Integration of steroid hormone initiated membrane action to genomic function in the brain.
Estrogen is a ligand for the estrogen receptor (ER), which on binding 17beta-estradiol, functions as a ligand-activated transcription factor and regulates the transcription of target genes. This is the slow genomic mode of action. However, rapid non-genomic actions of estrogen also exist at the cell membrane. Using a novel two-pulse paradigm in which the first pulse rapidly initiates non-genomic actions using a membrane-limited estrogen conjugate (E-BSA), while the second pulse promotes genomic transcription from a consensus estrogen response element (ERE), we have demonstrated that rapid actions of estrogen potentiate the slower transcriptional response from an ERE-reporter in neuroblastoma cells. Since rapid actions of estrogen activate kinases, we used selective inhibitors in the two-pulse paradigm to determine the intracellular signaling cascades important in such potentiation. Inhibition of protein kinase A (PKA), PKC, mitogen activated protein kinase (MAPK) or phosphatidylinositol 3-OH kinase (PI-3K) in the first pulse decreases potentiation of transcription. Also, our data with both dominant negative and constitutive mutants of Galpha subunits show that Galpha(q) initiates the rapid signaling cascade at the membrane in SK-N-BE(2)C neuroblastoma cells. We discuss two models of multiple kinase activation at the membrane Pulses of estrogen induce lordosis behavior in female rats. Infusion of E-BSA into the ventromedial hypothalamus followed by 17beta-estradiol in the second pulse could induce lordosis behavior, demonstrating the applicability of this paradigm in vivo. A model where non-genomic actions of estrogen couple to genomic actions unites both aspects of hormone action.
Vasudevan
Nandini
N
Department of Biology, Pennsylvania State University, University Park, PA 16802, USA. nuv1@psu.edu
Kow
Lee Ming
LM
Pfaff
Donald
D
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol; D004967:Estrogens / Q000378:metabolism; D016678:Genome; D006728:Hormones / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008954:Models, Biological; D009447:Neuroblastoma / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D051381:Rats; D011960:Receptors, Estrogen / Q000378:metabolism; D015398:Signal Transduction; D013256:Steroids / Q000378:metabolism*; D013997:Time Factors; D014158:Transcription, Genetic
D004791:Enzyme Inhibitors; D004967:Estrogens; D006728:Hormones; D008024:Ligands; D011960:Receptors, Estrogen; D013256:Steroids; D004958:Estradiol