RESUMO
Adenylate cyclase (AC) has a specific sensitivity to Ca2+/calmodulin. AC-I, one of the mediator of learning and memory, plays an important role in signal transduction underlying learning and memory function. In the present study, we found ischemia-related changes of AC-I in the hippocampal CA1 region, but not in the CA2/3 region, after 5 min of transient forebrain ischemia in gerbils. In the sham-operated group, AC-I immunoreactive neurons were detected in pyramidal and non-pyramidal cells in the hippocampus proper. AC-I immunoreactivity was significantly increased at 3 h in the CA1 region after ischemic insult. Thereafter, AC-I immunoreactivity was gradually decreased. Four days after ischemic insult, AC-I-immunoreactive CA1 pyramidal cells in the stratum pyramidale were very few due to delayed neuronal death. The results of Western blot analysis showed that changes of AC-I protein contents were similar to immunohistochemical data after ischemic insult. Gpp(NH)p-dependent AC-I activity in hippocampal CA1 region was not changed in all groups, while Ca2+/calmodulin-dependent AC-I activity in hippocampal CA1 region was significantly decreased 24 h after ischemia-reperfusion. These results suggest that the decrease of AC-I activity may be associated with impairment of neurodevelopment and neuroplasticity including learning and memory although the AC-I immunoreactivity was maintained 24 h postischemic group compared to that of the sham-operated group.
Assuntos
Adenilil Ciclases/metabolismo , Hipocampo/enzimologia , Ataque Isquêmico Transitório/metabolismo , Ataque Isquêmico Transitório/patologia , Análise de Variância , Animais , Western Blotting/métodos , Modelos Animais de Doenças , Gerbillinae , Hipocampo/patologia , Imuno-Histoquímica/métodos , Masculino , Neurônios/metabolismo , Reperfusão , Coloração e Rotulagem/métodos , Fatores de TempoRESUMO
We observed chronological changes of platelet endothelial cell adhesion molecule-1 (PECAM-1), final mediator of neutrophil transendothelial migration, immunoreactivity, and protein level in the gerbil hippocampus proper after 5 min of transient ischemia. One day after ischemic insult, PECAM-1 immunoreactivity and protein level increased slightly in the hippocampus proper. Thereafter, PECAM-1 immunoreactivity and protein level increased significantly in the hippocampus proper by 4 days after ischemic insult. Especially, PECAM-1 in the hippocampal CA1 region was higher than that in the CA2/3 region. Five days after ischemic insult, PECAM-1 immunoreactivity decreased compared to the 4 days post-ischemic group. However, the RNA levels of PECAM-1 in the hippocampus proper were significantly decreased in the 4 days post-ischemic groups compared to that in the sham-operated group. This result suggests that the increase of PECAM-1 and decrease of PECAM-1 RNA in the CA1 region 4 days after ischemia may be associated with transmigration of neurotrophil.
Assuntos
Hipocampo/metabolismo , Isquemia/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Animais , Northern Blotting/métodos , Western Blotting/métodos , Contagem de Células/métodos , Gerbillinae , Hipocampo/patologia , Imuno-Histoquímica/métodos , Isquemia/patologia , Masculino , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , RNA Mensageiro/metabolismo , Reperfusão/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Fatores de TempoRESUMO
It is known that the hippocampus has vital functions in learning and memory, behavioral regulation, and activity-dependent synaptic plasticity, and that the hippocampus contains high levels of brain-derived neurotrophic factor (BDNF). In the present study, we followed age-dependent changes of BDNF immunoreactivity and protein level in the gerbil hippocampus to identify the correlation between BDNF and aging. BDNF immunoreactivity and its protein level significantly increased at postnatal month (PM) 12 in the hippocampus and thereafter reduced. At PM 24, BDNF immunoreactivity in the hippocampal CA1 region and dentate gyrus was similar to that in the PM 1 group, whereas BDNF immunoreactivity in the CA2/3 region at PM 24 was higher than that at PM 1. In the PM 24 group, an age-related neuronal loss and the decrease of reference and working memory were observed. In conclusion, our results suggest that observed reduction in BDNF and reference memory may be associated with age-dependent neuronal loss in the hippocampal CA1 region.
Assuntos
Envelhecimento/fisiologia , Fator Neurotrófico Derivado do Encéfalo/biossíntese , Hipocampo/metabolismo , Memória/fisiologia , Neurônios/metabolismo , Animais , Comportamento Animal/fisiologia , Contagem de Células , Giro Denteado/citologia , Giro Denteado/metabolismo , Gerbillinae , Hipocampo/citologia , Imuno-Histoquímica , Masculino , Aprendizagem em Labirinto/fisiologia , Neurônios/citologiaRESUMO
Immortalization-upregulated protein 1 (IMUP-1) and immortalization-upregulated protein 2 (IMUP-2) genes have been recently cloned and are known to be involved in SV40-mediated immortalization. IMUP-1 and IMUP-2 genes were strongly expressed in various cancer cell lines and tumors, suggesting the possibility that they might be involved in tumorigenicity. To directly elucidate the functional role of IMUP-1 and IMUP-2 on neoplastic transformation and tumorigenicity, we stably transfected IMUP-1 and IMUP-2 into NIH/3T3 mouse fibroblast cells. Cellular characteristics of the neoplastic transformation were assessed by transformation foci, growth in soft agar, and tumor development in nude mice. We found that IMUP-1 and IMUP-2 overexpressing cells showed altered growth properties, anchorage-independent growth in soft agar and inducing tumor in nude mice. Furthermore, IMUP-1 and IMUP-2 transformants proliferated in reduced serum and shortened cell cycle. These results suggest that ectopic overexpression of IMUP-1 and IMUP-2 may play an important role in acquiring a transformed phenotype, tumorigenicity in vivo, and be related to cellular proliferation.