After Training For Selectively Target Extinction Processes ( Ref Aguilar Valles Et Al

After Training For Selectively Target Extinction Processes ( Ref Aguilar Valles Et Al

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after training to selectively target extinction processes (ref Aguilar-Valles et al., 2014). Puthanveettil labs use this approach as well as injection of specific viruses expressing shRNAs to study gene function in different brain regions. The appropriate siRNA will be infused into the CA1 of awake mice, at the time point indicated by our validation tests to produce significant lncRNA knockdown and we will begin CFC training. Fear memory will be tested 24 hrs later. Control animals will treated the same, with the exception of receiving an infusion of a non-targeting control siRNA (Aguilar-Valles, et al., 2014). We anticipate that siRNAs against lncRNAs that are critical for consolidation will produce decreased consolidation when compared to controls. If the contextual fear memory is significantly decreased, we will conclude that loss of the lncRNA prevented consolidation of the memory. If there is no effect of the siRNA at 24 hr test, we will conclude that the lncRNA is not necessary for consolidation.
C3.5b. Determine which of the validated lncRNAs from Aim 1 are capable of accelerating extinction. As indicated above, we will focus on elucidating the function of lncRNAs that are downregulated during extinction learning. The procedure here will be identical to 4a, the only difference being the expected outcome. Following establishing the efficacy of siRNAs/asRNAs to knockdown specific lncRNAs, we will micro-infuse them into Prl or Infrl mPFC of awake mice at the time points identified from our validation tests to produce significant lncRNA knockdown and will begin extinction training. Control animals will be treated the same with the exception of receiving an infusion of a non targeting control siRNA. We anticipate that siRNAs ag...


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...es (polyribosomes) by qRT-PCR. The presence of a specific lncRNA in polyribosomes would be a strong indicator of that lncRNA playing a role in translation (Flintoft., 2013; Guttman et al., 2013). We anticipate that these studies will begin to unravel the mechanism of action of specific lncRNAs and to identify their mRNA targets.
C3.7. Expected outcomes. We anticipate that studies described in this proposal will lead to the identification of specific lncRNAs that are differentially regulated in CA1 for fear memory consolidation and Prl and Infrl regions for extinction of fear memories. Furthermore, we will determine whether lncRNA can be used as tools to manipulate extinction contextual and cued fear conditioning models. Promising results will lead to the development of new strategies for therapeutically accelerating extinction, either through the lncRNA itself or

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