vivo. These observations suggest that the functional properties and the fine structure of astrocytes in slices may be reflective of early stages of reactive gliosis and should be confirmed in vivo when possible.
authors.affiliation: Division of Glial Disease and Therapeutics, Center for Translational Neuromedicine, University of Rochester Medical Center, Rochester, New York.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Takano
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Han
authors.forename: Fushun
authors.initials: F
authors.lastname: Wang
authors.forename: Qiwu
authors.initials: Q
authors.lastname: Xu
authors.forename: Xiaohai
authors.initials: X
authors.lastname: Wang
authors.forename: Nancy Ann
authors.initials: NA
authors.lastname: Oberheim Bush
authors.forename: Nancy
authors.initials: N
authors.lastname: Cruz
authors.forename: Gerald A
authors.initials: GA
authors.lastname: Dienel
authors.forename: Maiken
authors.initials: M
authors.lastname: Nedergaard
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D051401:Aquaporin 4 / Q000378:metabolism; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005911:Gliosis / Q000209:etiology / Q000473:pathology*; D006003:Glycogen / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology* / Q000293:injuries*; D066298:In Vitro Techniques; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D051379:Mice; D009243:NAD / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology
chemical_list: D051401:Aquaporin 4; D005904:Glial Fibrillary Acidic Protein; D000644:Quaternary Ammonium Compounds; D009243:NAD; D049452:Green Fluorescent Proteins; D019344:Lactic Acid; D006003:Glycogen; C027917:tetramethylammonium; D002118:Calcium
keywords: NADH; astrohcytes; electron microscopy; interstitial space; mitochondria; purinergic receptors; two-photon microscopy