title: Phosphorylation-dependent subcellular localization of the small heat shock proteins HspB1/Hsp25 and HspB5/αB-crystallin in cultured hippocampal neurons.
abstract: The so-called stress response involving upregulation of heat shock proteins (Hsps) is a powerful mechanism of cells to deal with harmful conditions to which they are exposed throughout life, such as hyperthermia, hypoxia or oxidative stress. To gain more information about the molecular targets by which HspB1 (Hsp25) and HspB5 (αB-crystallin) might exert their neuroprotective effect we investigated the subcellular localization of unphosphorylated and phosphorylated HspB1 and B5 in neurons by immunocytochemistry and subcellular fractionation. In cultured hippocampal neurons, the unphosphorylated forms of both Hsps were localized in the perikaryon and nucleus, whereas the phosphorylated forms were recruited into neuronal processes. pHspB1-Ser15 and -Ser 86 were found within dendrites with a punctate distribution pattern partially colocalizing with the synaptic marker vGlut-1. pHspB5-Ser19 and -Ser45 localized to axons and dendrites with a filamentous-like staining pattern, whereas pHspB5-Ser59 was found in dendrites, especially along the plasma membrane and in spines. Biochemical analysis, i.e. subcellular fractionation of rat brain with subsequent Western blotting supported these localizations. These data show that in neurons HspB1 and B5 may have various molecular interaction partners at synapses, within dendrites and axons and that this interaction is likely to be regulated by phosphorylation. Stress-induced phosphorylation of HspB1 and B5 may lead to binding of these Hsps to their targets at synapses and neuronal processes which might provide one important mechanism of how they exert their neuroprotective effect.
authors.affiliation: Institute of Anatomy and Cell Biology, University of Ulm, Albert-Einstein-Allee 11, 89081, Ulm, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Schmidt
authors.forename: Britta
authors.initials: B
authors.lastname: Bartelt-Kirbach
authors.forename: Nikola
authors.initials: N
authors.lastname: Golenhofen
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D055551:HSP27 Heat-Shock Proteins / Q000032:analysis* / Q000378:metabolism; D050888:Heat-Shock Proteins, Small / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism*; D007150:Immunohistochemistry; D009474:Neurons / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D012694:Serine / Q000235:genetics / Q000378:metabolism; D038203:alpha-Crystallin B Chain / Q000032:analysis* / Q000378:metabolism
chemical_list: D055551:HSP27 Heat-Shock Proteins; D050888:Heat-Shock Proteins, Small; C486114:Hspb1 protein, rat; D038203:alpha-Crystallin B Chain; D012694:Serine