Dynamic visualization of local protein synthesis in hippocampal neurons.
Using pharmacological approaches, several recent studies suggest that local protein synthesis is required for synaptic plasticity. Convincing demonstrations of bona fide dendritic protein synthesis in mammalian neurons are rare, however. We developed a protein synthesis reporter in which the coding sequence of green fluorescent protein is flanked by the 5' and 3' untranslated regions from CAMKII-alpha, conferring both dendritic mRNA localization and translational regulation. In cultured hippocampal neurons, we show that BDNF, a growth factor involved in synaptic plasticity, stimulates protein synthesis of the reporter in intact, mechanically, or "optically" isolated dendrites. The stimulation of protein synthesis is blocked by anisomycin and not observed in untreated neurons. In addition, dendrites appear to possess translational hot spots, regions near synapses where protein synthesis consistently occurs over time.
Aakalu
G
G
Caltech, Howard Hughes Medical Institute, Division of Biology 216-76, Pasadena, CA 91125, USA.
Smith
W B
WB
Nguyen
N
N
Jiang
C
C
Schuman
E M
EM
D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D000841:Anisomycin / Q000494:pharmacology; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics*; D002478:Cells, Cultured; D003712:Dendrites / Q000502:physiology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology / Q000502:physiology*; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D014176:Protein Biosynthesis / Q000187:drug effects / Q000502:physiology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology; D014162:Transfection
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D019208:Brain-Derived Neurotrophic Factor; D008164:Luminescent Proteins; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D000841:Anisomycin; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases