Biology subjects

Dingle, L.

Publications and source records attributed to Dingle, L..

4 recordsLinked to original sources

bioRxiv2025

Neurotranscriptomic profiling of DWV-infected honey bee foragers with different cognitive abilities

Honey bees (Apis mellifera) provide important ecosystem services to both natural and human-managed environments, but are increasingly threatened by a variety of pathogens, the most common of which is deformed wing virus (DWV). DWV is known to replicate in the honey bee brain and has been documented as both improving and impairing olfactory learning and memory. We examined the transcriptomic response of the honey bee mushroom bodies--an area of the insect brain associated with higher cognitive functions--in bees with naturally occurring DWV infections who varied in their ability to perform an associative learning task. RNA-seq analysis detected increased expression of genes involved in the immune response, including important antimicrobial peptides (AMPs) such as hymenoptaecin, apidaecin, and abaecin, and the downreguation of lysozyme, PPO, and other genes associated with responses to a range of stressors. Additionally, gene ontology (GO) enrichment analysis revealed overrepresentation of key biological processes which form part of the immune response. We also noted significant differential expression of long non-coding RNAs (lncRNAs) presumed to be acting in a regulatory manner, and used these lncRNAs to construct gene regulatory networks (GRNs). Strikingly, in contrast to previous studies on bees with artificially-induced infections that have examined viral loads in the abdomen and non-specific areas of the brain, no correlation between DWV load in the mushroom bodies and cognitive function was noted. This highlights the complexity of host-pathogen interactions in honey bee neural tissues and the benefits of a spatially-refined approach to brain transcriptomics in naturally-occurring infections.

genomics↗

bioRxiv2023

Learning performance and GABAergic pathway link to deformed wing virus in the mushroom bodies of naturally infected honey bees

Viral infections can be detrimental to the foraging ability of the Western honey bee Apis mellifera. These include the deformed wing virus (DWV), which is the most common honey bee virus and has been proposed as a possible cause of learning and memory impairment. However, evidence for this phenomenon so far has come from artificially infected bees, while less is known about the implications of natural infections with the virus. Using the proboscis extension reflex (PER), we uncovered no significant association between a simple associative learning task and natural DWV loads. However, when assessed through a reversal associative learning assay, bees with higher DWV loads performed better in the reversal learning phase. DWV is able to replicate in the honey bee mushroom bodies, where the GABAergic signalling pathway has an antagonistic effect on associative learning but is crucial for reversal learning. Hence, we assessed the pattern of expression of several GABA-related genes in bees with different learning responses. Intriguingly, mushroom body expression of selected genes was positively correlated with DWV load, but only for bees with good reversal learning performance. We hypothesize that DWV might improve olfactory learning performance by enhancing the GABAergic inhibition of responses to unrewarded stimuli, which is consistent with the behavioural patterns that we observed. Our results suggest that previously reported DWV-driven learning deficits might be exclusive to acute, artificial infections and do not occur in naturally infected bees, stressing the importance of investigating more ecologically relevant scenarios when assessing host-parasite systems. Summary statementThis study describes a virus-associated increase in learning in honey bees and proposes a mechanism based on GABA to explain the interplay between infection and cognition in the insect brain.

animal behavior and cognition↗

bioRxiv2022

Novel Laser Capture Microdissection-Proteomic Analysis Identifies Spatially Distinct Extracellular Matrix Signatures in the Core and Infiltrating Edge of Human Glioblastoma.

BackgroundGlioblastoma is the most common and aggressive primary brain tumour in adults. Hallmarks of glioblastoma include its intra-tumoural heterogeneity and extensive invasion of the surrounding brain. Glioblastoma is known to remodel the extracellular matrix (ECM) of the brain, resulting in altered mechanical properties and the establishment of a tumour-promoting microenvironment. How changes in the expression and spatial distribution of ECM constituents within glioblastoma contribute to invasion and disease progression is still unclear. MethodsHere we report on a protocol for laser-capture microdissection coupled with mass spectrometry (LCM-proteomics) that allowed a spatially resolved and unbiased analysis of the regional ECM proteome (matrisome) in formalin-fixed and paraffin-embedded (FFPE) samples of human glioblastoma. We investigated five molecularly characterised hemispheric adult glioblastomas where the brain/tumour interface and tumour epicentre were represented in the surgical specimens and snap-frozen tissue was available. LCM-proteomic analysis was validated with immunohistochemistry. ResultsLCM-proteomics identified 53 matrisome proteins in FFPE tissue, demonstrating comparable performance with conventional analysis of snap-frozen tissue. The analysis revealed distinct matrisome components in the brain/tumour interface versus the tumour epicentre. Guided by data from LCM-proteomic analysis, immunostaining for tenascin-R confirmed greater staining in the brain/tumour interface, whilst expression of fibronectin was higher in the tumour epicentre. ConclusionThe protocol described in this work allowed for accurate, spatially resolved analysis of ECM in FFPE tissues, with performance comparable to analysis of snap-frozen tissue. While the focus for this work was on the regional ECM composition of glioblastoma, we found that the LCM-proteomics protocol is also applicable to the study of the wider proteome, and represents a valuable tool for investigating tumour/tissue heterogeneity. This protocol opens the possibility to apply LCM-proteomics to retrospective studies with the advantage of accessing clinical history and follow-up information, providing a valuable resource for translational research in glioblastoma.

cancer biology↗

bioRxiv2021

The IPF fibroblastic focus is an active collagen biosynthesis factory embedded in a distinct extracellular matrix

BackgroundThe Fibroblastic Focus (FF) is the signature lesion of Idiopathic Pulmonary Fibrosis (IPF) where myofibroblasts accumulate and extracellular matrix (ECM) is produced. However, the molecular composition and function of the FF and surrounding tissue remain undefined. MethodsUtilizing laser capture microdissection coupled mass spectrometry (LCM-MS), we interrogated the FF, adjacent mature scar, and adjacent alveoli in 6 IPF specimens plus 6 non-fibrotic alveolar specimens as controls. The data were subject to qualitative and quantitative analysis, and validation by immunohistochemistry. ResultsWe found that the protein signature of IPF alveoli is defined by immune deregulation as the strongest category. The IPF mature scar was classified as end-stage fibrosis whereas the FF contained an overabundance of a distinctive ECM compared to non-fibrotic control. ConclusionSpatial proteomics demonstrated distinct protein compositions in the histologically defined regions of IPF tissue. These data revealed that the FF is the main site of collagen biosynthesis and that the alveoli adjacent to the FF are abnormal. This new and essential information will inform future mechanistic studies on mechanisms of IPF progression.

pathology↗