Profiling of transcription-factor activity in rat hippocampus

C.A. Caamao; N. Calvo; I. Morano; S. Watson; H. Akil
Society for Neuroscience annual meeting. 2005.


This work represents a first step towards the selection of actively regulated transcription factors (TFs) in hippocampus for their assessment in behavioral conditions, including learning and memory. The biochemical activity of different TFs provides a key to understanding how gene expression is regulated. In order to evaluate TF activity, multiplexed functional assays are required to supplement those measuring mRNA or protein levels. In this study based on isolation of protein-DNA complexes and DNA arrays, we measured TF activity by incubating nuclear extracts from hippocampal homogenates with a panel of 150 TF-response elements simultaneously under basal, adrenalectomy and corticosterone replacement conditions. Our arrays consistently detected a set of putative TFs, many of them in agreement with previous mRNA expression studies, and in several cases their DNA-binding activities were significantly affected by the changes in glucocorticoid levels. Selected TFs are being assessed by electrphoretic mobility-shift and mass spectrometry assays as putative downstream effectors of signaling proteins that we found to be differentially phosphorylated upon learning. Support Contributed By: NIH PO1 MH42251, RO1 DA13386, and N00014-02-1-0879. Also, CC, SW and HA are members of the Pritzker Neuropsychiatric Disorders Research Consortium which is supported by the Pritzker Neuropsychiatric Disorders Research Fund L.L.C.