Integrated Analysis of miRNA/mRNA Expression in the Neurocircuitry Underlying Nicotine Dependence

Nicotine dependence is responsible for perpetuating the adverse health effects due to tobacco use, the leading cause of preventable death worldwide. Nicotine is an agonist for nicotinic acetylcholine receptors, which are enriched in the mesocorticolimbic and habenulo-interpeduncular circuitries, underlying nicotine reward and withdrawal, respectively. Drugs of abuse, including nicotine, induce stable neuroadaptations, requiring protein synthesis through regulation of transcription factors, epigenetic mechanisms, and non-coding RNAs. It also been shown that miRNAs in brain are regulated by nicotine and that miRNA dysregulation contributes to brain dysfunction, including drug addiction. While much is known about the neurocircuitry responsible for the behaviors associated with nicotine reward or withdrawal, the underlying molecular mechanisms of how these changes in behavior are induced are less clear. Using miRNA-/mRNA-Seq, we demonstrate that there are widespread changes in both miRNA and mRNA expression in brain regions comprising the mesocorticolimbic circuit after chronic nicotine treatment, and the habenulo-interpeduncular circuit during acute nicotine withdrawal. Conserved, differentially expressed miRNAs were predicted to target inversely regulated mRNAs. We determined that expression of miR-106b-5p is up-regulated and Profilin 2 (Pfn2), an actin-binding protein enriched in the brain, is down-regulated in the interpeduncular nucleus (IPN) during acute nicotine withdrawal. Further we show that miR-106b-5p represses Pfn2 expression. We demonstrate that knockdown of Pfn2 in the IPN is sufficient to induce anxiety, a symptom of withdrawal. This novel role of Pfn2 in nicotine withdrawal-associated anxiety is a prime example of this dataset’s utility, allowing for the identification of a multitude of miRNAs/mRNA which may participate in the molecular mechanisms underlying the neuroadaptations of nicotine dependence

National Survey of Sea Lice (Lepeophtheirus salmonis Krøyer and Caligus elongatus Nordmann) on Fish Farms in Ireland – 2019

Farmed stocks of Atlantic salmon in Ireland are inspected on 14 occasions throughout the year to monitor sea lice levels as part of a national programme. Sea lice are a naturally occurring parasite found on marine fish, including salmonids. They are small ecto-parasitic copepod crustaceans and there are approximately 559 species. The objectives of the National Sea Lice Monitoring Programme are: To provide an objective measurement of infestation levels on farms. To investigate the nature of infestations. To provide management information to drive the implementation of control and management strategies. To facilitate further development and refinement of this strategy. The sea lice control and management strategy has five principal components: Separation of generations. Annual fallowing of sites. Early harvest of two-sea-winter fish. Targeted treatment regimes, including synchronous treatments. Agreed husbandry practices

Integrated miRNA-/mRNA-Seq of the Habenulo-Interpeduncular Circuit During Acute Nicotine Withdrawal

Tobacco use is the leading preventable cause of mortality in the world. The limited number of smoking cessation aids currently available are minimally effective, highlighting the need for novel therapeutic interventions. We describe a genome-wide approach to identify potential candidates for such interventions. Next-generation sequencing was performed using RNA isolated from the habenulo-interpeduncular circuit of male mice withdrawn from chronic nicotine treatment. This circuit plays a central role in the nicotine withdrawal response. Differentially expressed miRNAs and mRNAs were validated using RT-qPCR. Many of the differentially expressed mRNAs are predicted targets of reciprocally expressed miRNAs. We illustrate the utility of the dataset by demonstrating that knockdown in the interpeduncular nucleus of a differentially expressed mRNA, that encoding profilin 2, is sufficient to induce anxiety-related behavior. Importantly, profilin 2 knockdown in the ventral tegmental area did not affect anxiety behavior. Our data reveal wide-spread changes in gene expression within the habenulo-interpeduncular circuit during nicotine withdrawal. This dataset should prove to be a valuable resource leading to the identification of substrates for the design of innovative smoking cessation aids

miRNAome analysis of the mammalian neuronal nicotinic acetylcholine receptor gene family

