Ion channel profiling of the Lymnaea stagnalis ganglia via transcriptome analysis.

BACKGROUND: The pond snail Lymnaea stagnalis (L. stagnalis) has been widely used as a model organism in neurobiology, ecotoxicology, and parasitology due to the relative simplicity of its central nervous system (CNS). However, its usefulness is restricted by a limited availability of transcriptome data. While sequence information for the L. stagnalis CNS transcripts has been obtained from EST libraries and a de novo RNA-seq assembly, the quality of these assemblies is limited by a combination of low coverage of EST libraries, the fragmented nature of de novo assemblies, and lack of reference genome. RESULTS: In this study, taking advantage of the recent availability of a preliminary L. stagnalis genome, we generated an RNA-seq library from the adult L. stagnalis CNS, using a combination of genome-guided and de novo assembly programs to identify 17,832 protein-coding L. stagnalis transcripts. We combined our library with existing resources to produce a transcript set with greater sequence length, completeness, and diversity than previously available ones. Using our assembly and functional domain analysis, we profiled L. stagnalis CNS transcripts encoding ion channels and ionotropic receptors, which are key proteins for CNS function, and compared their sequences to other vertebrate and invertebrate model organisms. Interestingly, L. stagnalis transcripts encoding numerous putative Ca2+ channels showed the most sequence similarity to those of Mus musculus, Danio rerio, Xenopus tropicalis, Drosophila melanogaster, and Caenorhabditis elegans, suggesting that many calcium channel-related signaling pathways may be evolutionarily conserved. CONCLUSIONS: Our study provides the most thorough characterization to date of the L. stagnalis transcriptome and provides insights into differences between vertebrates and invertebrates in CNS transcript diversity, according to function and protein class. Furthermore, this study provides a complete characterization of the ion channels of Lymnaea stagnalis, opening new avenues for future research on fundamental neurobiological processes in this model system.

Factors Associated With Asthma Diagnosis Within Five Years of a Bronchiolitis Hospitalization: A Retrospective Cohort Study in a High Asthma Prevalence Population.

OBJECTIVES: Bronchiolitis, the leading cause of infant hospitalizations in the United States, is associated with increased risk of childhood asthma. We hypothesized that factors during a bronchiolitis hospitalization were associated with subsequent asthma. METHODS: This is a retrospective cohort study at an urban, tertiary-care children's hospital of infants <12 months old, hospitalized for bronchiolitis. The primary outcome measure was an asthma diagnosis, defined as a billing code for an asthma visit or a prescription for controller medication, within 5 years of discharge from the bronchiolitis hospitalization. RESULTS: There were 534 infants hospitalized for bronchiolitis, of which 294 (55.1%) were diagnosed with asthma, and 102 (19.1%) were hospitalized for asthma within 5 years of discharge. There was significant interaction between age and family history. In both models, female sex was protective (odds ratio [OR] 0.46). Age and race were only associated with asthma in infants without a family history of asthma: age (OR 1.19; 95% confidence interval 1.08-1.32) and race (OR 4.06; 95% confidence interval 1.56-10.58). Hospitalization length, ICU stay, albuterol treatments received, supplemental oxygen, respiratory support, highest respiratory rate, and respiratory syncytial virus infection were not associated with asthma diagnosis. CONCLUSIONS: More than 55% of infants hospitalized for bronchiolitis developed asthma within 5 years of discharge. Demographic and family history variables were independently associated with asthma. However, hospital-based variables during the bronchiolitis hospitalization were not independently associated with asthma. These results can direct further research and differentiate anticipatory guidance for infants with bronchiolitis at risk for asthma.

Pharmacotherapy in body dysmorphic disorder: relapse prevention and novel treatments.

