Glucagon-like peptide-1 (GLP-1) and its analog exendin-4 (EX) have been considered as a growth factor implicated in pancreatic islet mass increase and -cell proliferation. This study aimed to investigate the effect of EX on cyclin D1 expression, a key regulator of the cell cycle, in the pancreatic -cell line INS-1. We demonstrated that EX significantly increased cyclin D1 mRNA and subsequently its protein levels. Although EX induced phosphorylation of Raf-1 and extracellular-signal-regulated kinase (ERK), both PD98059 and exogenous ERK1 had no effect on the cyclin D1 induction by EX. Instead, the cAMP-elevating agent forskolin induced cyclin D1 expression remarkably and this response was inhibited by pretreatment with H-89, a protein kinase A (PKA) inhibitor. Promoter analyses revealed that the cAMP-responsive element (CRE) site (at position -48; 5 -TAACGTCA-3 ) of cyclin D1 gene was required for both basal and EX-induced activation of the cyclin D1 promoter, which was confirmed by site-directed mutagenesis study. For EX to activate the cyclin D1 promoter effectively, CRE-binding protein (CREB) should be phosphorylated and bound to the putative CRE site, according to the results of electrophoretic mobility shift and chromatin immunoprecipitation assays. Lastly, a transfection assay employing constitutively active or dominant-negative CREB expression plasmids clearly demonstrated that CREB was largely involved in both basal and EX-induced cyclin D1 promoter activities. Taken together, EXinduced cyclin D1 expression is largely dependent on the cAMP/PKA signaling pathway, and EX increases the level of phosphorylated CREB and more potently transactivates cyclin D1 gene through binding of the CREB to the putative CRE site, implicating a potential mechanism underlying -cell proliferation by EX.

In this study we investigated the expression of brain-derived neurotrophic factor (BDNF) and c-fos mRNA in the hippocampal formation after febrile seizures (FSs) with in situ hybridization histochemistry using riboprobes. The induction of BDNF mRNA was firstly observed in the dentate gyrus at 30 min after FSs. The expression in the dentate gyrus peaked at 3 h and returned to basal level at 24 h. It was also observed in the CA3 of hippocampus from 2 to 3 h. The induction of c-fos mRNA was observed in the dentate gyrus at 30 min and 1 h. These observations suggest that BDNF and c-fos are the genes whose expression can be altered by FSs and might be related to pathologic alterations after FSs.

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