Time course of protein changes following in vitro ischemia in the rat hippocampal slice.
Following 5 min in vitro ischemia, total protein synthesis is dramatically and persistently inhibited in neurons in the rat hippocampal slice. This model system was used to explore the responses of individual proteins to this irreversible insult. In vitro ischemia inhibited new protein synthesis of most proteins analyzed; however, the synthesis of a 68/70 kDa protein was substantially stimulated for the first hour after ischemia. By 3 hr postischemia, its synthesis rates were depressed to 60% of control rates. Although the total amounts of most proteins were not significantly depleted for the first few hours after ail ischemic episode, there were several notable exceptions. The levels of HSC73, a constitutively expressed member of the 70 kDa stress protein family, were reduced after in vitro ischemia. In addition, MAP-2 (microtubule-associated protein-2) and alpha-tubulin were depleted in the early hours after the insult, with MAP-2 exhibiting a detectable depletion earlier than tubulin. In contrast, the levels and distribution of a 68 kDa neurofilament protein localized to CA3 pyramidal neurons in the slice, apparently distinct from the band whose new synthesis was stimulated, were not affected by the 5 min in vitro ischemia insult. Thus, the responses of individual proteins to ischemia varied considerably, These individual responses could play an important role in the damage mechanism that is initiated in response to in vitro ischemia.
Raley-Susman
K M
KM
Vassar College, Department of Biology, Poughkeepsie, NY 12601, USA.
Murata
J
J
D000818:Animals; D002545:Brain Ischemia / Q000378:metabolism* / Q000473:pathology; D004396:Coloring Agents; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003720:Densitometry; D006360:Heat-Shock Proteins / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism* / Q000473:pathology; D066298:In Vitro Techniques; D008297:Male; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012834:Silver; D013997:Time Factors
D004396:Coloring Agents; D003598:Cytoskeletal Proteins; D006360:Heat-Shock Proteins; D009419:Nerve Tissue Proteins; D012834:Silver