Twelve PTEN's That Will Rock n roll This Current Year

Identical results were obtained with dissociated hippocampal neurons (Figure?7F). We have also tested whether glutamate can induce the decay of?miRNAs in ES-cell-derived neurons cultured for 15 days. The addition of glutamate, or NBQX/CPP plus glutamate, had no effect on the turnover of any miRNA (Figure?7G). To follow the electrophysiological properties of the ES-cell-derived neurons as a function of time in culture, we cultured them on microelectrode arrays (MEAs) and recorded their spontaneous spiking activity. Although some spiking was already visible after 10 to 15 days, maximal activity was reached after 25�C30 days of culturing (Figure?S7I). In summary, these experiments indicate that in both ES-cell-derived and hippocampal neurons neuronal activity may have both stimulatory and inhibitory PD0332991 mouse effects on miRNA turnover. Experiments with differentiated ES cells also suggest that neurons have to reach a certain level of maturity or connectivity to manifest activity-dependent changes in miRNA decay. In this work, we identified miRNAs which respond PTEN to different light conditions in mouse retinal neurons, independent of the circadian clock. Levels of the sensory neuron-specific miR-183/96/182 cluster, and miR-204 and -211, were downregulated during dark adaptation and upregulated during light adaptation, with rapid miRNA decay and increased transcription being responsible for the respective changes. We identified the voltage-dependent glutamate transporter, Slc1a1, as one of the targets of the light-regulated miR-183/96/182 cluster; other likely targets of these miRNAs in photoreceptor cells include Paip2b and Atp1b3. We found that many miRNAs in retinal, and also nonretinal, neurons turn over much faster than miRNAs in other cell types. Blocking action potentials with TTX, or glutamate receptors with NBQX/CPP, strongly affected miRNA turnover rates, indicating that active miRNA metabolism may be important for the function of neurons. Different algorithms were previously used to predict targets of miRNAs expressed in the retina, and, in a few instances, TSA HDAC clinical trial the validity of the predictions was tested using 3��-UTR reporter fusions in HEK293 or NIH 3T3 cells overexpressing retinal miRNA mimics (Arora et?al., 2007?and?Xu et?al., 2007). Using three different algorithms, we compiled a list of potential miR183/96/182 targets, including 214 mRNAs expressed in the mouse retina. Levels of 12 of them were up by more than 2-fold (p