ARF6 bi-directionally regulates the dendritic spine formation depending on neuronal maturation and activity
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ABSTRACT: Recent studies reported contradictory results regarding the role of ADP-ribosylation factor 6 (ARF6), a small GTPase known to regulate actin cytoskeleton, in dendritic spine development and maintenance. We readdress this question, and found that ARF6 either positively or negatively regulates dendritic spine formation depending on neuronal maturation and activity. ARF6 activation facilitates filopodia to spines transition, increasing the spine formation in developing neurons while it decreases spine density in matured neurons. Consistently, genome-wide microarray analysis revealed that Arf6 activation in developing and matured neurons leads to opposite expression patterns of a subset of genes that are involved in neuronal morphology. Overall design: To readdress effects of ARF6-T157A on spine formation in both developing and matured neurons, we have generated the Sindbis virus for ARF6-T157A-6xHis and performed its infection into rat hippocampal neuron culture at 10 DIV (days in vitro) for developing stage and 17 DIV for mature stage, and maintained for additional 48 h. Infected neurons were washed twice with PBS and were frozen at -70C for storage. Samples were lysed and RNA isolation was carried out using the RNA-spin total RNA extraction kit (iNtRON Biotechnology, Sungnam, Kyungki-Do, South Korea). Then, genome-wide microarray analysis was performed. Microarray analysis revealed that Arf6 activation in developing and matured neurons leads to opposite expression patterns of a subset of genes that are involved in neuronal morphology.
INSTRUMENT(S): Agilent-028279 SurePrint G3 Rat GE 8x60K Microarray (Feature Number version)
SUBMITTER:
Minhyung Kim
PROVIDER: GSE40937 | GEO | 2015-04-02
SECONDARY ACCESSION(S): PRJNA175398
REPOSITORIES: GEO
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