Dataset Information

Neuronal gene expression in two generations of the marine parasitic worm, Cryptocotyle lingua.

ABSTRACT: Trematodes, or flukes, undergo intricate anatomical and behavioral transformations during their life cycle, yet the functional changes in their nervous system remain poorly understood. We investigated the molecular basis of nervous system function in Cryptocotyle lingua, a species of relevance for fisheries. Transcriptomic analysis revealed a streamlined molecular toolkit with the absence of key signaling pathways and ion channels. Notably, we observed the loss of nitric oxide synthase across the Platyhelminthes. Furthermore, we identified upregulated neuronal genes in dispersal larvae, including those involved in aminergic pathways, synaptic vesicle trafficking, TRPA channels, and surprisingly nitric oxide receptors. Using neuronal markers and in situ hybridization, we hypothesized their functional relevance to larval adaptations and host-finding strategies. Additionally, employing a behavior quantification toolkit, we assessed cercaria motility, facilitating further investigations into the behavior and physiology of parasitic flatworms. This study enhances our understanding of trematode neurobiology and provides insights for targeted antiparasitic strategies.

SUBMITTER: Tolstenkov O

PROVIDER: S-EPMC10728431 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Neuronal gene expression in two generations of the marine parasitic worm, Cryptocotyle lingua.

Tolstenkov Oleg O Chatzigeorgiou Marios M Gorbushin Alexander A

Communications biology 20231218 1

Trematodes, or flukes, undergo intricate anatomical and behavioral transformations during their life cycle, yet the functional changes in their nervous system remain poorly understood. We investigated the molecular basis of nervous system function in Cryptocotyle lingua, a species of relevance for fisheries. Transcriptomic analysis revealed a streamlined molecular toolkit with the absence of key signaling pathways and ion channels. Notably, we observed the loss of nitric oxide synthase across th ...[more]

PMID: 38110640

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Project description:BackgroundOne of the important challenges in microarray analysis is to take full advantage of previously accumulated data, both from one's own laboratory and from public repositories. Through a comparative analysis on a variety of datasets, a more comprehensive view of the underlying mechanism or structure can be obtained. However, as we discover in this work, continual changes in genomic sequence annotations and probe design criteria make it difficult to compare gene expression data even from different generations of the same microarray platform.ResultsWe first describe the extent of discordance between the results derived from two generations of Affymetrix oligonucleotide arrays, as revealed in cluster analysis and in identification of differentially expressed genes. We then propose a method for increasing comparability. The dataset we use consists of a set of 14 human muscle biopsy samples from patients with inflammatory myopathies that were hybridized on both HG-U95Av2 and HG-U133A human arrays. We find that the use of the probe set matching table for comparative analysis provided by Affymetrix produces better results than matching by UniGene or LocusLink identifiers but still remains inadequate. Rescaling of expression values for each gene across samples and data filtering by expression values enhance comparability but only for few specific analyses. As a generic method for improving comparability, we select a subset of probes with overlapping sequence segments in the two array types and recalculate expression values based only on the selected probes. We show that this filtering of probes significantly improves the comparability while retaining a sufficient number of probe sets for further analysis.ConclusionsCompatibility between high-density oligonucleotide arrays is significantly affected by probe-level sequence information. With a careful filtering of the probes based on their sequence overlaps, data from different generations of microarrays can be combined more effectively.

Project description:BackgroundIn the field of neuroscience microarray gene expression profiles on anatomically defined brain structures are being used increasingly to study both normal brain functions as well as pathological states. Fluorescent tracing techniques in brain tissue that identifies distinct neuronal populations can in combination with global gene expression profiling potentially increase the resolution and specificity of such studies to shed new light on neuronal functions at the cellular level.Methodology/principal findingsWe examine the microarray gene expression profiles of two distinct neuronal populations in the spinal cord of the neonatal rat, the principal motor neurons and specific interneurons involved in motor control. The gene expression profiles of the respective cell populations were obtained from amplified mRNA originating from 50-250 fluorescently identified and laser microdissected cells. In the data analysis we combine a new microarray normalization procedure with a conglomerate measure of significant differential gene expression. Using our methodology we find 32 genes to be more expressed in the interneurons compared to the motor neurons that all except one have not previously been associated with this neuronal population. As a validation of our method we find 17 genes to be more expressed in the motor neurons than in the interneurons and of these only one had not previously been described in this population.Conclusions/significanceWe provide an optimized experimental protocol that allows isolation of gene transcripts from fluorescent retrogradely labeled cell populations in fresh tissue, which can be used to generate amplified aRNA for microarray hybridization from as few as 50 laser microdissected cells. Using this optimized experimental protocol in combination with our microarray analysis methodology we find 49 differentially expressed genes between the motor neurons and the interneurons that reflect the functional differences between these two cell populations in generating and transmitting the motor output in the rodent spinal cord.

Dataset Information

Neuronal gene expression in two generations of the marine parasitic worm, Cryptocotyle lingua.

Publications

Neuronal gene expression in two generations of the marine parasitic worm, Cryptocotyle lingua.

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