A high-efficiency protein transduction system demonstrating the role of PKA in long-lasting long-term potentiation.
Proteins and peptides have been demonstrated to penetrate across the plasma membrane of eukaryotic cells by protein transduction domains. We show that protein transduction by 11 arginine (11R) is an efficient method of delivering proteins into the neurons of brain slices. Here, we demonstrate that PKA inhibitory peptide, fused with 11R and nuclear localization signal, delivers the peptide exclusively into the nuclear compartment of neurons in brain slices. This inhibitory peptide blocked both cAMP responsive element-binding protein phosphorylation and long-lasting long-term potentiation (LTP) induction, but not early LTP. These results highlight transduction of proteins and peptides into specific neuronal subcellular compartments in brain slices as a powerful tool for studying neuronal plasticity.
Matsushita
M
M
First Department of Physiology, Okayama University Medical School, Okayama 700-8558, Japan.
Tomizawa
K
K
Moriwaki
A
A
Li
S T
ST
Terada
H
H
Matsui
H
H
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D002451:Cell Compartmentation; D002454:Cell Differentiation / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects; D017930:Genes, Reporter; D006624:Hippocampus; D066298:In Vitro Techniques; D047908:Intracellular Signaling Peptides and Proteins; D017774:Long-Term Potentiation / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000235:genetics; D010455:Peptides / Q000235:genetics; D010766:Phosphorylation / Q000187:drug effects; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
D002352:Carrier Proteins; D017362:Cyclic AMP Response Element-Binding Protein; D047908:Intracellular Signaling Peptides and Proteins; D019913:Nuclear Localization Signals; D010455:Peptides; D011993:Recombinant Fusion Proteins; C015482:protein kinase modulator; C015462:polyarginine; D017868:Cyclic AMP-Dependent Protein Kinases