Nicotine binds to and activates a family of ligand-gated ion channels, neuronal nicotinic acetylcholine receptors (nAChRs). Chronic nicotine exposure alters the expression of various nAChR subtypes, which likely contributes to nicotine dependence; however, the underlying mechanisms regulating these changes remain unclear. A growing body of evidence indicates that microRNAs (miRNAs) may be involved in nAChR regulation. Using bioinformatics, miRNA library screening, site-directed mutagenesis, and gene expression analysis, we have identified a limited number of miRNAs that functionally interact with the 3\u27-untranslated regions (3\u27 UTRs) of mammalian neuronal nAChR subunit genes. In silico analyses revealed specific, evolutionarily conserved sites within the 3\u27 UTRs through which the miRNAs regulate gene expression. Mutating these sites disrupted miRNA regulation confirming the in silico predictions. In addition, the miRNAs that target nAChR 3\u27 UTRs are expressed in mouse brain and are regulated by chronic nicotine exposure. Furthermore, we show that expression of one of these miRNAs, miR-542-3p, is modulated by nicotine within the mesocorticolimbic reward pathway. Importantly, overexpression of miR-542-3p led to a decrease in the protein levels of its target, the nAChR beta2 subunit. Bioinformatic analysis suggests that a number of the miRNAs play a general role in regulating cholinergic signaling. Our results provide evidence for a novel mode of nicotine-mediated regulation of the mammalian nAChR gene family

miRNAome analysis of the mammalian neuronal nicotinic acetylcholine receptor gene family

Nicotine binds to and activates a family of ligand-gated ion channels, neuronal nicotinic acetylcholine receptors (nAChRs). Chronic nicotine exposure alters the expression of various nAChR subtypes, which likely contributes to nicotine dependence; however, the underlying mechanisms regulating these changes remain unclear. A growing body of evidence indicates that microRNAs (miRNAs) may be involved in nAChR regulation. Using bioinformatics, miRNA library screening, site-directed mutagenesis, and gene expression analysis, we have identified a limited number of miRNAs that functionally interact with the 3\u27-untranslated regions (3\u27 UTRs) of mammalian neuronal nAChR subunit genes. In silico analyses revealed specific, evolutionarily conserved sites within the 3\u27 UTRs through which the miRNAs regulate gene expression. Mutating these sites disrupted miRNA regulation confirming the in silico predictions. In addition, the miRNAs that target nAChR 3\u27 UTRs are expressed in mouse brain and are regulated by chronic nicotine exposure. Furthermore, we show that expression of one of these miRNAs, miR-542-3p, is modulated by nicotine within the mesocorticolimbic reward pathway. Importantly, overexpression of miR-542-3p led to a decrease in the protein levels of its target, the nAChR beta2 subunit. Bioinformatic analysis suggests that a number of the miRNAs play a general role in regulating cholinergic signaling. Our results provide evidence for a novel mode of nicotine-mediated regulation of the mammalian nAChR gene family

National survey of sea lice (Lepeophtheirus salmonis Kroyer and Caligus elongatus Nordmann) on fish farms in Ireland - 2016

Farmed stocks of Atlantic salmon in Ireland are inspected on 14 occasions throughout the year to monitor sea lice levels as part of a national programme

National Survey Of Sea Lice (Lepeophtheirus salmonis Krøyer and Caligus elongatus Nordmann) on Fish Farms in Ireland – 2022

Farmed stocks of Atlantic salmon in Ireland are inspected on 14 occasions throughout the year to monitor sea lice levels as part of a national programme. Sea lice are a naturally occurring parasite found on marine fish, including salmonids. They are small ecto-parasitic copepod crustaceans and there are approximately 559 species. The objectives of the National Sea Lice Monitoring Programme are: *To provide an objective measurement of infestation levels on farms. * To investigate the nature of infestations. * To provide management information to drive the implementation of control and management strategies. * To facilitate further development and refinement of this strategy. The sea lice control and management strategy has five principal components: * Separation of generations. * Annual fallowing of sites. * Early harvest of two-sea-winter fish. * Targeted treatment regimes, including synchronous treatments. * Agreed husbandry practices.Marine Institut

National Survey of Sea Lice (Lepeophtheirus salmonis Krøyer and Caligus elongatus Nordmann) on Fish Farms in Ireland – 2018

Farmed stocks of Atlantic salmon in Ireland are inspected on 14 occasions throughout the year to monitor sea lice levels as part of a national programme. Sea lice are a naturally occurring parasite found on marine fish, including salmonids. They are small ecto-parasitic copepod crustaceans and there are approximately 559 species. The objectives of the National Sea Lice Monitoring Programme are: To provide an objective measurement of infestation levels on farms. To investigate the nature of infestations. To provide management information to drive the implementation of control and management strategies. To facilitate further development and refinement of this strategy. The sea lice control and management strategy has five principal components: Separation of generations. Annual fallowing of sites. Early harvest of two-sea-winter fish. Targeted treatment regimes, including synchronous treatments. Agreed husbandry practices