C2A. Preliminary Results: C2A1. Establishment of fear conditioning protocols to study consolidation and extinction of fear memories: Presented below is published and preliminary data to establish feasibility for the proposed studies; the conditioning and extinction protocols are sensitive to manipulations that enhance or impair fear memory storage and the acquisition and consolidation of extinction. We reported (Zhang et al. 2013) that mice given post-training serotonin 2A receptor agonist TCB-2 (1mg/kg, open bar) exhibited significantly greater freezing than vehicle-treated mice (black bar) during a context test 24h after fear conditioning (A). These data indicate that activation of 5HT2A receptors enhances storage of fear memory. In a second
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RESEARCH DESIGN AND METHODOLOGY: As described in the preliminary data, all of the behavioral, genomics, bioinformatics, gene manipulation and biochemical studies are feasible between the Puthanveettil and Stackman laboratories. Additionally, we have obtained support letters from the leading experts in the field of lncRNA biology and regulation, such as John Rinn from Harvard University and the Broad Institute and our colleague at Scripps, Kevin Morris. Letters for genomics, bioinformatics, mouse behavior and imaging analysis are also attached. Below we briefly describe experimental design and methodologies to achieve the two …show more content…
Identification of spatial and temporal changes in lncRNA expression following consolidation or extinction. To identify specific lncRNAs whose levels change significantly in the CA1/Prl/Ifrl, RNAseq analysis will be performed on total RNAs isolated from these brain regions following consolidation or extinction training. Because several studies (Rudenko et al., 2013; Wang et al., 2012; Merlo et al., 2008; Mamiya et al., 2009; Zhang et al., 2006; von Hertzen and Giese., 2005; Vianna et al., 2005; Mac Callum et al., 2014; Huynh et al., 2014; Blundell et al., 2008; Morris et al., 2006; Debiec et al., 2002) have shown that changes in transcription and translation immediately following memory retrieval are critical for consolidation and extinction learning, we will study changes in lncRNAs 30 minutes, 1 hour and 6 hours after training (Aim 1 and 2). Specific changes in lncRNAs are anticipated to have long-lasting effects on transcription and translation because of the known mechanisms of lncRNA regulation of gene expression. Several studies have suggested that products of experience-induced changes in transcription will be used later for the structural changes associated with long-term memory storage (Kandel et al., 2014; Bailey et al., 2004). Hence, data on early changes in lncRNAs
RESEARCH DESIGN AND METHODOLOGY: As described in the preliminary data, all of the behavioral, genomics, bioinformatics, gene manipulation and biochemical studies are feasible between the Puthanveettil and Stackman laboratories. Additionally, we have obtained support letters from the leading experts in the field of lncRNA biology and regulation, such as John Rinn from Harvard University and the Broad Institute and our colleague at Scripps, Kevin Morris. Letters for genomics, bioinformatics, mouse behavior and imaging analysis are also attached. Below we briefly describe experimental design and methodologies to achieve the two …show more content…
Identification of spatial and temporal changes in lncRNA expression following consolidation or extinction. To identify specific lncRNAs whose levels change significantly in the CA1/Prl/Ifrl, RNAseq analysis will be performed on total RNAs isolated from these brain regions following consolidation or extinction training. Because several studies (Rudenko et al., 2013; Wang et al., 2012; Merlo et al., 2008; Mamiya et al., 2009; Zhang et al., 2006; von Hertzen and Giese., 2005; Vianna et al., 2005; Mac Callum et al., 2014; Huynh et al., 2014; Blundell et al., 2008; Morris et al., 2006; Debiec et al., 2002) have shown that changes in transcription and translation immediately following memory retrieval are critical for consolidation and extinction learning, we will study changes in lncRNAs 30 minutes, 1 hour and 6 hours after training (Aim 1 and 2). Specific changes in lncRNAs are anticipated to have long-lasting effects on transcription and translation because of the known mechanisms of lncRNA regulation of gene expression. Several studies have suggested that products of experience-induced changes in transcription will be used later for the structural changes associated with long-term memory storage (Kandel et al., 2014; Bailey et al., 2004). Hence, data on early changes in lncRNAs