Transglutaminase (TG, EC 2.3.2.13) activity and levels of putrescine (a natural acyl-acceptor in the transglutaminase reaction) were measured in rat brains after 30 min ischemia and 8 or 24 h recirculation. TG activity was significantly increased in the striatum and hippocampus already during cerebral ischemia and, more pronounced, after 8 and 24 h recirculation. In the cortex, in contrast, TG activity did not change during ischemia and 8 h recirculation but was significantly increased after 24 h recirculation. Putrescine levels were sharply increased after 8 h recirculation and even further after 24 h recirculation. It is suggested that in vivo during ischemia and early recirculation, when cells are overloaded with calcium ions, a pathological increase in the TG-catalyzed cross-linking of proteins may be apparent especially in the nerve endings of the hippocampus where the intrinsic concentration of the acyl-donor (protein-bound glutamyl-moiety) has been shown to be high.