title: Neuritic outgrowth associated with astroglial phenotypic changes induced by antisense glial fibrillary acidic protein (GFAP) mRNA in injured neuron-astrocyte cocultures.
abstract: In the adult CNS, axons fail to regenerate after injury. Among the cell interactions that lead to this failure are those developed with astrocytes. In an effort to elucidate the mechanisms underlying these negative interactions, we have used astrocytes treated with antisense glial fibrillary acidic protein (GFAP) mRNA to inhibit the formation of gliofilaments, indispensable for the astroglial morphological response to injury, and have studied their permissivity for neuritic outgrowth. In a neuron-astrocyte coculture, a mechanical lesion led to hypertrophy of astrocytes neighboring the lesion. Neuronal cell bodies and neurites were absent both from the area of lesion and from its surroundings. Reactive astrocytes appeared, therefore, to be a nonpermissive substrate. Transfection that used antisense GFAP mRNA blocked astroglial morphological changes and was characterized by both a persistence of neuronal cell bodies in the vicinity of the lesion site and a growth of neurites into the same region. These morphological differences were associated with a 46% decrease in the GFAP translation capacity and a 50% increase in the concentration of GAP-43 in the treated cultures. Neurons were associated mainly with an extracellular laminin network, which was predominant at the lesion site in treated cocultures. In contrast, those astrocytes highly laminin-immunoreactive appeared to be a nonpermissive substrate for neurons. These results show that inhibition in GFAP synthesis, leading to a reduction of astroglial hypertrophy, relieves the blockade of neuritic outgrowth that normally is observed after a lesion. The mechanisms may involve changes in the secretion of extracellular matrix molecules by astrocytes.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U421, IM3, Faculté de Médecine, 94010 Créteil, France.
authors.forename: T
authors.initials: T
authors.lastname: Lefrançois
authors.forename: C
authors.initials: C
authors.lastname: Fages
authors.forename: M
authors.initials: M
authors.lastname: Peschanski
authors.forename: M
authors.initials: M
authors.lastname: Tardy
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000737:chemistry* / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured / Q000737:chemistry / Q000502:physiology; D005455:Fluorescent Antibody Technique; D019922:GAP-43 Protein; D015870:Gene Expression / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis / Q000235:genetics*; D005911:Gliosis / Q000503:physiopathology; D007797:Laminin / Q000032:analysis; D008562:Membrane Glycoproteins / Q000032:analysis; D051379:Mice; D009419:Nerve Tissue Proteins / Q000032:analysis; D016501:Neurites / Q000737:chemistry / Q000502:physiology; D016900:Neurofilament Proteins / Q000032:analysis; D009474:Neurons / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D010641:Phenotype; D016372:RNA, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D014162:Transfection
chemical_list: D019922:GAP-43 Protein; D005904:Glial Fibrillary Acidic Protein; D007797:Laminin; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger