Data from: Effect of altered production and storage of dopamine on development and behavior in C. elegans
Data files
Aug 31, 2024 version files 41.59 GB
Abstract
Introduction: The nematode, Caenorhabditis elegans (C. elegans), is an advantageous model for studying developmental toxicology due to its well defined developmental stages and homology to humans. It has been established that across species, dopaminergic neurons are highly vulnerable to neurotoxicant exposure, resulting in developmental neuronal dysfunction and age-induced degeneration. C. elegans, with genetic perturbations in dopamine system proteins, can provide insight into the mechanisms of dopaminergic neurotoxicants. In this study, we present a comprehensive analysis on the effect of gene mutations in dopamine-related proteins on body size, development, and behavior in C. elegans.
Methods: We studied C. elegans that lack the ability to sequester dopamine (OK411) and that overproduce dopamine (UA57) and a novel strain (MBIA) generated by the genetic crossing of OK411 and UA57, which both lack the ability to sequester dopamine into vesicles and, additionally, endogenously overproduce dopamine. The MBIA strain was generated to address the hypothesis that an endogenous increase in the production of dopamine can rescue deficits caused by a lack of vesicular dopamine sequestration. These strains were analyzed for body size, developmental stage, reproduction, egg laying, motor behaviors, and neuronal health utilizing multiple methods.
Results: Our results further implicate proper dopamine synthesis and sequestration in the regulation of C. elegans body size, development through larval stages into gravid adulthood, and motor functioning. Furthermore, our analyses demonstrate that body size in terms of length is distinct from the developmental stage as fully developed gravid adult C. elegans with disruptions in the dopamine system have decreased body lengths. Thus, body size should not be used as a proxy for the developmental stage when designing experiments.
Discussion: Our results provide additional evidence that the dopamine system impacts the development, growth, and reproduction in C. elegans. Furthermore, our data suggest that endogenously increasing the production of dopamine mitigates deficits in C. elegans lacking the ability to package dopamine into synaptic vesicles. The novel strain, MBIA, and novel analyses of development and reproduction presented here can be utilized in developmental neurotoxicity experiments.
README: Effect of altered production and storage of dopamine on development and behavior in C. elegans
https://doi.org/10.5061/dryad.hhmgqnkpf
This dataset contains the raw data corresponding to the manuscript "Effect of altered production and storage of dopamine on development and behavior in C. elegans." The data presented here was in effort to understand and characterize the role of dopamine neurotransmission in C. elegans development utilizing genetic models. A novel strain was generated that lacks the protein cat-1, which is responsible for vesicular sequestration of dopamine, and over-expresses the gene cat-2, which is responsible for dopamine synthesis. Thus, this novel strain (MBIA) has compounded effects on the amount of cytosolic dopamine. We characterized this strain, the parent strains used to generate MBIA, and wild-type C. elegans to determine what role dopamine synthesis and sequestration has on body size, development through larval stages into gravid adulthood, reproduction, neuron health, and dopamine-mediated behaviors.
Description of the data and file structure
This dataset contains 5 compressed folders: 1. Behavior assay files, 2. CSV files, 3. Cleaned COPAS CSV files, 4. Brightfield and GFP images for body size and neuron area, and 5. MPP. Raw image and data files are included for each analysis. When appropriate, .csv files are provided.
Folder 1 contains subfolders for each developmental stage of analysis: L4 (larval stage 4), Day 1 (Day one adult), Day 2 (Day two adult), and Day 3 (Day three adult.) Within each subfolder labeled by developmental stage, there are four additional subfolders containing the dates on which the videos were recorded across four experimental replicates. Within each dated subfolder there are subfolders divided by strain: N2 (wild-type), UA57 (Cat-2 overexpression), Cat-1 (cat-1 null), and IA (MBIA, cat-2 overexpression and cat-1 null). Each strain subfolder contains multiple videos recorded in the format of sequenced .jpg files.
Folder 2 contains the .csv files corresponding to the statistics performed for the analysis.
Folder 3 contains the .csv files collected from the COPAS large particle biosorter arranged in subfolders corresponding to each cohort (labeled 1-6) on which the analysis was performed. Within each individual cohort subfolder there are individual .csv files corresponding to developmental stage (either L4 or Day 1 adult) and media formulation in which the c. elegans were grown (agar or liquid). Missing data code: NA
Folder 4 contains the raw image files compiled for microscopy analysis. There are brightfield images on which body length analysis was performed and GFP images on which neuron area analysis was performed.
Folder 5 contains all of the relevant files from the MPP+ exposure experiments found in the supplement of the manuscript organized in subfolders corresponding to the each cohort (labeled 1-5) on which the analysis was performed. Each individual cohort folder contains microscopy images corresponding to the developmental stage (L4 or day 1 adult) and doses of MPP+ treated (C025 = DMSO equivalent to 0.25uM dose, C075 = DMSO equivalent to 0.75uM dose, 25 = 0.25uM, 50 = 0.50um, 75 = 0.75um, NT = non-treated). Within Folder 5 is also the .csv file of the compiled eggs in utero data across the five cohorts of MPP+ experiments.