Carlos Vedor (Inserm) has been focussing on developing a method for long read sequencing (using Nanopore technology) from single isolated cells. Despite all his effort, none of the strategies have managed the level of amplification desired. This outcome had been listed in the scientific risks we anticipated. The student is therefore now developing the mitigation strategy which will consist in using MARS-seq/SMART-seq protocols to maximize the sequencing coverage of isolated neurons.
David Kenigsberger (Weizmann) has made significant progress in dissecting the mechanisms underlying sexually dimorphic variability in nociceptive behavior in C. elegans. Building on previous work, the student successfully replicated behavioral assays showing that a subset of males responds to nociceptive stimulation in a hermaphrodite-like manner. Through artificial selection, David demonstrated that this phenotype can be transiently enriched across generations, supporting the hypothesis of a non-genetic, epigenetic mechanism.
Deborah Warrington (CERBM) During the first period, the student validated and extended analysis of embryonic scRNA-seq data, confirming pseudotime trajectories in vivo and constructing fluorescent reporters for downstream validation. She then shifted to the L1 stage: the larval stage preceding the Td initiation and analysed single-cell RNA-seq datasets of both Y and its non-reprogramming neighbour, the rectal cell B. This revealed surprising transcriptional transitions in B, likely reflecting maturation.
Claudio Lopez (UCBL/ULB). During the first year of the project we have performed several experiments exploring the potential effects of the parental pheromone perception in the dauer entry propensity of their F1 progeny. Preliminary results have suggested that there is an increase in the frequency of dauer entry in worms which parents have been exposed to high a concentration of ascarosides in comparison to the control. The project is progressing well despite the technical difficulty of the proposed methodology the student is steadily developing both his experimental and computational skills in the pursuit of his objectives.
Parichitran Ayyamperumal (UCBL) goal is to obtain a comprehensive understanding of the expression dynamics of potassium channel genes along the life of the worm, at the level of the entire nervous system. During the first period he first analysed stage-specific single-cell RNA sequencing datasets. At present, he generated translational and transcriptional reporter for 36 and 7 potassium channel genes, respectively. These reporters will be scored individually to determine gene expression (1) during larval development, (2) before, during and upon exit of the dauer stage, (3) in adult hermaphrodites vs. males. We will use different microscopy modalities depending for qualitative and quantitative assessments.
Yaren Canten (UU) has investigated UNC-33 levels and localization in various mutants that were previously reported to affect the protein. Also, she made knock-in strains for specific microtubule binding proteins to visualize UNC-33/MT dynamics in non-neuronal tissues. To further investigate the role of UNC-33 in non-neuronal tissues, she is currently developing tissue specific depletion assays. Since glia cells are important for proper neuon functioning she will address the importance of proper glia MT cytoskeleton organization on neuronal development and functioning using the above described tissue specific depletion tools.
Lisa Anderer (UU) is developing genetic tools and advanced microscopy approaches to study axonal transport at the endogenous level in the in vivo model C. elegans. Lisa is using the splitGFP and the FLP-FRT system to fluorescently label proteins of interest endogenously at the single-cell level. She has collected several existing strains and made several new ones labeling different neuronal cargos and motor proteins. She plans to perform proximity ligation experiments for which she will perform her secondment in the de Bono lab (Vienna) in early 2026. This work provides a foundation for studying how motor protein mediated cargo transport is regulated in neurons in vivo.
Anna Pellizzer (IST) investigated how two proteins of previously unknown function, MACO-1 and SUTU-7, co-translationally regulate biogenesis of polytopic membrane proteins at the endoplasmic reticulum (ER). She generated a bright reporter line for a client of the complex, a key step for further studies, and knockouts (KOs) for proteins involved in known ER-associated quality control mechanisms, including RDE-1 and NTL-3. She also investigated selected protein of unknown function that are conserved from C. elegans to humans. She used CRISPR to generate C. elegans knockouts for genes encoding the selected proteins, and assessed their behavioral phenotypes using computer-assisted video analyses (in particular through a secondment with Magnitude Bioscience).
Aditee Dandekar (ULB) aims are aim to characterize (1) whether and how PHAC-1 (PHACTRs) signaling control actin dynamics and how this modifies SV/DCV trafficking (2) the dynamic interactions of TOM-1 (STXBP5) with its presynaptic partner proteins (3) for this, we develop a single-molecule tracking approach for presynaptic proteins in the neuromuscular junction in-vivo. In the first reporting period she has progressed on all three objectives. During 2026, data collection in M. Harterink lab will map STXBP5 partitioning between vesicles, protein complexes, and membrane domains of presynapse for wild-type TOM-1::GFP. The phosphorylation of TOM-1 modulates its function, localization and synaptic strength.