Introduction: Body dysmorphic disorder is a debilitating disorder that often presents with significant delusionality, low insight, and both medical and psychiatric comorbidities, presenting a challenge for treatment and long-term management. Its typically chronic course requires that therapy be continued indefinitely, but only a few studies of long-term pharmacotherapeutic management exist. Areas covered: The authors discuss the current understanding of body dysmorphic disorder, focusing specifically on: epidemiology, clinical presentation, challenges in treatment, treatment options, and the importance of the further study of the long-term management of the disorder. Expert opinion: Serotonin reuptake inhibitors are the established drug of choice in patients with body dysmorphic disorder. Initial studies suggest that other agents such as augmentation antipsychotic medication may also be of use in combination with serotonin reuptake inhibitors, but there is a lack of studies comparing new treatments to serotonin reuptake inhibitors. Due to the chronic nature of body dysmorphic disorder, further research is needed to clarify the role of pharmacotherapy in long-term management and relapse prevention. Future studies should explore the long-term use of therapies and combinations of different therapeutics with the goal of effectively managing this debilitating, chronic condition.

MEN1 Tumor Suppressor Gene is Required for Long-term Memory Formation in an Aversive Operant Conditioning Model of Lymnaea stagnalis.

Activity-dependent transcription factors critically coordinate the gene expression program underlying memory formation. The tumor suppressor gene, MEN1, encodes a ubiquitously expressed transcription regulator required for synaptogenesis and synaptic plasticity in invertebrate and vertebrate central neurons. In this study, we investigated the role of MEN1 in long-term memory (LTM) formation in an aversive operant conditioning paradigm in the freshwater pond snail Lymnaea stagnalis (L. stagnalis). We demonstrated that LTM formation is associated with an increased expression of MEN1 coinciding with an up-regulation of creb1 gene expression. In vivo knockdown of MEN1 prevented LTM formation and conditioning-induced changes in neuronal activity in the identified pacemaker neuron RPeD1. Our findings suggest the involvement of a new pathway in LTM consolidation that requires MEN1-mediated gene regulation.

Dopamine-mediated calcium channel regulation in synaptic suppression in L. stagnalis interneurons.

D2 dopamine receptor-mediated suppression of synaptic transmission from interneurons plays a key role in neurobiological functions across species, ranging from respiration to memory formation. In this study, we investigated the mechanisms of D2 receptor-dependent suppression using soma-soma synapse between respiratory interneuron VD4 and LPeD1 in the mollusk Lymnaea stagnalis (L. stagnalis). We studied the effects of dopamine on voltage-dependent Ca2+ current and synaptic vesicle release from the VD4. We report that dopamine inhibits voltage-dependent Ca2+ current in the VD4 by both voltage-dependent and -independent mechanisms. Dopamine also suppresses synaptic vesicle release downstream of activity-dependent Ca2+ influx. Our study demonstrated that dopamine acts through D2 receptors to inhibit interneuron synaptic transmission through both voltage-dependent Ca2+ channel-dependent and -independent pathways. Taken together, these findings expand our understanding of dopamine function and fundamental mechanisms that shape the dynamics of neural circuit.

Inverse Relationship between Basal Pacemaker Neuron Activity and Aversive Long-Term Memory Formation in Lymnaea stagnalis.

Learning and memory formation are essential physiological functions. While quiescent neurons have long been the focus of investigations into the mechanisms of memory formation, there is increasing evidence that spontaneously active neurons also play key roles in this process and possess distinct rules of activity-dependent plasticity. In this study, we used a well-defined aversive learning model of aerial respiration in the mollusk Lymnaea stagnalis (L. stagnalis) to study the role of basal firing activity of the respiratory pacemaker neuron Right Pedal Dorsal 1 (RPeD1) as a determinant of aversive long-term memory (LTM) formation. We investigated the relationship between basal aerial respiration behavior and RPeD1 firing activity, and examined aversive LTM formation and neuronal plasticity in animals exhibiting different basal aerial respiration behavior. We report that animals with higher basal aerial respiration behavior exhibited early responses to operant conditioning and better aversive LTM formation. Early behavioral response to the conditioning procedure was associated with biphasic enhancements in the membrane potential, spontaneous firing activity and gain of firing response, with an early phase spanning the first 2 h after conditioning and a late phase that is observed at 24 h. Taken together, we provide the first evidence suggesting that lower neuronal activity at the time of learning may be correlated with better memory formation in spontaneously active neurons. Our findings provide new insights into the diversity of cellular rules of plasticity underlying memory formation.