Changes in heat shock protein 27 phosphorylation and immunocontent in response to preconditioning to oxygen and glucose deprivation in organotypic hippocampal cultures.
Organotypic hippocampal cultures have been recently used to study in vitro ischaemic neuronal death. Sub-lethal periods of ischaemia in vivo confer resistance to lethal insults and many studies have demonstrated the involvement of heat shock proteins in this phenomenon. We used organotypic hippocampal cultures to investigate the involvement of heat shock protein (HSP) 27 in preconditioning to oxygen and glucose deprivation. Neuronal damage was assessed using propidium iodide fluorescence; HSP27 phosphorylation and immunocontent were obtained using (32)Pi labelling followed by sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting. We observed that immunocontent of HSP27 was increased after lethal or sub-lethal treatment, indicating it is a response to metabolic stress. Treatments with 5 or 10 min of oxygen and glucose deprivation (OGD) or 1- microM N-methyl-D-aspartate (NMDA) induced tolerance to 40 min of OGD associated with an increase in HSP27 immunocontent and phosphorylation. These data suggest that, in vitro, phosphorylated HSP27 might be involved in preconditioning, probably acting as a modulator of actin filaments or by the blockage of neurodegenerative processes.
Valentim
L M
LM
Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, UFRGS, Porto Alegre 90035-003, Brazil.
Rodnight
R
R
Geyer
A B
AB
Horn
A P
AP
Tavares
A
A
Cimarosti
H
H
Netto
C A
CA
Salbego
C G
CG
D000818:Animals; D015153:Blotting, Western; D016923:Cell Death / Q000187:drug effects; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D005947:Glucose / Q000172:deficiency / Q000378:metabolism*; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006624:Hippocampus / Q000378:metabolism*; D000860:Hypoxia / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D019194:Ischemic Preconditioning / Q000379:methods; D016202:N-Methylaspartate / Q000494:pharmacology; D009924:Organ Culture Techniques; D010100:Oxygen / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D018690:Excitatory Amino Acid Agonists; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D016202:N-Methylaspartate; D005947:Glucose; D010100:Oxygen